Preparation of terpenoid compounds

Abstract

The present invention provides a method for the preparation of a compound of the formula (II-1), (II-2), (II-3) and / or (II-4). Said compound may be used as a perfumery, flavor or aroma ingredient, and / or as a precursor thereof. Included in this invention is an in vivo process for the preparation of said compound in recombinant cells. The present invention also provides recombinant cells which may be used in said method. Novel compounds are also part of the invention.

Description

[0001] PREPARATION OF TERPENOID COMPOUNDS

[0002] Technical field

[0003] The present invention relates to the field of biosynthesis. More specifically, it concerns a method for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4) starting from compound of the formula (I). The compounds of the formula (11-1-1), (11-1-1 a), (11-1-2), (ll-1-2a), (II-1-3), (ll-1-3a), (11-1-4), (11-1 -4a), (11-1-5), (ll-1-5a), (ll-4-6a), (II-Aa), (11-1 b), (ll-1c) and (ll-1d) are also part of the invention. Said compounds may be used as a perfumery, flavor or aroma ingredient and / or a precursor thereof.

[0004] Background

[0005] In the perfumery, flavor or aroma industry, there is a constant need to provide new compounds and / or new methods for the preparation thereof. Key amongst such compounds are terpene compounds which are naturally found in most organisms (microorganisms, animals and plants). These compounds are made up of five carbon units called isoprene units and are classified by the number of these units present in their structure. Thus monoterpenes, sesquiterpenes and diterpenes are terpenes containing 10, 15 and 20 carbon atoms respectively. Sesquiterpenes, for example, are widely found in the plant kingdom. Many sesquiterpene molecules are known for their flavor and fragrance properties and their cosmetic, medicinal and antimicrobial effects. Numerous sesquiterpene hydrocarbons and sesquiterpenoids have been identified. Diterpenes are formed from four isoprene units, 20 carbons, and are secondary metabolites identified in plants and fungi. Although most diterpenes are non-volatile and some are present in essential oils.

[0006] Biosynthetic production of terpene compounds usually involves enzymes called terpene synthases. These enzymes convert an acyclic terpene precursor in one or more terpene products. In particular, diterpene synthases produce diterpenes by cyclization of the precursor geranylgeranyl diphosphate (GGPP). The cyclization of GGPP often requires two enzyme polypeptides, a type I and a type II diterpene synthase working in combination in two successive enzymatic reactions. The type II diterpene synthases catalyze a cyclization / rearrangement of GGPP initiated by the protonation of the terminal double bond of GGPP leading to a cyclic diterpene diphosphate intermediate. This intermediate is then further converted by a type I diterpene synthase catalyzing an ionization-initiated cyclization.

[0007] Despite the improvements that have been made in this field, there nevertheless remains a need for new methods of producing terpene compounds which are building blocks for the preparation of highly valuable perfumery, flavor or aroma ingredients.

[0008] This problem is addressed by the present invention which provides a novel method comprising an enzymatic step for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4) starting from compound of the formula (I). Summary

[0009] The present invention is based on the surprising finding of the inventors that meroterpenoid cyclases can be used in the cyclisation of linear terpenoid compounds to produce a compound of formula (11-1), (II-2), (II-3) and / or (II-4). Accordingly, the invention provides a new method for the preparation of a compound of the formula (11-1), (II-2), (II-3) and / or (II-4). This aspect of the invention and the compound of the formula (11-1), (II-2), (II-3) and / or (II-4) enable new routes toward the preparation of perfumery, flavor or aroma compounds.

[0010] A first aspect of the invention provides a method for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4),

[0011]

[0012]

[0013] and / or

[0014] in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises more than one--OH group,

[0015] a compound having at least one-_O-- group,

[0016] which is formally obtained by an intramolecular etherification or esterification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification or esterification, two--OH groups form, under cleavage of a molecule of water, a cyclic —O— group;

[0017] and / or

[0018] in the case where the compound of the formula (11-1) or (I I-2) or (I I-3) or (I I-4) comprises at least two--OH groups and at least one carbonyl group,

[0019] a compound having at least one-_O-- group,

[0020] which is formally obtained by ketalization of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the ketalization a carbonyl group and two--OH groups form, under cleavage of water, a cyclic-_O-- group;

[0021] and / or

[0022] in the case where the compound of the formula (11-1) or (I I-2) or (I I-3) or (I I-4) comprises at least one--OH group and at least one carbonyl group,

[0023] a compound having at least one-_O-- group,

[0024] which is formally obtained by etherification of compound of the formula (11-1) or (II-2) or (II-3) or (II-4), wherein in the etherification an--OH group a carbonyl group having undergone enolization form, under cleavage of water, a cyclic-_O-- group;

[0025] or a derivative thereof,

[0026] with the proviso that said compound is not

[0027]

[0028] wherein

[0029] A represents

[0030]

[0031] A' represents H, or

[0032] A' represents

[0033]

[0034] R represents

[0035]

[0036] H, wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;

[0037] wherein R2= H or an alcohol protecting group, particularly

[0038]

[0039] '' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and

[0040] m = 0 or 1 or 2;

[0041] n = 0 or 1 or 2;

[0042] with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;

[0043] z = 1 or 2;

[0044] R0independently form each other represents H or CH3or CH2OH;

[0045] wherein

[0046] any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration; and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond;

[0047] characterized in that the method comprises:

[0048] (a) contacting a compound of the formula (I)

[0049]

[0050] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-1), (II-2), (II-3) and / or (II-4).

[0051] An embodiment of the invention is wherein the compound of having at least one — O— group is selected from the group consisting of the compounds of the formula

[0052]

[0053] (ll-E), and

[0054]

[0055] An embodiment of the invention is wherein the compound of the formula (I) is selected from the group consisting of the compounds of the formula

[0056]

[0057] (l-r), and

[0058] (l-s).

[0059]

[0060] An embodiment of the invention is wherein the terpene cyclase is a meroterpenoid cyclase enzyme; preferably, a bacterial membrane-integrated meroterpenoid cyclase enzyme.

[0061] A further embodiment of the invention is wherein the method is an in vivo or a bioconversion process.

[0062] A further embodiment of the invention is wherein said method is performed in a recombinant cell capable of functionally expressing the meroterpenoid cyclase enzyme as defined herein above.

[0063] A futher aspect of the invention provides a compound selected from the group consisting of the compounds of the formula (11-1 a), (11-1 b), (11-1 c), and (11-1 d); preferably, a compound selected from the group consisting ofthe compounds of the formula (11-1-1), (11-1 -1 a), (11-1-2), (II-1-2a), (II-1-3), (11-1 -3a), (II-1-4), (II-1-4a), (11-1-5), (ll-1-5a), (ll-4-6a), (II-Aa), (11-1 b), (11-1 c) and (ll-1d).

[0064] A further aspect ofthe invention provides a recombinant cell comprising, capable of producing or producing a compound of the formula (II-1), (II-2), (II-3) and / or (II-4); preferably, a compound of the formula (11-1 a), (11-1 b), (11-1 c), and / or (ll-1d); more preferably, a compound of the formula (11-1-1), (11-1-1a), (11-1-2), (ll-1-2a), (II-1-3), (ll-1-3a), (11-1-4), (II-1-4a), (11-1-5), (ll-1-5a), (ll-4-6a), (II-Aa), (11-1 b), (ll-1c) and / or (11-1 d).

[0065] A further aspect of the invention provides the use of the compound of the invention as a perfumery, flavor or aroma ingredient, or as a precursor thereof.

[0066] A further aspect ofthe invention provides the use of a meroterpenoid cyclase enzyme to produce the compound ofthe invention. of the

[0067] Figure 1. (a) Biosynthetic pathway of E,E,E-geranylgeraniol (I-c) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP), (b) Biosynthetic pathway of Z,E-farnesol (I-d) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP), (c) Biosynthetic pathway of E,E-farnesol (I-e), E,E-farnesoic acid (I-k) and E,E-methyl-farnesoate (I-m) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP), (d) Biosynthetic pathway of Z,Z-farnesol (I-f) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP), (e) Biosynthetic pathway of (R)-nerolidol (I-oa) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP), (f) Biosynthetic pathway of (S)-nerolidol (I-ob) from isopentenyl-diphosphate (IPP) and dimethylallyl-diphosphate (DMAPP). Figure 2. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (I-a) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_108066363.1 (SEQ ID NO: 38).

[0068] Figure 3. Electron impact (EI) mass spectrum (MS) of the compound of the formula (II-2-17a).

[0069] Figure 4. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (I-b) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase (a) RLD81128.1 (SEQ ID NO: 20), (b) WP_194974368.1 (SEQ ID NO: 46), (c) HBB88633.1 (SEQ ID NO: 2).

[0070] Figure 5. Electron impact (EI) mass spectrum (MS) of (a) the compounds of the formula (II-2-7a), (II-4-3a), (II-1-3a) and (II-2-9), (b) the compounds of the formula (II-2-8a) and (II-3-2a).

[0071] Figure 6. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the proteins PvCPS-del623 (SEQ ID NO: 158) and MtRv3376 (SEQ ID NO: 159) for the production of the compound of the formula (I-c) and the bacterial membrane-integrated meroterpenoid cyclase (a) WP_057983506.1 (SEQ ID NO: 34), (b) WP_206728286.1 (SEQ ID NO: 134), (c) MCU1478099.1 (SEQ ID NO: 16).

[0072] Figure 7. Electron impact (EI) mass spectrum (MS) of the compounds of the formula (II-4-4a), (II-2-10a) and (II-3-3a).

[0073] Figure 8. GC-MS analysis of cyclic compounds produced using E. coli (DP1205 strain with FPP synthase deletion) cells expressing the proteins AgGPPS-delta 86 (SEQ ID NO: 152), ezFPS (SEQ ID NO: 153) and TalCeTpp (SEQ ID NO: 151) for the production of the compound of the formula (I-d) and the bacterial membrane-integrated meroterpenoid cyclase RLD81128.1 (SEQ ID NO: 20).

[0074] Figure 9. Electron impact (EI) mass spectrum (MS) of (a) the compounds of the formula (II-Aa), (II-2-4a), (II-4-1b) and (II-1-2a), (b) the compounds of the formula (II-2-5) and (II-4-2b).

[0075] Figure 10. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the protein TalVeTPP (SEQ ID NO: 150) for the production of the compound of the formula (I-e) and the bacterial membrane-integrated meroterpenoid cyclase (a) WP_194974368.1 (SEQ ID NO: 46), (b) RLD81128.1 (SEQ ID NO: 20).

[0076] Figure 11. Electron impact (EI) mass spectrum (MS) of (a) the compounds of the formula (II-2-2a), (II-2-1), (II-2-3a) and (II-1-1a), (b) the compounds of the formula (II-3-1a), (II-4-2a) and (II-4-1a).

[0077] Figure 12. GC-MS analysis of cyclic compounds produced using E. coli DP1205 (DP1205 strain with FPP synthase deletion) cells expressing the proteins B8XA40.1 (SEQ ID NO: 154) and TalCeTpp (SEQ ID NO: 151) for the production of the compound of the formula (I-f) and the bacterial membrane-integrated meroterpenoid cyclase RLD81128.1 (SEQ ID NO: 20).

[0078] Figure 13. Electron impact (EI) mass spectrum (MS) of the compound of the formula (II-2-4b).

[0079] Figure 14. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of formula (I-g) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_229232892.1 (SEQ ID NO: 55).

[0080] Figure 15. Electron impact (EI) mass spectrum (MS) of the compound of the formula (II-Ba).

[0081] Figure 16. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of formula (I-h) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_257961400.1 (SEQ ID NO: 63).

[0082] Figure 17. Electron impact (EI) mass spectrum (MS) of the compounds of the formula (II-Ca) and (II-4-5a).

[0083] Figure 18. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of formula (I-i) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase HBB88633.1 (SEQ ID NO: 2).

[0084] Figure 19. Electron impact (EI) mass spectrum (MS) of the compounds of formula (II-Cb), (II-4-5b) and (II-4-6a).

[0085] Figure 20. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the proteins TalCeTPP (SEQ ID NO: 151), CdGeoA (SEQ ID NO: 155) and SCH25-ALDH3 (SEQ ID NO: 156) for the production of the compound of the formula (I-k) and the bacterial membrane-integrated meroterpenoid cyclase (a) RLD81128.1 (SEQ ID NO: 20), (b) WP_194974368.1 (SEQ ID NO: 46). Figure 21. Electron impact (EI) mass spectrum (MS) of the compounds of the formula (II-2-12a), (II-1-4a), (II-2-14a) and (II-2-13).

[0086] Figure 22. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of formula (I-l) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_139559286.1 (SEQ ID NO: 129).

[0087] Figure 23. Electron impact (EI) mass spectrum (MS) of the compound of the formula (II-Ea). Figure 24. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the proteins TalCeTPP (SEQ ID NO: 151), CdGeoA (SEQ ID NO: 155), SCH25-ALDH3 (SEQ ID NO: 156) and NP_609793.2 (SEQ ID NO: 157) for the production of the compound of the formula (l-m) and the bacterial membrane-integrated meroterpenoid cyclase (a) WP_194974368.1 (SEQ ID NO: 46), (b) RLD81128.1 (SEQ ID NO: 20).

[0088] Figure 25. Electron impact (El) mass spectrum (MS) of the compounds of the formula (II-2-15a), (II-1-5a) and (II-2-16).

[0089] Figure 26. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-n) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_229232892.1 (SEQ ID NO: 55).

[0090] Figure 27. Electron impact (El) mass spectrum (MS) of the compound of the formula (II-3-5a).

[0091] Figure 28. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the protein SCH54Tps05901 (SEQ ID NO: 161) for the production of the compound of the formula (l-oa) and the bacterial membrane-integrated meroterpenoid cyclase RLD81128.1 (SEQ ID NO: 20).

[0092] Figure 29. Electron impact (El) mass spectrum (MS) of the compound of formula (ll-Fa).

[0093] Figure 30. GC-MS analysis of cyclic compounds produced using E. coli DP1205 cells expressing the protein ABR24417.1 (SEQ ID NO: 160) for the production of the compound of the formula (l-ob) and the bacterial membrane-integrated meroterpenoid cyclase RLD81128.1 (SEQ ID NO: 20).

[0094] Figure 31. Electron impact (El) mass spectrum (MS) of the compound of formula (II-Fb).

[0095] Figure 32. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-p) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_206728286.1 (SEQ ID NO: 134).

[0096] Figure 33. Electron impact (El) mass spectrum (MS) of the compounds of the formula (II-2-11a) and (II-3-4a).

[0097] Figure 34. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-q) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase (a) WP_251045335.1 (SEQ ID NO: 62), (b) WP_108030424.1 (SEQ ID NO: 37).

[0098] Figure 35. Electron impact (El) mass spectrum (MS) of the compounds of the formula (II-3-6a) and (II-4-7a).

[0099] Figure 36. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-f) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase A0A2M7NI22 (SEQ ID NO: 162). Figure 37. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-r) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WP_318017018.1 (SEQ ID NO: 73).

[0100] Figure 38. Electron impact (El) mass spectrum (MS) of the compounds of the formula (II-3-7a).

[0101] Figure 39. Biosynthetic pathway of compound of the formula (II-3-6a), coumpound of the formula (II-3-8) and compound of the formula (l-s) from E, E-Farnesyl diphosphate (E. E-FPP).

[0102] Figure 40. GC-MS analysis of compounds produced using E. coli DP1205 cells expressing (A) the geranylgeraniol diphosphate synthase CrtE (SEQ ID NO: 223) for the production of the compound of the formula (l-s), the bacterial membrane-integrated meroterpenoid cyclase WP_234754442.1 (SEQ ID NO: 79) and the phosphatase PgpB (SEQ ID NO: 224); (B) the geranylgeraniol diphosphate synthase CrtE (SEQ ID NO: 223), the bacterial membrane-integrated meroterpenoid cyclase WP_234754442.1 (SEQ ID NO: 79) and the sclareol synthase SaSCS (Seq ID NO: 225); or (C) the geranylgeraniol diphosphate synthase CrtE (SEQ ID NO: 223) and the sclareol synthase SaSCS (Seq ID NO: 225).

[0103] Figure 41. GC-MS analysis of cyclic compounds produced by the bioconversion of the compound of the formula (l-j) with E. coli C43(DE3) cells expressing the bacterial membrane-integrated meroterpenoid cyclase WSW93534.1 (SEQ ID NO: 74).

[0104] Figure 42. Electron impact (El) mass spectrum (MS) of the compounds of the formula (II-Da). Description of the Sequences

[0105] SEQ Description Name Origin

[0106] ID

[0107] 1 Meroterpenoid cyclase ASN53966.1 Sinomonas sp. R1AF57

[0108] 2 Meroterpenoid cyclase HBB88633.1 Blastocatellia bacterium

[0109] 3 Meroterpenoid cyclase MBD5477212.1 Lachnospiraceae bacterium

[0110] 4 Meroterpenoid cyclase MBI3240951.1 Chloroflexi bacterium

[0111] 5 Meroterpenoid cyclase MBI4522454.1 Deltaproteobacteria bacterium

[0112] 6 Meroterpenoid cyclase MBK6327782.1 Chloroflexi bacterium

[0113] 7 Meroterpenoid cyclase MBK6729680.1 Bacteroidetes bacterium

[0114] 8 Meroterpenoid cyclase MBK6918764.1 Deltaproteobacteria bacterium

[0115] 9 Meroterpenoid cyclase MBQ6784883.1 Clostridia bacterium

[0116] 10 Meroterpenoid cyclase MBQ8925419.1 Clostridia bacterium

[0117] 11 Meroterpenoid cyclase MBS1595609.1 Bacteroidota bacterium

[0118] 12 Meroterpenoid cyclase MBS1618437.1 Bacteroidota bacterium

[0119] 13 Meroterpenoid cyclase MBS1684121.1 Bacteroidetes bacterium

[0120] 14 Meroterpenoid cyclase MCA9899365.1 Anaerolineales bacterium

[0121] 15 Meroterpenoid cyclase MCB0700607.1 Chitinophagaceae bacterium

[0122] 16 Meroterpenoid cyclase MCU1478099.1 Subtercola sp.

[0123] 17 Meroterpenoid cyclase MCV67806.1 Listeria monocytogenes

[0124] 18 Meroterpenoid cyclase PKL36047.1 Spirochaetae bacterium HGW-Spirochaetae-1 19 Meroterpenoid cyclase RAN69340.1 Bacillus sp. SRB_336

[0125] 20 Meroterpenoid cyclase RLD81128.1 Bacteroidota bacterium

[0126] 21 Meroterpenoid cyclase TAN15888.1 Chitinophagaceae bacterium

[0127] 22 Meroterpenoid cyclase TLU50760.1 Chlorobium sp.

[0128] 23 Meroterpenoid cyclase TMI78539.1 Bacteroidota bacterium

[0129]

[0130] Meroterpenoid cyclase UXX94839.1 Streptomyces sp. Meroterpenoid cyclase WP_007916058.1 Ktedonobacter racemifer Meroterpenoid cyclase WP_019072542.1 Streptomyces hokutonensis Meroterpenoid cyclase WP_025162987.1 Paraclostridium bifermentans Meroterpenoid cyclase WP_028277553.1 Arthrobacter sp. 13 Meroterpenoid cyclase WP_033281172.1 Streptomyces sp. NRRL F-525 Meroterpenoid cyclase WP_036124658.1 Lysinibacillus

[0131] Meroterpenoid cyclase WP_043407626.1 Archangium violaceum Meroterpenoid cyclase WP_053678879.1 Streptomyces sp. WM4235 Meroterpenoid cyclase WP_057310159.1 Paenibacillus sp. Soil766 Meroterpenoid cyclase WP_057983506.1 Solibacillus cecembensis Meroterpenoid cyclase WP_077952889.1 Listeria monocytogenes Meroterpenoid cyclase WP_092727888.1 Romboutsia lituseburensis Meroterpenoid cyclase WP_108030424.1 Lysinibacillus tabacifolii Meroterpenoid cyclase WP_108066363.1 Vitiosangium sp. GDMCC 1.1324 Meroterpenoid cyclase WP_114779196.1 Botryobacter ruber Meroterpenoid cyclase WP_123134027.1 Rufibacter immobilis Meroterpenoid cyclase WP_133058268.1 Butyricicoccus porcorum Meroterpenoid cyclase WP_149098278.1 Rufibacter glacialis Meroterpenoid cyclase WP_163288115.1 Diaminobutyricibactertongyongensis Meroterpenoid cyclase WP_183976775.1 Runella defluvii Meroterpenoid cyclase WP_190963420.1 Desmonostoc muscorum Meroterpenoid cyclase WP_194974368.1 Aquiflexum lacus Meroterpenoid cyclase WP_200689660.1 Streptomyces sp.

[0132] Meroterpenoid cyclase WP_200714303.1 Streptomyces sp.

[0133] Meroterpenoid cyclase WP_205832565.1 Bifidobacterium sp. DSM 109959

[0134]

[0135] Meroterpenoid cyclase WP_206725628.1 Pyxidicoccus parkwaysis Meroterpenoid cyclase WP_207430198.1 Sabulibacter ruber Meroterpenoid cyclase WP_210464836.1 Rufibacter sp. SYSU D00344 Meroterpenoid cyclase WP_219871081.1 Paenibacillus oenotherae Meroterpenoid cyclase WP_220206969.1 Reticulibacter mediterranei Meroterpenoid cyclase WP_229232892.1 Corynebacterium cyclohexanicum Meroterpenoid cyclase WP_234333798.1 Streptomyces lavendulae Meroterpenoid cyclase WP_234351487.1 Streptomyces sp. XY413 Meroterpenoid cyclase WP_241253585.1 Streptomyces sp. W1SF4 Meroterpenoid cyclase WP_241642485.1 Pontibacter sp. E15-1 Meroterpenoid cyclase WP_242598968.1 Arthrobacter bambusae Meroterpenoid cyclase WP_248843460.1 Streptomyces virginiae Meroterpenoid cyclase WP_251045335.1 Arthrobacter sp. ISL-5 Meroterpenoid cyclase WP_257961400.1 Lysinibacillus capsici Meroterpenoid cyclase WP_261993638.1 Streptomyces sp. 3211.6 Meroterpenoid cyclase WP_262429210.1 Paratissierella segnis Meroterpenoid cyclase WP_266771239.1 Streptomyces sp.

[0136] Meroterpenoid cyclase WP_266817465.1 Streptomyces sp. NBC_00160 Meroterpenoid cyclase WP_266907787.1 Streptomyces sp. NBC_00086 Meroterpenoid cyclase WP_267502276.1 Streptomyces flavotricini Meroterpenoid cyclase WP_280879230.1 Streptomyces pseudovenezuelae Meroterpenoid cyclase WP_317769678.1 Streptomyces prunicolor Meroterpenoid cyclase WP_317880339.1 Streptomyces sp.

[0137] Meroterpenoid cyclase WP_318017018.1 Streptomyces sp.

[0138] Meroterpenoid cyclase WSW93534.1 Streptomyces sp.

[0139] Meroterpenoid cyclase WSX12386.1 Streptomyces sp.

[0140]

[0141] Meroterpenoid cyclase WSY54005.1 Streptomyces sp.

[0142] Meroterpenoid cyclase WTE43000.1 Streptomyces sp.

[0143] Meroterpenoid cyclase WTK72062.1 Streptomyces sp.

[0144] Meroterpenoid cyclase WP_234754442.1 Arthrobacter ramosus Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with S9M _S9M

[0145] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with A118M _A118M

[0146] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with A118P _A118P

[0147] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with C-terminal _d210 truncation of 21 amino acids Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with C-terminal _d217 truncation of 14 amino acids Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with F127W _F127W

[0148] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with F127Y _F127Y

[0149] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with N51 M _N51M

[0150] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with N51 Q _N51Q

[0151] Meroterpenoid cyclase WP_033281172.1 Mutant of SEQ ID NO: 29 with Q59M _Q59M

[0152] Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with N-terminal _dll truncation of 11 amino acids Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with N-terminal _d21 truncation of 21 amino acids Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with F157M _F157M

[0153] Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with F188M

[0154] _F188M

[0155]

[0156] Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with F27M _F27M

[0157] Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with G68D _G68D

[0158] Meroterpenoid cyclase WP_194974368.1 Mutant of SEQ ID NO: 46 with I53A _I53A

[0159] Meroterpenoid cyclase MBK8486394.1 Bacteroidota bacterium Meroterpenoid cyclase MBN3583158.1 Algoriphagus aestuarii Meroterpenoid cyclase MBU3664136.1 Bacteroidetes bacterium Meroterpenoid cyclase MBU3680180.1 Candidatus Kapabacteria.

[0160] Meroterpenoid cyclase WP_008199852.1 Algoriphagus machipongonensis Meroterpenoid cyclase EAE8345841.1 Listeria monocytogenes Meroterpenoid cyclase EHB56414.1 Mycolicibacterium rhodesiae JS60 Meroterpenoid cyclase HCZ38171.1 Brochothrix thermosphacta Meroterpenoid cyclase KMY33005.1 Lysinibacillus xylanilyticus Meroterpenoid cyclase KRE49200.1 Paenibacillus sp. Soil724D2 Meroterpenoid cyclase KRN43033.1 Fructilactobacillus fructivorans Meroterpenoid cyclase MBB3067326.1 Paenibacillus baekrokdamisoli Meroterpenoid cyclase MBC8085864.1 Phycisphaerae bacterium Meroterpenoid cyclase MBE3559230.1 Ktedonobacteraceae bacterium Meroterpenoid cyclase MBQ0136375.1 Oscillospiraceae bacterium Meroterpenoid cyclase MBQ1297511.1 Clostridiales bacterium Meroterpenoid cyclase MBQ2567061.1 Clostridia bacterium Meroterpenoid cyclase MBQ6526281.1 Clostridia bacterium Meroterpenoid cyclase MCB1397132.1 Rhodobacteraceae bacterium Meroterpenoid cyclase MCP2168610.1 Goodfellowiella coeruleoviolacea Meroterpenoid cyclase MCU1595540.1 Frankiales bacterium Meroterpenoid cyclase MVF22416.1 Methylocaldum sp. BRCS4

[0161]

[0162] Meroterpenoid cyclase NBR85593.1 Verrucomicrobia bacterium

[0163] Meroterpenoid cyclase NUT99169.1 Saccharothrix sp.

[0164] Meroterpenoid cyclase OFW07828.1 Acidobacteria bacterium

[0165] Meroterpenoid cyclase RAJ69904.1 Streptomyces sp. Amel2xB2 Meroterpenoid cyclase RDD90010.1 Streptomyces parvulus

[0166] Meroterpenoid cyclase SCI03284.1 Clostridium sp.

[0167] Meroterpenoid cyclase WP_055470795.1 Streptomyces pathocidini

[0168] Meroterpenoid cyclase WP_095978564.1 Melittangium boletus

[0169] Meroterpenoid cyclase WP_100897627.1 Nostoc flagelliforme

[0170] Meroterpenoid cyclase WP_111316424.1 Algoriphagus chordae

[0171] Meroterpenoid cyclase WP_139559286.1 Methylotetracoccus oryzae

[0172] Meroterpenoid cyclase WP_192347271.1 Algoriphagus sp. Y33

[0173] Meroterpenoid cyclase WP_196434265.1 Methylomonas sp. LL1

[0174] Meroterpenoid cyclase WP_200803587.1 Hespellia stercorisuis

[0175] Meroterpenoid cyclase WP_203896506.1 Actinoplanes xinjiangensis

[0176] Meroterpenoid cyclase WP_206728286.1 Pyxidicoccus parkwaysis

[0177] Meroterpenoid cyclase WP_235159650.1 Dyadobacter sp. CY351

[0178] Meroterpenoid cyclase WP_235179489.1 Dyadobacter chenhuakuii

[0179] Meroterpenoid cyclase WP_244504854.1 Lysinibacillus

[0180] Meroterpenoid cyclase WP_264061028.1 Mycobacterium montefiorense Meroterpenoid cyclase WP_270878872.1 Paenibacillus aestuarii

[0181] Meroterpenoid cyclase WP_271813414.1 Clostridium beijerinckii

[0182] Meroterpenoid cyclase WP_294903194.1 Lacibacter sp.

[0183] Squalene cyclase AAcSHC_M132R_ Mutant of squalene cyclase from Alicyclobacillus W169G_I432T_ acidocaldarius with M132R, W169G, I432T, G600M

[0184] G600M

[0185] Squalene cyclase ZmSHC_F503A_ Mutant of squalene cyclase from Zymomonas G667M mobilis with F503A, G667M

[0186]

[0187] Squalene cyclase BmeSHC_G595M Mutant of squalene cyclase from Priestia megaterium with G595M

[0188] Squalene cyclase AAcSHC_F437A_ Mutant of squalene cyclase from Alicyclobacillus G600M acidocaldarius with F437A, G600M Squalene cyclase GmoSHC_E46Q Mutant of squalene cyclase from Gluconobacter morbifer with E46Q

[0189] Squalene cyclase GmoSHC_Q54E_ Mutant of squalene cyclase from Gluconobacter M184I_V45L_T32 morbifer with Q54E, M184I, V45L, T326S, F624Y

[0190] 6S_F624Y

[0191] Squalene cyclase AAcSHC_F129A_ Mutant of squalene cyclase from Alicyclobacillus F437L_G600M acidocaldarius with F129A, F437L, G600M T rans-Beta-Farnesene AAX39387.1 Artemisia annua

[0192] synthase (AaBFS)

[0193] Phosphatase TalVeTPP Talaromyces verruculosus

[0194] Phosphatase TalCeTpp Talaromyces cellulolyticus

[0195] GPP synthase AgGPPS-delta 86 GPP synthase from Abies grandis with N- terminal truncation of 86 amino acids

[0196] Cis-IPP transferase ezFPS Mycobactrium tuberculosis

[0197] Z, Z-farnesol synthase B8XA40.1 Solanum habrochaites

[0198] Alcohol CdGeoA Castellaniella defragrans

[0199] dehydrogenase

[0200] Aldehyde SCH25-ALDH3 Cryptococcus laurentii

[0201] dehydrogenase

[0202] Farnesoic acid O- NP_609793.2 Drosophila melanogaster

[0203] methyl transferase

[0204] GGPP synthase PvCPS-del623 GGPP synthase from Talaromyces verruculosus with N-terminal truncation of 623 amino acids GGPP phosphatase MtRv3376 Mycobacterium tuberculosis

[0205] S-Nerolidol Synthase ABR24417.1 Antirrhinum majus

[0206] R-Nerolidol Synthase SCH54Tps05901 Trichoderma reesei

[0207] Meroterpenoid cyclase A0A2M7NI22 Flexibacter sp.

[0208] CG_4_1O_14_3_um_filter_32_15 Meroterpenoid cyclase A0A4U3L2A6 Ilyomonas limi

[0209]

[0210] Meroterpenoid cyclase A0A2K8Z520 Spirosoma pollinicola Meroterpenoid cyclase MGYP001404307 Unknown

[0211] 206

[0212] Meroterpenoid cyclase WP_276134691.1 Polluticoccus soli Meroterpenoid cyclase MBS1653346.1 Bacteroidetes bacterium Meroterpenoid cyclase A0A1Q3WC77 Spirosoma sp. 48-14 Meroterpenoid cyclase A0A3N5Q208 Ignavibacteriota bacterium Meroterpenoid cyclase MGYP001618046 Unknown

[0213] 589

[0214] Meroterpenoid cyclase MGYP001553196 Unknown

[0215] 604

[0216] Meroterpenoid cyclase A0A5M6D2E1 Adhaeribacter rhizoryzae Meroterpenoid cyclase A1ZCG1 Microscilla marina ATCC 23134 Meroterpenoid cyclase A0A5R9L592 Dyadobacter luticola Meroterpenoid cyclase A0A7G5XG82 Lacibacter sediminis Meroterpenoid cyclase MGYP003332535 Unknown

[0217] 901

[0218] Meroterpenoid cyclase MGYP001605612 Unknown

[0219] 233

[0220] Meroterpenoid cyclase MFI5185720.1 Chitinophagales bacterium Meroterpenoid cyclase MCD6064911.1 Flavipsychrobacter sp.

[0221] Meroterpenoid cyclase MES1221103.1 Bacteroidota bacterium Meroterpenoid cyclase WP_200714303.1 Mutant of SEQ ID NO: 48 with A126S _A126S

[0222] Meroterpenoid cyclase WP_200714303.1 Mutant of SEQ ID NO: 48 with G123M _G123M

[0223] Meroterpenoid cyclase WP_200714303.1 Mutant of SEQ ID NO: 48 with P165G _P165G

[0224] Meroterpenoid cyclase WP_317769678.1 Mutant of SEQ ID NO: 71 with A123M

[0225] _A123M

[0226]

[0227] Meroterpenoid cyclase WP_317769678.1 Mutant of SEQ ID NO: 71 with A126S _A126S

[0228] Meroterpenoid cyclase WP_317769678.1 Mutant of SEQ ID NO: 71 with P165G _P165G

[0229] Meroterpenoid cyclase WP_318017018.1 Mutant of SEQ ID NO: 73 with A126S _A126S

[0230] Meroterpenoid cyclase WP_318017018.1 Mutant of SEQ ID NO: 73 with G123M _G123M

[0231] Meroterpenoid cyclase WP_318017018.1 Mutant of SEQ ID NO: 73 with P165G _P165G

[0232] Meroterpenoid cyclase WSW93534.1 Mutant of SEQ ID NO: 74 with A126S _A126S

[0233] Meroterpenoid cyclase WSW93534.1 Mutant of SEQ ID NO: 74 with G123M _G123M

[0234] Meroterpenoid cyclase WSW93534.1 Mutant of SEQ ID NO: 74 with P165G _P165G

[0235] Meroterpenoid cyclase WSX12386.1 Mutant of SEQ ID NO: 75 with A126S _A126S

[0236] Meroterpenoid cyclase WSX12386.1 Mutant of SEQ ID NO: 75 with G123M _G123M

[0237] Meroterpenoid cyclase WSX12386.1 Mutant of SEQ ID NO: 75 with P165G _P165G

[0238] Meroterpenoid cyclase WSY54005.1 Mutant of SEQ ID NO: 76 with A126S _A126S

[0239] Meroterpenoid cyclase WSY54005.1 Mutant of SEQ ID NO: 76 with G123M _G123M

[0240] Meroterpenoid cyclase WSY54005.1 Mutant of SEQ ID NO: 76 with P165G _P165G

[0241] Meroterpenoid cyclase WTE43000.1 Mutant of SEQ ID NO: 77 with A126S _A126S

[0242] Meroterpenoid cyclase WTE43000.1 Mutant of SEQ ID NO: 77 with G123M

[0243] _G123M

[0244]

[0245] Meroterpenoid cyclase WTE43000.1 Mutant of SEQ ID NO: 77 with P165G _P165G

[0246] Meroterpenoid cyclase WTK72062.1 Mutant of SEQ ID NO: 78 with A123M _A123M

[0247] Meroterpenoid cyclase WTK72062.1 Mutant of SEQ ID NO: 78 with A126S _A126S

[0248] Meroterpenoid cyclase WTK72062.1 Mutant of SEQ ID NO: 78 with P165G _P165G

[0249] Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in squalene cyclases Motif - Conserved motif in bacterial membrane- integrated meroterpenoid cyclases

[0250] Motif - Conserved motif in bacterial membrane- integrated meroterpenoid cyclases

[0251] Motif - Conserved motif in bacterial membrane- integrated meroterpenoid cyclases

[0252] Motif - Conserved motif in bacterial membrane- integrated meroterpenoid cyclases

[0253] Motif - Conserved motif in bacterial membrane- integrated meroterpenoid cyclases

[0254] Motif - Conserved motif in fungal membrane-integrated meroterpenoid cyclases

[0255] Motif - Conserved motif in fungal membrane-integrated meroterpenoid cyclases

[0256] Motif - Conserved motif in bacterial soluble meroterpenoid cyclases

[0257] Motif - Conserved motif in bacterial soluble meroterpenoid cyclases

[0258]

[0259] Motif - Conserved motif in bacterial soluble meroterpenoid cyclases

[0260] RBS - Ribosome binding site Geranylgeraniol CrtE Pantoea agglomerans diphosphate synthase

[0261] Phosphatase PgpB Escherichia coli

[0262] Sclareol synthase SaSCS Synthetic, N-terminal deleted variant

[0263]

[0264] Abbreviations

[0265] ADH alcohol dehydrogenase

[0266] BVMO Baeyer-Villiger Monooxygenase

[0267] bp base pair

[0268] kb kilo base

[0269] DNA deoxyribonucleic acid

[0270] cDNA complementary DNA

[0271] DMAPP dimethylallyl diphosphate

[0272] FMO Flavin Monooxygenase

[0273] FPP farnesyl diphosphate

[0274] GPP geranyldiphosphate

[0275] GGPP geranylgeranyl diphosphate

[0276] GGPS geranylgeranyl diphosphate synthase GC gas chromatograph

[0277] IPP isopentenyl diphosphate

[0278] iMS mass spectrometer / mass spectrometry MVA mevalonic acid

[0279] PP diphosphate, pyrophosphate

[0280] PCR polymerase chain reaction

[0281] RNA ribonucleic acid

[0282] SHC squalene cyclase

[0283] MeroTPS meroterpenoid cyclase

[0284] mRNA messenger ribonucleic acid

[0285] miRNA micro RNA

[0286] siRNA small interfering RNA

[0287] rRNA ribosomal RNA

[0288] tRNA transfer RNA

[0289] TPP terpenyl diphosphate Definitions

[0290] General terms

[0291] For the descriptions herein and the appended claims, the use of “or” means “and / or” unless stated otherwise. Similarly, “comprise”, “comprises”, “comprising”, “include”, “includes”, and “including” are interchangeable and not intended to be limiting.

[0292] It is to be further understood that where descriptions of various embodiments use the term "comprising," those skilled in the art would understand that in some specific instances, an embodiment can be alternatively described using language "consisting essentially of or "consisting of.

[0293] The term “about” indicates a potential variation of ± 25% ofthe stated value, in particular ± 15%, ± 10 %, more particularly ± 5%, ± 2% or ± 1 %.

[0294] The term "substantially" describes a range of values of from about 80 to 100%, such as, for example, 85-99.9%, in particular 90 to 99.9%, more particularly 95 to 99.9%, or 98 to 99.9% and especially 99 to 99.9%.

[0295] “Predominantly” refers to a proportion in the range of above 50%, as for example in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%, particularly in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%; more in particular, in the range of 51 to 99,9%, particularly in the range of 60 to 99,9%, particularly in the range of 70 to 99,9%, particularly in the range of 75 to 99,9%, particularly in the range of 80 to 99,9%, particularly in the range of 85 to 99,9%, particularly in the range of 90 to 99,9%, particularly in the range of 95 to 99,9%, particularly in the range of 96 to 99,9%, particularly in the range of 97 to 99,9%, particularly in the range of 98 to 99,9%, particularly in the range of 99 to 99,9%, particularly in the range of 99,5 to 99,9%.

[0296] A “main product” in the context ofthe present invention designates a single compound or a group of at least 2 compounds, like 2, 3, 4, 5 or more, particularly 2 or 3 compounds, which single compound or group of compounds is “predominantly” prepared by a reaction as described herein, and is contained in said reaction in a predominant proportion (or “relative ratio”) based on the total amount of the product compounds formed by said reaction. Said proportion may be a molar proportion, a weight proportion or, preferably based on chromatographic analytics, an area proportion calculated from the corresponding chromatogram ofthe reaction products.

[0297] In the context of the invention, the term “relative ratio” (expressed in %) refers to the proportion of a given compound based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) in a reaction. This proportion corresponds to the area ratio derived from the corresponding GC-FID chromatogram. The calculation considers only those product compounds (11-1), (II-2), (I I-3) and / or (II-4) that were identified and quantified based on GC-FID analysis. Compounds not identified and / or below the limit of quantification (designated as “not quantified” or “n.q” in the Examples) were excluded from the determination of the relative ratio (%).

[0298] A “side product” in the context of the present invention designates a single compound or a group of at least 2 compounds, like 2, 3, 4, 5 or more, particularly 2 or 3 compounds, which single compound or group of compounds is not “predominantly” prepared by a reaction as described herein.

[0299] In the context of the invention, the term “relative enzyme activity” (expressed in %) refers to the activity of a given enzyme compared to the best-performing enzyme (defined as 100%) under same conditions. The relative enzyme activity (%) is calculated as the proportion of the total quantified product compounds (II-1), (II-2), (II-3) and / or (II-4) obtained with the given enzyme relative to the total quantified product compounds (II-1), (II-2), (II-3) and / or (II-4) obtained with the best-performing enzyme. Compounds below the limit of quantification (designated as “not quantified” or “n.q” in the Examples) were excluded from this calculation. This proportion corresponds to the area ratio derived from the corresponding GC-FID chromatogram.

[0300] The term "stereoisomers" includes conformational isomers and in particular configuration isomers.

[0301] Included in general are, according to the invention, all “stereoisomeric forms” of the compounds described herein, such as “constitutional isomers” and “stereoisomers”.

[0302] “Stereoisomeric forms” encompass in particular, “stereoisomers” and mixtures thereof, e.g. configuration isomers (optical isomers), such as enantiomers, or geometric isomers (diastereomers), such as E- and Z-isomers, and combinations thereof. If one or more asymmetric centers are present in one molecule, the invention encompasses all combinations of different conformations of these asymmetry centers, e.g. enantiomeric pairs.

[0303] “Stereoselectivity” describes the ability to produce a particular stereoisomer of a compound in a stereoisomerically pure form or to specifically convert a particular stereoisomer in an enzyme catalyzed method as described herein out of a plurality of stereoisomers. More specifically, this means that a product of the invention is enriched with respect to a specific stereoisomer, or an educt may be depleted with respect to a particular stereoisomer. This may be quantified via the purity %ee-parameter calculated according to the formula:

[0304] %ee = [XA-XB] / [ XA+XB]*100,

[0305] wherein XA and XB represent the molar ratio (Molenbruch) of the stereoisomers A and B.

[0306] The term “selectively converting” or “increasing the selectivity” in general means that a particular stereoisomeric form, as for example the E-form, of an unsaturated hydrocarbon, is converted in a higher proportion or amount (compared on a molar basis) than the corresponding other stereoisomeric form, as for example Z-form, either during the entire course of said reaction (i.e. between initiation and termination of the reaction), at a certain point of time of said reaction, or during an “interval” of said reaction. In particular, said selectivity may be observed during an “interval” corresponding 1 to 99%, 2 to 95%, 3 to 90%, 5 to 85%, 10 to 80%, 15 to 75%, 20 to 70%, 25 to 65%, 30 to 60%, or 40 to 50% conversion of the initial amount of the substrate. Said higher proportion or amount may, for example, be expressed in terms of:

[0307] . a higher maximum yield of an isomer observed during the entire course of the reaction or said interval thereof;

[0308] . a higher relative amount of an isomer at a defined % degree of conversion value of the substrate; and / or

[0309] . an identical relative amount of an isomer at a higher % degree of conversion value;

[0310] each of which preferably being observed relative to a reference method,

[0311] said reference method being performed under otherwise identical conditions with known chemical or biochemical means.

[0312] Generally, also comprised in accordance with the invention are all “isomeric forms” of the compounds described herein, such as constitutional isomers and in particular stereoisomers and mixtures of these, such as, for example, optical isomers or geometric isomers, such as E- and Z-isomers, and combinations of these. If several centers of asymmetry are present in a molecule, then the invention comprises all combinations of different conformations of these centers of asymmetry, such as, for example, pairs of enantiomers, or any mixtures of stereoisomeric forms.

[0313] “Yield" and I or the "conversion rate" of a reaction according to the invention is determined over a defined period of, for example, 4, 6, 8, 10, 12, 16, 20, 24, 36 or 48 hours, in which the reaction takes place. In particular, the reaction is carried out under precisely defined conditions, for example at “standard conditions” as herein defined.

[0314] The different yield parameters (" Yield" or Yp / s; " Specific Productivity Yield"; or Space-Time- Yield (STY)) are well known in the art and are determined as described in the literature.

[0315] " Yield" and " Yp / s" (each expressed in mass of product produced / mass of material consumed) are herein used as synonyms.

[0316] The specific productivity-yield describes the amount of a product that is produced per h and L fermentation broth per g of biomass. The amount of wet cell weight stated as WCW describes the quantity of biologically active microorganism in a biochemical reaction. The value is given as g product per g WCW per h (i.e. g / gWCW-1h-1). Alternatively, the quantity of biomass can also be expressed as the amount of dry cell weight stated as DCW. Furthermore, the biomass concentration can be more easily determined by measuring the optical density at 600 nm (OD600) and by using an experimentally determined correlation factor for estimating the corresponding wet cell or dry cell weight, respectively.

[0317] The terms "purified", "substantially purified", and "isolated" as used herein refer to the state of being free of other, dissimilar compounds with which a compound of the invention is normally associated in its natural state, so that the "purified", "substantially purified", and "isolated" subject comprises at least 0.5%, 1%, 5%, 10%, or 20%, or at least 50% or 75% of the mass, by weight, of a given sample. In one embodiment, these terms refer to the compound of the invention comprising at least 95, 96, 97, 98, 99 or 100%, of the mass, by weight, of a given sample. As used herein, the terms "purified," "substantially purified," and "isolated" when referring to a nucleic acid or protein, or nucleic acids or proteins, also refers to a state of purification or concentration different than that which occurs naturally, for example in an prokaryotic or eukaryotic environment, like, for example in a bacterial or fungal cell, or in the mammalian organism, especially human body. Any degree of purification or concentration greater than that which occurs naturally, including (a) the purification from other associated structures or compounds or (b) the association with structures or compounds to which it is not normally associated in said prokaryotic or eukaryotic environment, are within the meaning of "isolated”. The nucleic acid or protein or classes of nucleic acids or proteins, described herein, may be isolated, or otherwise associated with structures or compounds to which they are not normally associated in nature, according to a variety of methods and processes known to those of skill in the art.

[0318] Biochemical and biological terms

[0319] The term “enzymatically catalyzed” or “biocatalytic” method means that said method is performed under the catalytic action of an enzyme, including enzyme mutants. Thus, the method can either be performed in the presence of said enzyme in isolated (purified, enriched) or crude form or in the presence of a cellular system, in particular, natural or recombinant microbial cells containing said enzyme in active form, and having the ability to catalyze the conversion reaction as disclosed herein.

[0320] The term "domain" refers to a set of amino acids or a partial sequence of amino acids residues conserved at specific positions along an alignment of sequences of evolutionarily related proteins. While amino acids at other positions can vary between protein homologues, amino acids that are highly conserved at specific positions of such domain indicate amino acids that are likely essential in the structure, stability or function of a protein. Identified by their high degree of conservation in aligned sequences of a family of protein homologues, they can be used as identifiers to determine if any polypeptide in question belongs to a previously identified polypeptide family.

[0321] The term "motif" or consensus sequence" or "signature" refers to a short-conserved region in the sequence of evolutionarily related proteins. Motifs are frequently highly conserved parts of domains but may also include only part of the domain. Signatures are predictive models which describe protein families, domains or sites. The sequences of motifs can be described using the standard IUPAC one-letter codes for the amino acids. Ambiguities are indicated by listing the acceptable amino acids for a given position between brackets. For example, [LWI] stands for L (Leucine), W (Tryptophan) or I (Isoleucine). X represent positions where independently of each other any natural amino acid residue is present.

[0322] A “protein family” is defined as a group of proteins that share a common evolutionary origin reflected by their related functions, similarities in sequence, or similar primary, secondary or tertiary structure. Proteins within protein families are usually homologous and have similar structure of conserved functional domains and motifs.

[0323] Specialist databases exist for the identification of protein domains, for example, SMART (http: / / smart.embl-heidelberg.de / smart / set_mode.cgi? GENOMIC=1) (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic et al. (2020) Nucleic Acids Res 49, D458-D460), InterPro (Paysan-Lafosse et al, Nucleic Acids Research, Nov 2022; Mulder et al., (2003) Nucl. Acids. Res. 31, 315-318), or Pfam (Bateman et al., Nucleic Acids Research 30(1): 276-280 (2002)).

[0324] Useful tools to search or predict protein domains or protein family signatures in protein sequence are for example the NCBI conserved domain search tool (https: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi) or the InterProScan tool (http: / / www.ebi.ac.uk / interpro / search / sequence / ). Domains or motifs may also be identified using routine techniques, such as by sequence alignment.

[0325] The term " Pfam" refers to a large collection of protein domains and protein families maintained by the Pfam Consortium and available at several sponsored world wide web sites, such as the InterPro consortium web site https: / / www.ebi.ac.uk / interpro / (European Molecular Biology Laboratory- Europe an Bioinformatics Institute (EMBL_EBI). The latest release of Pfam is Pfam 35.0 (November 2021), based on the UniProt Reference Proteomes (El-Gebali S. et al, 2019, Nucleic Acids Res. 47, Database issue D427-D432). Pfam domains and families are identified using multiple sequence alignments and hidden Markov models (HMMs). Pfam-A family or domain assignments, are high quality assignments generated by a curated seed alignment using representative members of a protein family and profile hidden Markov models based on the seed alignment (Unless otherwise specified, matches of a queried protein to a Pfam domain or family are Pfam-A matches). All identified sequences belonging to the family are then used to automatically generate a full alignment for the family (Sonnhammer (1998) Nucleic Acids Research 26, 320-322; Bateman (2000) Nucleic Acids Research 26, 263-266; Bateman (2004) Nucleic Acids Research 32, Database Issue, D138-D141; Finn (2006) Nucleic Acids Research Database Issue 34, D247-251; Finn (2010) Nucleic Acids Research Database Issue 38, D211-222). By accessing the Pfam database, for example, using any of the above- reference websites, protein sequences can be queried against the HMMs using HMMER homology search software (e.g., HMMER2, HMMER3, or a higher version, hmmer.janelia.org / ). Significant matches that identify a queried protein as being in a pfam family (or as having a particular Pfam domain) are those in which the bit score is greater than or equal to the gathering threshold for the Pfam domain. Expectation values (E-values) can also be used as a criterion for inclusion of a queried protein in a Pfam or for determining whether a queried protein has a particular Pfam domain, where low e-values, much less than 1.0, for example less than 0.1, or less.

[0326] The term “InterPro” refers to a resource that provides functional analysis of protein sequences by classifying them into families and predicting the presence of domains and important sites (Paysan-Lafosse T, Blum M, Chuguransky S, Grego T, Pinto BL, Salazar GA, Bileschi ML, Bork P, Bridge A, Colwell L, Gough J, Haft DH, Letunic I, Marchler-Bauer A, Mi H, Natale DA, Orengo CA, Pandurangan AP, Rivoire C, Sigrist CJA, Sillitoe I, Thanki N, Thomas PD, Tosatto SCE, Wu CH, Bateman A. InterPro in 2022. Nucleic Acids Research, Nov 2022, (doi: 10.1093 / nar / gkac993)). To classify proteins in this way, InterPro uses predictive models, known as signatures, provided by several collaborating databases (referred to as member databases) that collectively make up the InterPro consortium. A key value of InterPro is that it combines protein signatures from these member databases into a single searchable resource, capitalising on their individual strengths to produce a powerful integrated database and diagnostic tool. Additionally, InterPro adds value to the entries by providing detailed functional annotation as well as adding relevant GO terms that enable automatic annotation of millions of GO terms across the protein sequence databases. InterPro integrates signatures from the following 13 member databases: CATH, CDD, HAMAP, MobiDB Lite, Panther, Pfam, PIRSF, PRINTS, Prosite, SFLD, SMART, SUPERFAMILY and NCBIfam. The member databases use a variety of different methods to classify proteins. Each of the databases has a particular focus (e.g. protein domains defined from structure, or full-length protein families with shared function). InterPro integrates the signatures from the member databases into InterPro entries and identifies where different member database entries are the same entity. InterPro website (https: / / www.ebi.ac.uk / interpro / ) can be used to obtain information about individual protein families, domains, important sites, perform a sequence search or browse through InterPro annotations. InterPro is updated approximately every 8 weeks. InterPro website also provides InterProScan tool (https: / / www.ebi.ac.uk / interpro / about / interproscan / ), a software package that allows sequences to be scanned against InterPro's member database signatures. Users who have novel nucleotide or protein sequences that they wish to functionally characterise can use InterProScan to run the scanning algorithms against the InterPro database in an integrated way. In InterPro, a protein family is defined as a group of proteins that share a common evolutionary origin reflected by their related functions, similarities in sequence, or similar primary, secondary or tertiary structure. A match to an InterPro, or InterPro ID, entry of this type indicates membership of a protein family.

[0327] The “E-value” (expectation value) is the number of hits that would be expected to have a score equal to or better than this value, by chance alone. This means that a good E-value which gives a confident prediction is much less than 1. E-values around 1 is what is expected by chance. Thus, the lower the E-value, the more specific the search for domains will be. Only positive numbers are allowed.

[0328] A “precursor” compound or molecule of a target compound or molecule as described herein is converted to said target compound, preferably through the enzymatic action of a suitable polypeptide performing at least one structural or functional change on said precursor molecule. For example, a “non-cyclic precursor” (like a “non-cyclic terpenyl precursor”) may be converted to the cyclic target molecule (like a cyclic terpene compound) through the action of a cyclase or synthase enzyme, irrespective of the particular enzymatic mechanism of such enzyme, in one or more steps.

[0329] The enzyme nomenclature or enzyme classification (EC) established by the International Union of Biochemistry and Molecular Biology (IUBMB) is a system of naming and categorizing enzymes based on their catalytic activity and biochemical properties. The enzyme nomenclature is widely used in biochemistry to classify and categorize based on their function. The E. C. classification assigns each enzyme a number reflecting the reaction or the type of reaction catalyzed by this enzyme.

[0330] The enzyme classification can be explored using the ‘ExplorEnz’ database (https: / / www. enzymedatabase. org / ) or International Union of Biochemistry and Molecular Biology (IUBMB) web site (https: / / iubmb.qmul.ac.uk). Information can be found about the classification and nomenclature of enzymes, their functions and properties. The database can be searched to find information for a specific enzyme family or enzyme.

[0331] The term “biological function,” “function”, “biological activity” or “activity” of a terpenyl-disphosphate synthase refers to the ability of a terpenyl-diphosphate synthase as described herein to catalyze the formation of at least one terpenyl diphosphate compound from the corresponding precursor terpene.

[0332] The term “biological function,” “function”, “biological activity” or “activity” of a terpenyl-diphosphate phosphatase refers to the ability of the terpenyl-diphosphate phosphatase as described herein to catalyze the removal of a diphosphate group from said terpenyl disphophate compound (precursor) to form the corresponding terpene alcohol.

[0333] The term "homologous" or “endogenous” when used to indicate the relation between a given (recombinant) polynucleotide (such as DNA or RNA) or polypeptide and a given host organism or host cell such as the cell as disclosed herein, is understood to mean that in nature the polynucleotide or polypeptide molecule is produced by a host cell or organism of the same species, such as of the same variety or strain.

[0334] The term "heterologous" when used with respect to a polynucleotide (such as DNA or RNA) or a polypeptide refers to a polynucleotide or polypeptide that does not occur naturally as part of the host cell such as the cell as disclosed herein. In other words, heterologous polynucleotides or polypeptides are not endogenous to the cell into which they are introduced but have been obtained from another cell or synthetically or recombinantly produced.

[0335] As used herein, the term “host cell”, “recombinant cell” or “transformed cell” refers to a cell (or organism) altered to harbor at least one nucleic acid molecule, for instance, a recombinant gene encoding a desired protein or nucleic acid sequence which upon transcription yields at least one functional polypeptide of the present invention. The host cell is particularly a bacterial cell, a fungal cell or a plant cell or plants. The host cell may contain a recombinant gene or several genes, as for example organized as an operon, which has been integrated into the nuclear organelle genomes of the host cell. Alternatively, the host may contain the recombinant gene extra-chromosomally. Methods of introducing recombinant nucleic acid sequences into such host cells are well known in the art and constitute routine laboratory methodologies which do not need to be further described herein.

[0336] The term “organism” refers to any non-human multicellular or unicellular organism such as a plant, or a microorganism. Particularly, a micro-organism is a bacterium, a yeast, an algae or a fungus.

[0337] The term “plant” is used interchangeably to include plant cells including plant protoplasts, plant tissues, plant cell tissue cultures giving rise to regenerated plants, or parts of plants, or plant organs such as roots, stems, leaves, flowers, pollen, ovules, embryos, fruits and the like. Any plant can be used to carry out the methods of an embodiment herein.

[0338] The “mevalonate pathway” also known as the “isoprenoid pathway” or “HMG-CoA reductase pathway” is an essential metabolic pathway present in eukaryotes, archaea, and some bacteria. The mevalonate pathway begins with acetyl-CoA and produces two five-carbon building blocks called isopentenyl pyrophosphate (IPP) and dimethyl allyl pyrophosphate (DMAPP). Key enzymes are acetoacetyl-CoA thiolase (atoB), HMG-CoA synthase (mvaS), HMG-CoA reductase (mvaA), mevalonate kinase (MvaK1), phosphomevalonate kinase (MvaK2), a mevalonate diphosphate decarboxylase (MvaD), and an isopentenyl diphosphate isomerase (idi). Combining the mevalonate pathway with enzyme (prenyltransferase) activity to generate the terpene precursors GPP, FPP or GGPP, like in particular FPP synthase (ERG20), allows the recombinant cellular production of terpenes.

[0339] The term “prenyltransferase”, “prenyltransferase enzyme” or “polypeptide having prenyltransferase activity” represents a group of enzymes having the ability to condense successively five-carbon units such as isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) to form non-cyclic (linear) terpenyl-diphosphate compounds such as geranyl-diphosphate (GPP), farnesyl-diphosphate (FPP) or geranylgeranyl-diphosphate (GGPP) containing 10, 15 and 20 carbons, respectively. Some prenyltransferases can add 5-carbon units to linear terpenyl-diphosphate compounds thereby extending the carbon chain length. An example of prenyl transferase are farnesyl diphosphate (FPP) synthases (FPP synthases; EC 2.5.1.10) having the ability of producing FPP from IPP and DMAPP. Another example of prenyl transferase are geranyl-diphosphate synthases (GGPP synthases; EC 2.5.1.29) having the ability of producing GGPP from IPP and DMAPP or by adding 5 carbons to FPP.

[0340] The term “beta-farnesene synthase”, “beta-farnesene synthase enzyme” or “polypeptide having beta-farnesene synthase activity” (EC 4.2.3.47) represents a group of enzymes having the ability to cleave the diphosphate moiety and a subsequent rearrangement of the farnesyl carbocation intermediate, resulting in the formation of beta-farnesene. Structurally, beta-farnesene synthase belongs to the family of terpene synthases, characterized by conserved motifs such as the DDXXD and NSE / DTE triads, which coordinate divalent metal ions necessary for enzymatic activity. The term “terpene cyclase”, “terpene cyclase enzyme” or “polypeptide having terpene cyclase activity” represents a group of enzymes that catalyze the cylization of terpene precursors into cyclic terpene compounds. Terpene cyclases are divided into two categories depending on the way the initial carbocation is generated. In class I (or type I) terpene cyclase, the diphosphate group of the linear terpenoid precursor is abstracted to form an allylic carbocation on the terpene moiety. In class II, the initial carbocation is formed by protonation of a double bond or epoxy group in the terpene carbon chain. Thus, class I cyclase necessarily use substrates with a diphosphate group, while class II cyclase (since they do not need a diphosphate group for the generation of the initial carbocation) can use terpenoids as substrates.

[0341] In the context of the invention, the term “squalene cyclase (SHC)”, “SHC enzyme”, or “polypeptide having squalene cyclase activity” relates to a polypeptide having terpene cyclase activity wherein the substrate does not contain a diphosphate functional group. The squalene cyclase enzyme family comprises squalene cyclases and 2,3-oxidosqualene cyclases and enzymes catalyzing mechanistically related cyclization reactions. Squalene cyclases catalyze a protonation-initiated cyclization cascade of a linear terpene to a cyclic compound. The squalene cyclase enzyme family includes for example squalene-hopene cyclases catalyzing the cyclization of squalene to hopene (EC 5.4.99.17) and squalene-hopanol cyclases catalyzing the cyclization of squalene to hopan-22-ol (EC 4.2.1.129). Tetraprenyl-p-curcumene-sporulenol cyclases catalyze similar cyclization of linear terpene substrate (EC 4.2.1.137). It was shown that tetraprenyl-p-curcumene-sporulenol cyclase can also catalyze the cyclization of squalene (Sato, T., et al. (2011). Journal of the American Chemical Society 133(44): 17540-17543), thus tetraprenyl-p-curcumene-sporulenol cyclases are also members of the squalene cyclase family.

[0342] The enzymatic activity of a squalene cyclase (SHC) is determined under “standard conditions” as for example described in the Examples section.

[0343] In the context of the invention, the term “meroterpenoid cyclase (MeroTPS)”, ‘MeroTPS enzyme” or “polypeptide having meroterpenoid cyclase activity” relates to a polypeptide capable of catalyzing the cyclization of meroterpenoids, which are a class of natural products that consist of a terpenoid moiety combined with a non-terpenoid (often aromatic) moiety. Said meroterpenoid cyclases have been described in PCT / EP2024 / 066253 and were categorized into 3 categories, i.e. membrane-integrated meroterpenoid cyclase of bacterial origin, membrane-integrated meroterpenoid cyclase of fungal origin, and soluble meroterpenoid cyclases of bacterial origin.

[0344] The enzymatic activity of a meroterpenoid cyclase is determined under “standard conditions” as for example described in the Examples section.

[0345] Chemical terms:

[0346] Diphosphate” and “pyrophosphate” as used herein are synonyms. “Terpenes” are a large and diverse class of organic compounds, produced by a variety of plants, particularly conifers, and by some insects. Terpenes are hydrocarbons. Although sometimes used interchangeably with "terpenes", “terpenoids” or “isoprenoids” are modified terpenes as they contain additional functional groups, usually oxygen-containing.

[0347] “Terpenoids” (“isoprenoids”) are a large and diverse class of naturally occurring organic chemicals derived from terpenes. Although sometimes used interchangeably with the term “terpenes”, “terpenoids” contain additional functional groups, usually O-containing groups, like for example hydroxyl, carbonyl or carboxyl groups. Most are multicyclic structures with oxygen-containing functional groups. Unless stated otherwise, in the context of the present description the term “terpene” and the term “terpenoid” may be used interchangeably.

[0348] Terpenes (and terpenoids) may be classified by the number of isoprene units in the molecule; a prefix in the name indicates the number of terpene units needed to assemble the molecule. Hemiterpenes consist of a single isoprene unit. Monoterpenes consist of two isoprene units and have the molecular formula C10H16. Sesquiterpenes consist of three isoprene units and have the molecular formula C15H24. Diterpenes are composed of four isoprene units and have the molecular formula C20H32.

[0349] “Terpenyl” designates noncyclic and cyclic chemical hydrocarbyl residues which are derived from the C5building block isoprene and in particular contain one or more such building blocks.

[0350] “Cyclic terpene” or cyclic terpenyl” or “cyclic diterpene” or cyclic diterpenyl” relates to a terpene compound or terpenyl residue which comprises in its structure at lest on, as for example 1, 2, 3, 4 or 5 carbocyclic condensed and / or non-condensed rings, preferably two carbocyclic condensed rings.

[0351] “Bicyclic terpene” or bicyclic terpenyl” or “bicyclic diterpene” or bicyclic diterpenyl” relates to a terpene compound or terpenyl residue which comprises in its structure two carbocyclic rings, preferably two carbocyclic condensed rings.

[0352] “Derivatives of terpenes” or “derivatives of terpenoids” in the context of the present invention in particular refer to such chemical compounds which are obtained from a terpene or terpenoid by chemical and / or enzymatic modification.

[0353] A “hydrocarbyl” residue is a chemical group which essentially is composed of carbon and hydrogen atoms and may be a non-cyclic, linear or branched, saturated or unsaturated moiety, or a cyclic saturated or unsaturated moiety, aromatic or non-aromatic moiety. A hydrocarbyl residue comprises 1 to 30, 1 to 25, 1 to 20, 1 to 15 or 1 to 10 or 1 to 5 carbon atoms in the case of a non-cyclic structure. It comprises 4 to 30, 4 to 25, 4 to 20, 4 to 15, 4 to 10 or in particular 4, 5, 6 or 7 carbon atoms in the case of a cyclic structure. Said hydrocarbyl residues may be non-substituted or may carry at least one, like 1 to 5, preferably 0, 1 or 2 substituents.

[0354] Particular examples of such hydrocarbyl residues are non-cyclic linear or branched alkyl or alkenyl residues; or mono- or polycyclic, in particular mono- or bicyclic, saturated or unsaturated, nonaromatic moieties, as for example found in cyclic (for example bicyclic) or non-cyclic terpene type compound, and labdane-type compounds.

[0355] In the present document, a “Cx-y-alkyl” group is an alkyl group comprising x to y carbon atoms, i.e., for example, a Ci-3-alkyl group is an alkyl group comprising 1 to 3 carbon atoms. The alkyl group can be linear or branched. For example -CH(CH3)-CH2-CH3 is considered as a C4-alkyl group.

[0356] An “alkenyl” residue represents linear or branched, mono- or polyunsaturated hydrocarbon residues. It comprises 2 to 30, 2 to 25, 2 to 20, 2 to 15 or 2 to 10 or 2 to 7, 2 to 6, 2 to 5, or 2 to 4 carbon atoms. I may have up to 10, like 1, 2, 3, 4 or 5 C=C double bonds.

[0357] An "alkylene" represents straight-chain or singly or multiply branched hydrocarbon bridging groups having 1 to 10 carbon atoms, for example Ci-C -alkylene groups selected from -CH2-, -(CH2)2-, -(CH2)3-,-(CH2)4-, -(CH2)2-CH(CH3)-, -CH2-CH(CH3)-CH2-, (CH2)4-, -(CH2)5-, -(CH2)6, -(CH2)7-, -CH(CH3)-CH2-CH2-CH(CH3)- or -CH(CH3)-CH2-CH2-CH2-CH(CH3)-, and in particular C1-C4-alkylene groups selected from -CH2-, -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)2-CH(CH3)-, -CH2-CH(CH3)-CH2-.

[0358] An “alkylidene” group represents a straight chain or branched hydrocarbon substituent linked via a double bond to the body of the molecule. It comprises 1 to 6 carbon atoms. As examples of such “C1-Ce-alkylidenes” there may be mentioned methylidene (=CH2) ethylidene, (=CH-CH2), n- propylidene, n-butylidene, n-pentlyiden, n-hexylidene and the constitutional isomers thereof, as for example iso-propylidene.

[0359] An “alkenylidene” represents the mono-unsaturated analogue of the above mentioned alkylidenes with more than 2 carbon atoms and may be called “Cs-Ce-alkenylidenes”. n- propenylidene, n-butenylidene, n-pentenlyiden, and n-hexenylidene may be mentioned as examples.

[0360] In case identical labels for symbols or groups are present in several formulae, in the present document, the definition of said group or symbol made in the context of one specific formula applies also to other formulae which comprises the same said label.

[0361] The term “independently from each other” in this document means, in the context of substituents, moieties, or groups, that identically designated substituents, moieties, or groups can occur simultaneously with a different meaning in the same molecule.

[0362] Any single dotted line in any formulae represents the bond by which said substituent is bound to the rest of a molecule.

[0363] Any wavy line in any formula of this document represents a carbon-carbon bond which when linked to the carbon-carbon double bond is either in the Z- or in the E-configuration. The “substituent” of the above-mentioned residues contains one hetero atom, like O or N. Preferably the substituents are independently selected from -OH, C=O, or - COOH. Most preferably said substituent is -OH.

[0364] A “mono- or polycyclic hydrocarbyl residue” comprise 1, 2 or 3 condensed (anellated) or noncondensed, optionally substituted, saturated or unsaturated hydrocarbon ring groups (or “carbocyclic” groups). Each cycle may comprise independently of each other 3 to 8, in particular 5 to 7, more particularly 6 ring carbon atoms. As examples of monocyclic residues there may be mentioned "cycloalkyl" groups which are carbocyclic radicals having 3 to 7 ring carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl; and the corresponding “cycloalkenyl” groups. Cycloalkenyl" (or "mono- or polyunsaturated cycloalkyl") represents, in particular, monocyclic, mono- or polyunsaturated carbocyclic groups having 5 to 8, preferably up to 6, carbon ring members, for example monounsaturated cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenylradicals.

[0365] As examples of polycyclic residues there may be mentioned groups wherein 1, 2 or 3 of such cycloalkyl and / or cycloalkenyl are linked together, as for example anellated, in order to form a polycyclic cycloalkyl or cycloalkenyl ring. As non-limiting example the bicyclic decalinyl residue composed of two anellated 6-membered carbon rings may be mentioned.

[0366] The number of substituents in such mono- or polycyclic hydrocarbyl residues may vary from 1 to 10, in particular 1 to 5 substituents. Suitable substituents of such cyclic residues are selected from lower alkyl, lower alkenyl, alkylidene, alkenylidene, or residues containing one hetero atom, like O or N as for example -OH or - COOH. In particular, the substituents are independently selected from -OH, - COOH, methyl and methylidene.

[0367] Unsaturated cyclic groups may contain 1 or more, as for example 1, 2 or 3 C=C bonds and are aromatic, or in particular nonaromatic.

[0368] The above-mentioned mono- or polycyclic saturated or unsaturated groups may also contain at least one, like 1, 2, 3 or 4 ring heteroatoms, such as O, N or S.

[0369] The term "alcohol protecting group" in this document means a group which protects the hydroxyl in any of the formulas in this document and which can be easily removed, (i.e. deprotected), by state-of-the-art methods, resulting to the respective compound with the free hydroxyl group.

[0370] The alcohol protecting group is introduced by a chemical reaction of the compound of the respective formula having OH with a protecting agent.

[0371] The protecting agents leading to the corresponding alcohol protecting groups are known to the person skilled in the art, as well as the chemical process and conditions for this reaction.

[0372] If, for example, the alcohol protecting group forms an ester with the rest of the molecule, the suitable protecting agent is for example an acid, an anhydride, or an acyl halide. If the alcohol protecting group forms an acetal or a ketal with the rest of the molecule, the suitable protecting agent is an aldehyde or a ketone.

[0373] If the alcohol protecting group forms an ether with the rest of the molecule, the suitable protecting agent is an alkyl halide, e.g. MeO(CH2)2OCH2Cl, or an enol ether, e.g. 3,4-dihydro-2 / 7-pyran.

[0374] The preferred alcohol protecting group is an acyl group, particularly a group of the formula

[0375] O

[0376]

[0377] ' R3as defined later on in this document.

[0378] If the present disclosure refers to features, parameters and ranges thereof of different degree of preference (including general, not explicitly preferred features, parameters and ranges thereof) then, unless otherwise stated, any combination of two or more of such features, parameters and ranges thereof, irrespective of their respective degree of preference, is encompassed by the disclosure of the present description. Detailed Description

[0379] The present inventors sought to identify new methods and compounds toward the preparation of perfumery, flavor or aroma compounds. In this context, the present inventors surprisingly found that meroterpenoid cyclases could be used in the cyclisation of linear terpenoid compounds to produce a compound of formula (II-1), (II-2), (II-3) and / or (II-4).

[0380] Accordingly, the present invention provides a novel method for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4) starting from compound of the formula (I). The compounds of the formula (11-1-1), (11-1-1 a), (11-1-2), (ll-1-2a), (II-1-3), (ll-1-3a), (11-1-4), (II-1-4a), (11-1-5), (ll-1-5a), (ll-4-6a), (II-Aa), (ll-1b), (ll-1c) and (ll-1d) are also part of the invention. Said compounds may be used as a perfumery, flavor or aroma ingredient and / or a precursor thereof.

[0381] Method of the invention

[0382] A first aspect of the invention provides a method for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4),

[0383] (II-2)

[0384]

[0385] (H-3)

[0386] (H-4)

[0387]

[0388] and / or

[0389] in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises more than one--OH group,

[0390] a compound having at least one-_O-- group,

[0391] which is formally obtained by an intramolecular etherification or esterification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification or esterification, two--OH groups form, under cleavage of a molecule of water, a cyclic-_O-- group; and / or

[0392] in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises at least two--OH groups and at least one carbonyl group,

[0393] a compound having at least one-_O-- group,

[0394] which is formally obtained by ketalization of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the ketalization a carbonyl group and two--OH groups form, under cleavage of water, a cyclic-_O-- group;

[0395] and / or

[0396] in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises at least one--OH group and at least one carbonyl group,

[0397] a compound having at least one-_O-- group, which is formally obtained by etherification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification an--OH group a carbonyl group having undergone enolization form, under cleavage of water, a cyclic-_O-- group; or a derivative thereof,

[0398] with the proviso that said compound is not

[0399]

[0400] wherein

[0401] A represents either

[0402]

[0403] A' represents H, or

[0404] A' represents OH, in case A represents

[0405]

[0406] R represents

[0407] P l ^°'

[0408]

[0409] wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;

[0410] wherein R2= H or an alcohol protecting group, particularly

[0411]

[0412] R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and

[0413] m = 0 or 1 or 2;

[0414] n = 0 or 1 or 2;

[0415] with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;

[0416] z = 1 or 2; R° independently form each other represents H or CH3or CH2OH;

[0417] wherein

[0418] any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;

[0419] and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond;

[0420] characterized in that the method comprises:

[0421] (a) contacting a compound of the formula (I)

[0422]

[0423] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-1), (II-2), (II-3) and / or (II-4).

[0424] In particular, the invention provides a method for preparing a compound of the formula (11-1), (II-), (II-3) and / or (II-4),

[0425] (II-1)

[0426]

[0427] 2024P00180WO

[0428] (II-2)

[0429] (II-3)

[0430] (II-4)

[0431]

[0432] and / or

[0433] in case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises more than one--OH group,

[0434] compound having at least one-_O-- group,

[0435] which is formally obtained by an intramolecular etherification or esterification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification or esterification, two--OH groups form, under cleavage of a molecule of water, a cyclic-_O-- group;

[0436] with the proviso that said compound is not

[0437]

[0438] or a derivative thereof,

[0439] wherein

[0440] A represents either

[0441]

[0442] A' represents H, or

[0443] R represents

[0444]

[0445] wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;

[0446] wherein R2= H or an alcohol protecting group, particularly

[0447]

[0448] ' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and

[0449] m = 0 or 1 or 2;

[0450] n = 0 or 1 or 2;

[0451] with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;

[0452] z = 1 or 2;

[0453] R° independently form each other represents H or CH3or CH2OH;

[0454] wherein

[0455] any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;

[0456] and wherein any double bond having dotted line ( ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line (

[0457]

[0458] ) represents a carbon-carbon double bond;

[0459] characterized in that the method comprises:

[0460] (a) contacting a compound of the formula (I)

[0461] R°

[0462]

[0463] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-1), (II-2), (II-3) and / or (II-4).

[0464] An embodiment of the method of the invention is characterized in that the cyclic-_O-- group is part of a 4-6 membered ring.

[0465] An embodiment of the method of the invention is characterized in that the cyclic-_O-- group is part of a cyclic ether or a cyclic ester (lactone).

[0466] An embodiment of the method of the invention is wherein the compound having at least one —O— group is selected from the group consisting of the compounds of the formula

[0467]

[0468]

[0469] (H-E), and / or

[0470]

[0471] characterized in that the method comprises:

[0472] (a) contacting a compound of the formula (I)

[0473] wherein

[0474] R represents

[0475]

[0476] wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;

[0477] wherein R2= H or an alcohol protecting group, particularly

[0478]

[0479] '' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and

[0480] m = 0 or 1 or 2;

[0481] n = 0 or 1 or 2;

[0482] with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;

[0483] z = 1 or 2; R° independently form each other represents H or CH3or CH2OH;

[0484] wherein

[0485] any dotted line represents the bond by which the substituent is bound to the rest of the molecule

[0486] any wavy line represents a carbon-carbon bond which is either in the Z- or in the E- configuration, preferably in the E-configuration;

[0487] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the meroterpenoid cyclase enzyme to produce the compound of the formula (II-A), (II-B), (II-C), (ll-D), (ll-E) and / or (ll-F).

[0488] In a further embodiment, the invention provides a method for preparing a compound of the formula

[0489]

[0490] characterized in that the method comprises:

[0491] (a) contacting a compound of the formula (I) R°

[0492]

[0493] wherein

[0494] A represents either

[0495]

[0496] R represents

[0497]

[0498] , or

[0499] wherein R1= OH or a Ci-3-alkyl group;

[0500] wherein R2= H; and

[0501] m = 0 or 1 or 2;

[0502] n = 0 or 1 or 2;

[0503] with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;

[0504] R° independently form each other represents H or CH3or CH2OH;

[0505] wherein

[0506] any dotted line represents the bond by which the substituent is bound to the rest of the molecule

[0507] any wavy line represents a carbon-carbon bond which is either in the Z- or in the E- configuration, preferably in the E-configuration;

[0508] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the meroterpenoid cyclase enzyme to produce the compound of the formula (II-A), (II- B), (II-C), (ll-D), (ll-E) and / or (ll-F).

[0509] It is important to realize that by starting from a specific starting material compound of the formula (I), the process yields one or several product compounds of the formula (11-1), (II-2), (II-3) and / or (II-4).

[0510] The starting material compounds of the formula (I) for the present method for preparing a compound of the formula (11-1), (II-2), (II-3) and / or (II-4) are the compounds selected from the group consisting of the compounds of the formula:

[0511] (l-a),

[0512]

[0513] (l-d),

[0514] (l-h),

[0515]

[0516]

[0517] Particularly, it has been found that the process provides specific product compounds of the formula (11-1), (II-2), (II-3) and / or (II-4) from the specific starting material compounds of the formula (I). Said specific starting material compounds of the formula (I) and said specific product compounds of the formula (II-1), (II-2), (II-3) and (II-4) are listed herein below in Table A and Table B. In said Tables, the “Rx” groups correspond to the groups identified elsewhere in the description.

[0518]

[0519]

[0520] 2024P00180WO

[0521] b. Compounds of the formula (II-1), (II-2), (II-3) and (II-4)

[0522]

[0523]

[0524]

[0525]

[0526] 2024P00180WO

[0527]

[0528]

[0529]

[0530] Terpene cyclases in the method of the invention

[0531] Terpene cyclases are divided into two categories depending on the way the initial carbocation is generated. In class I (or type I) terpene cyclase, the diphosphate group of the linear terpenoid precursor is abstracted to form an allylic carbocation on the terpene moiety. In class II, the initial carbocation is formed by protonation of a double bond or epoxy group in the terpene carbon chain. Thus, class I cyclase necessarily use substrates with a diphosphate group, while class II cyclase (since they do not need a diphosphate group for the generation of the initial carbocation) can use terpenoids as substrates.

[0532] For all terpene cyclases, the generated reactive carbocation species triggers the subsequent cascade reaction including carbocation reactions with double bonds, alkyl-shifts, hydride shifts or carbon-carbon bound formation. The reaction can be terminated by deprotonation of a carbon atom adjacent to the carbocation or by quenching of the carbocation with a hydroxyl group or molecule of water.

[0533] The type II activity in terpene cyclases is associated with aspartate-rich conserved motifs.

[0534] Typical examples of class II terpene cyclases are the class II diterpene cyclases catalyzing the protonation-initiated cyclization of geranylgeranyl-diphosphate into for example, labdadienyl-diphosphate intermediates or other cyclic diphosphate intermediates (Peters, R. J. (2010). Nat. Prod. Rep. 27, 1521— 1530; Zerbe, P. et al (2015). Plant J. 83, 783-793).

[0535] Squalene cyclases (SHCs) constitute a classical example of class II terpene cyclases where the substrate does not contain a diphosphate functional group. The squalene cyclase enzyme family comprise squalene cyclases and 2,3-oxidosqualene cyclases and enzymes catalyzing mechanistically related cyclization reactions. Squalene cyclases catalyze a protonation-initiated cyclization cascade of a linear terpene to a cyclic compound. Thus, squalene cyclases are class II terpene cyclases. The squalene family includes for example squalene-hopene cyclases catalyzing the cyclization of squalene to hopene (EC 5.4.99.17) and squalene-hopanol cyclases catalyzing the cyclization of squalene to hopan-22-ol (EC 4.2.1.129). Tetraprenyl-p-curcumene-sporulenol cyclases catalyze similar class II cyclization of linear terpene substrate (EC 4.2.1.137). It was shown that tetraprenyl-p-curcumene-sporulenol cyclase can also catalyze the cyclization of squalene (Sato, T., et al. (2011). Journal of the American Chemical Society 133(44): 17540-17543), thus tetraprenyl-p-curcumene-sporulenol cyclases are also members of the squalene cyclase family.

[0536] Squalene cyclase polypeptides have typically a length between 600 and 800 amino acids and are membrane-associated proteins. They bind to the surface of cellular membranes but do not contain a transmembrane region. Squalene cyclases are classified in the IPR018333 family of the InterPro protein sequence classification database (https: / / www.ebi.ac.uk / interpro / entry / InterPro / IPR018333 / ) (InterPro release 93.0, 2nd March 2023). The structure of squalene cyclases is organized in two domains comprising several alpha-helices, recognized as the β-domain and γ-domain or the βγ-domain architecture (Christianson DW, Chem. Rev, 2017, 117, 11570-11648). The two domains have characteristic sequence signatures as described in the Pfam database under the Pfam Squalene-hopene cyclase N-terminal domain (PF13249) and Squalene-hopene cyclase C-terminal domain and (PF13243) (Pfam 35.0 released, 19 November 2021). The presence of the IPR018333, PF13249 or PF13243 protein sequences signatures can be predicted using the NCBI conserved domain search tool (https: / / www.ncbi.nlm.nih.gov / Structure / cdd / wrpsb.cgi) or the InterProScan tool (http: / / www.ebi.ac.uk / interpro / search / sequence / ).

[0537] The squalene cyclase polypeptide contains characteristic conserved amino acid motifs located along the sequence and associated with the protein architecture and enzymatic reaction. In particular, the squalene cyclase contains at least one or more amino acid motifs selected from:

[0538] ■ [SP][TP][VIL]WDTx[LWI] (SEQ ID NO: 205),

[0539] . PGG[WF][GYA]F (SEQ ID NO: 206),

[0540] . PDxDD[TAS][TIAS] (SEQ ID NO: 207),

[0541] . [MIL]QxxxG[GA][WF]x[AS][FY] (SEQ ID NO: 208),

[0542] . Qxxx[GH]xWxG[RK]WGxx[YF]xYG (SEQ ID NO: 209),

[0543] . Qxx[DN]G[GS][WF][GS]ExxxS (SEQ ID NO: 210), and

[0544] . [STA]xx[SFN][QC]T[AGT]W[AS][LIV]xx[LQ] (SEQ ID NO: 211).

[0545] The motif sequences are described using the standard IUPAC one-letter codes for the amino acids. Ambiguities are indicated by listing the acceptable amino acids for a given position between brackets. For example, [SP] or [S or P] stands for S (serine), or P (proline). The “x” represents positions where independently of each other any natural amino acid residue is present. The function of the square brackets has been described above.

[0546] Meroterpenoids are hybrid secondary metabolites derived from mixed biosynthetic pathways and are partially derived from a terpenoid co-substrate (Cornforth, J. W. Terpenoid biosynthesis. Chem. Br.

[0547] 1968, 4, 102-106). The non-terpenoid part can originate for example from polyketides, alkaloids, phenols, or amino acids biosynthetic pathway. Large chemical diversity is found among meroterpenoids, in particular in bacteria and in fungi.

[0548] The meroterpenoids biosynthetic pathways follow several modular biosynthetic steps. In the first step, the building blocks are generated from the corresponding biosynthetic pathway (e.g. terpenoids, polyketides). The terpenoid and non-terpenoid parts are assembled by prenyltransferases. The precursors of the terpenoid parts are generally linear terpenoid-diphosphates such as geranyldiphosphate, farnesyl-diphosphate orgeranylgeranyl-diphosphate.

[0549] In the following step, the linear polyene terpenoid part of the hybrid precursor is cyclized to form a monocyclic or polycyclic structure. This cyclization is catalyzed by a specific class of non-canonical class II terpene cyclases named meroterpenoid cyclase first discovered in fungi. The first discovered representative meroterpenoid cyclase is Pyr4 from Aspergillus fumigatus Af293 (Itoh, T., et al. (2010). Nature Chemistry 2(10): 858-864).

[0550] In many meroterpenoids, the linear terpenoid precursor is first activated by a stereoselective epoxidation by a monooxygenase of one of the double bonds. The meroterpenoid cyclases catalyze then the protonation of the epoxide moiety generating a reactive carbocation species and triggering a subsequent cascade reaction similar to other terpene cyclases. Some meroterpenoid cyclases can convert the isoprenic precursors to cyclized products without the involvement of a prior epoxidation step. These meroterpenoid cyclases are able to directly protonate the terminal double bond generating a reactive carbocation and catalyzing a cyclization. For example, MacJ from the fungi Penicillium terrestry was the first identified fungi meroterpenoid cyclase using a type II double-bond protonation initiations reaction (Tang, M.-C., et al. (2017). Organic Letters 19(19): 5376-5379). Another example of meroterpenoid cyclase which initiates polyene cyclization by direct double bond protonation is DmtA1 from bacteria (Streptomyces youssoufiensis OUC68199) (Yao et al, Nat. Commun., 2018, 9, 4091).

[0551] Like other class II terpene cyclases, the carbocation generated by meroterpenoid cyclases triggers a cascade reaction generally starting by the attack of a double bond and generating monocyclic or polycyclic structure with a tertiary carbocation. The reaction is terminated either by deprotonation to form a double bond or by reacting with a water molecule to generate a tertiary alcohol. Typical cyclic structures found in meroterpenoids compounds contain drimane or labdane scafolds.

[0552] The largest group of meroterpenoid cyclases are compact membrane-integrated proteins containing several (generally seven) transmembrane helices. This protein architecture based on transmembrane helices can easily be predicted using for example the TMHMM 2.0 server available at https: / / dtu.biolib.com / DeepTMHMM (Krogh, A., et al. (2001) J Mol Biol 305(3): 567-580.). In addition to the protein architecture, meroterpenoid cyclases differ from other class II cyclases such as the squalene cyclases by their smaller polypeptide size. The bacterial and fungal meroterpenoid cyclase polypetides have a length ranging from 150 to 550 residues. The transmembrane helices are located over a portion of the polypetide covering 180 to 300 amino acid and carry the catalytic domains.

[0553] Recently meroterpenoid cyclases having a protein architecture different from the membrane-integrated meroterpenoid cyclases were described. For example, MstE from the bacteria Scytonema sp. PCC 1002 is a soluble cyclase having a structure similar to canonical cyclases such as diterpene synthases and squalene cyclases, but nevertheless different, since it is a monodomain protein with only an a-domain (Moosmann, P., etal. (2020). Nat Chem 12(10): 968-972). Soluble bacterial meroterpenoid cyclase polypetides have lengths ranging from 150 to 550.

[0554] Meroterpenoid cyclase polypeptides contain characteristic conserved amino acid motifs located along the sequence and associated with the protein architecture or enzymatic reaction as follows: Membrane-integrated meroterpenoid cyclase of bacterial origin containing at least one or more amino acid motifs selected from:

[0555] . [W]xxx[D]xx[ILVMN] (SEQ ID NO: 212),

[0556] . PxxAxxxNxxWE (SEQ ID NO: 213),

[0557] . MxxxFxxMLxxR (SEQ ID NO: 214),

[0558] . RxxxxGQS (SEQ ID NO: 215), and

[0559] . NxxMS (SEQ ID NO: 216).

[0560] Membrane-integrated meroterpenoid cyclase of fungal origin containing a least one or more amino acid motifs selected from:

[0561] . [WY]Exx[YFW] (SEQ ID NO: 217), and

[0562] . [DNE]xSYxxP (SEQ ID NO: 218).

[0563] Soluble meroterpenoid cyclases of bacterial origin containing a least one or more amino acid motifs selected from:

[0564] . GxWxxxW[WG]xxxxY (SEQ ID NO: 219),

[0565] . WxxxHxxV[TSA] (SEQ ID NO: 220), and

[0566] . GxWxD[FY] (SEQ ID NO: 221).

[0567] The motif sequences are described using the standard IUPAC one-letter codes for the amino acids. Residues x represent independently of each other any natural amino acid residue, and wherein optionally in each of the above motifs, 1, 2, 3 or 4 amino acid residues different from the x residues may be modified, for example by amino acid substitution, in particular by conservative substitutions, provided that the enzyme retains, at least to analytically detectable extent, its enzyme activity. The function of the square brackets has been described above.

[0568] For the avoidance of doubt, SHCs and meroterpenoid cyclases are distinct classes of enzymes which can be distinguished by physical characteristics. Furthermore, meroterpenoid cyclases can be classified as (i) bacterial membrane-integrated meroterpenoid cyclases; (ii) fungal membrane-integrated meroterpenoid cyclases; (iii) bacterial soluble meroterpenoid cyclases.

[0569] Table C below outlines the differences between SHCs and the different types of meroterpenoid cyclases.

[0570] Feature SHC Bacterial membrane- Fungal membrane- Bacterial soluble integrated meroTPS integrated meroTPS MeroTPS Protein size (# 600-800 150-550 (180-300 150-550 (180-300 150 to 550 amino acids) for region for region containing

[0571] containing the the helices)

[0572] helices)

[0573] Transmembrane No Yes Yes No

[0574] region

[0575] Membrane- membrane- membrane- Soluble associated integrated integrated

[0576] Protein structure βγ-domain transmembrane transmembrane a-domain architecture (2 helices helices architecture domains) (monodomain

[0577] protein)

[0578]

[0579] Table C: enzyme characteristics Hence the skilled person can, from the information provided herein, readily identify whether an enzyme is a SHC enzyme or a class of meroterpenoid cyclase enzyme.

[0580] Several meroterpenoid cyclases catalyze reactions of cyclisation of the terpenoid part of the meroterpenoid hybrid precursor to labdane cyclic structures. However, for the first time, the present inventors surprisingly found that meroterpenoid cyclases could be used for the cyclization of a linear terpenoid (such as a compound of formula (I)) to a compound of the formula (11-1), (II-2), (II-3) and / or (II-4).

[0581] Accordingly, an embodiment of the method of the invention is characterized in that the terpene cyclase is a meroterpenoid cyclase enzyme.

[0582] A further embodiment of the method of the invention is characterized in that the meroterpenoid cyclase enzyme is a bacterial membrane-integrated meroterpenoid cyclase enzyme, preferably comprising at least one or more amino acid motifs selected from:

[0583] [W]xxx[D]xx[ILVMN] (SEQ ID NO: 212),

[0584] PxxAxxxNxxWE (SEQ ID NO: 213),

[0585] MxxxFxxMLxxR (SEQ ID NO: 214),

[0586] RxxxxGQS (SEQ ID NO: 215), and

[0587] NxxMS (SEQ ID NO: 216);

[0588] wherein residues x represent independently of each other any natural amino acid residue.

[0589] Accordingly, in a further embodiment of the method of the invention, the meroterpenoid cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one embodiment, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, the meroterpenoid cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In one embodiment, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180.

[0590] In yet a further embodiment of the invention, the meroterpenoid cyclase enzyme is a mutant meroterpenoid cyclase enzyme, preferably a mutant bacterial membrane-integrated meroterpenoid cyclase enzyme, having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180; wherein the mutant meroterpenoid cyclase enzyme has an amino acid substitution at amino acid position 9 relative to the sequence provided in SEQ ID NO: 29.

[0591] With reference to the above embodiments, the substitution at amino acid position 9 relative to the sequence provided in SEQ ID NO: 29 may be a substitution to cysteine, methionine or threonine. In an alternative or yet a further embodiment of the invention, the meroterpenoid cyclase enzyme is a mutant meroterpenoid cyclase enzyme, preferably a mutant bacterial membrane-integrated meroterpenoid cyclase enzyme, having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180; wherein the mutant meroterpenoid cyclase enzyme has an amino acid substitution at amino acid position 123, 126 and / or 165 relative to the sequence provided in SEQ ID NO: 74.

[0592] With reference to the above embodiments, the substitution at amino acid position 123, 126 and / or 165 relative to the sequence provided in SEQ ID NO: 74 may be a substitution to methionine, serine or glycine.

[0593] Specific embodiments of the method of the first aspect of the invention

[0594] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-a) and the compound of the formula (II-2) is a compound of the formula (II-2-17)

[0595] (l-a),

[0596] (II-2-17);

[0597]

[0598] preferably, a compound of the formula (11-2-17a)

[0599] (11-2-17a).

[0600]

[0601] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 38. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 38.

[0602] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-b) and the compound of the formula (11-1), (II-2), (II-3) and / or (II-4) is a compound of the formula (II-1-3), (II-2-7), (II-2-8), (II-2-9), (II-3-2) and / or (II-4-3)

[0603] (II-1-3),

[0604] (II-2-7),

[0605] (II-2-8),

[0606] (II-2-9),

[0607] (II-3-2),

[0608] (II-4-3);

[0609]

[0610] preferably, a compound of the formula (11-1 -3a), (I l-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (I l-4-3a)

[0611] (ll-1-3a),

[0612] (ll-2-7a),

[0613] (ll-2-8a),

[0614] (II-2-9),

[0615] (II-3-2a),

[0616] (II-4-3a).

[0617]

[0618] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-8, 11-17, 19-22, 25, 28, 29, 32, 35, 36, 38, 39, 41-46, 49-51, 54-62, 64, 65, 67-69, 102-104, 107, 110 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-8, 11-17, 19-22, 25, 28, 29, 32, 35, 36, 38, 39, 41-46, 49-51, 54-62, 64, 65, 67-69, 102-104, 107, 110 and 137-139.

[0619] In a specific embodiment, the compound of the formula (ll-3-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-2a) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (I I-4) (namely, (11-1 -3a), (I l-2-7a), (II-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 8, 14 to 17, 19, 22, 25, 28, 29, 32, 35, 36, 38, 39, 41 to 45, 49 to 51, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 137, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 8, 14 to 17, 19, 22, 25, 28, 29, 32, 35, 36, 38, 39, 41 to 45, 49 to 51, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 137, 138 and 139.

[0620] In another specific embodiment, the compound of the formula (II-2-9) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-2-9) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (II-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 7 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 7 and 46.

[0621] In another specific embodiment, the compound of the formula (ll-2-8a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-2-8a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 11 to 13. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 11 to 13.

[0622] In another specific embodiment, the compound of the formula (11-1 -3a), optionally together with the compound of the formula (ll-2-7a), is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-1-3a), optionally together with the compound of the formula (ll-2-7a), based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[0623] In another specific embodiment, the compound of the formula (ll-4-3a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-3a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 21. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 21. An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-c) and the compound of the formula (11-2), (11-3) and / or (11-4) is a compound of the formula (11-2-10), (II-3-3) and / or (II-4-4)

[0624]

[0625] preferably, a compound of the formula (II-2-10a), (I l-3-3a) and / or (ll-4-4a)

[0626] (II-2-10a),

[0627] (ll-3-3a),

[0628]

[0629] (ll-4-4a).

[0630]

[0631] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-5, 16, 17, 19, 22, 25, 27-30, 32, 34-37, 39-45, 49, 51, 53-65, 67-69, 102, 104, 107, 110, 134, 137 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-5, 16, 17, 19, 22, 25, 27-30, 32, 34-37, 39-45, 49, 51, 53-65, 67-69, 102, 104, 107, 110, 134, 137 and 139.

[0632] In a specific embodiment, the compound of the formula (ll-3-3a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-3a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 5, 16, 17, 19, 22, 25, 28, 29, 32, 35, 36, 39, 41 to 45, 49, 51, 54 to 62, 64, 65, 67 to 69, 102, 104, 107, 110 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 5, 16, 17, 19, 22, 25, 28, 29, 32, 35, 36, 39, 41 to 45, 49, 51, 54 to 62, 64, 65, 67 to 69, 102, 104, 107, 110 and 139.

[0633] In another specific embodiment, the compound of the formula (11-2-10a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-2-10a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 134. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 134.

[0634] In another specific embodiment, the compound of the formula (ll-4-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-4a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 27, 30, 34, 37, 40, 53, 63 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 27, 30, 34, 37, 40, 53, 63 and 137.

[0635] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-d) and the compound of the formula (11-1), (II-2), (II-3) and / or (II-4) is a compound of the formula (11-1-2), (II-2-4), (II-2-5), (II-A), (11-4-1) and / or (II-4-2)

[0636] (l-d),

[0637] (II-1-2),

[0638]

[0639] (11-2-4),

[0640] (II-2-5),

[0641] (II-A),

[0642] (II-4-1),

[0643] (II-4-2);

[0644]

[0645] preferably, a compound of the formula (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (II-4-1b) and / or (II-4-2b)

[0646] (ll-1-2a),

[0647]

[0648] (II-2-4a),

[0649] (II-2-5),

[0650] (II-Aa),

[0651] (II-4-1b),

[0652] (II-4-2b).

[0653]

[0654] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8, 10, 14, 16-20, 22, 25, 27-37, 39, 40-46, 49, 50-65, 67-69, 102-113, 117, 118, 121, 123-127, 129-131, 134 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8, 10, 14, 16-20, 22, 25, 27-37, 39, 40-46, 49, 50-65, 67-69, 102-113, 117, 118, 121, 123-127, 129-131, 134 and 137-139.

[0655] In a specific embodiment, the compound of the formula (II-Aa), (ll-4-2b) and / or (11-4-1 b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-Aa), (ll-4-2b) and / or (11-4-1 b) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28 to 37, 39 to 45, 49 to 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134 and 137 to 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28 to 37, 39 to 45, 49 to 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134 and 137 to 139.

[0656] In a more specific embodiment, the compound of the formula (ll-Aa) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Aa) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16, 17 to 19, 22, 25, 28 to 33, 35, 36, 39 to 45, 49 to 52, 54 to 62, 64, 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16, 17 to 19, 22, 25, 28 to 33, 35, 36, 39 to 45, 49 to 52, 54 to 62, 64, 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134, 138 and 139.

[0657] In another specific embodiment, the compound of the formula (ll-2-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II- 2-4a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 27. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 27.

[0658] In another specific embodiment, the compound of the formula (II-2-5) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-5) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46.

[0659] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-e) and the compound of the formula (11-1), (II-2), (II-3) and / or (II-4) is a compound of the formula (11-1-1), (11-2-1), (II-2-2), (II-2-3), (11-3-1) (11-4-1) and / or (II-4-2)

[0660] (l-e),

[0661] (II-1-1),

[0662]

[0663] (11-2-1),

[0664] OH (H-2-2),

[0665] (II-2-3),

[0666] (II-3-1),

[0667] (II-4-1),

[0668] (II-4-2);

[0669]

[0670] preferably, a compound of the formula (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (II-4-2a) (II-1-1a),

[0671] (II-2-1),

[0672] (ll-2-2a),

[0673] (II-2-3a),

[0674] (II-3-1a),

[0675] (II-4-1a),

[0676] (II-4-2a).

[0677]

[0678] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-6, 14, 16-20, 22, 24-30, 32, 34-37, 39, 41, 43-51, 55-73, 80-89, 92-94, 96-102, 104, 107, 113, 120, 126, 127, 132 and 137-141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-6, 14, 16-20, 22, 24-30, 32, 34-37, 39, 41, 43-51, 55-73, 80-89, 92-94, 96-102, 104, 107, 113, 120, 126, 127, 132 and 137-141.

[0679] In a specific embodiment, the compound of the formula (ll-3-1a) and / or (ll-4-1a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-3-1 a) and / or (11-4-1 a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 30, 32, 34 to 37, 39, 41, 43 to 45, 47 to 51, 55 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 137 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 30, 32, 34 to 37, 39, 41, 43 to 45, 47 to 51, 55 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 137 to 140.

[0680] In a more specific embodiment, the compound of the formula (11-3-1 a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-3-1a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 26, 28, 29, 32, 35, 36, 39, 41, 43 to 45, 47 to 51, 55 to 62, 64 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 138 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 26, 28, 29, 32, 35, 36, 39, 41, 43 to 45, 47 to 51, 55 to 62, 64 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 138 to 140.

[0681] In a more specific embodiment, the compound of the formula (11-4-1 a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-1a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 27, 30, 34, 37, 63 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 27, 30, 34, 37, 63 and 137.

[0682] In another specific embodiment, the compound of the formula (11-2-1) and / or (ll-2-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-2-1) and / or (II-2-2a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 46, 92 to 94, 96 to 101 and 141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 46, 92 to 94, 96 to 101 and 141.

[0683] In a more specific embodiment, the compound of the formula (ll-2-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-2a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 100. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 100.

[0684] In another specific embodiment, the compound of the formula (11-2-1) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-1) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (11-1 -1 a), (II-2-1), (II-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 46, 92 to 94, 96 to 99, 101 and 141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 46, 92 to 94, 96 to 99, 101 and 141.

[0685] In another specific embodiment, the compound of the formula (ll-2-3a) and / or (11-1 -1a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (I l-2-3a) and / or (11-1 -1 a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[0686] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-f) and the compound of the formula (II-2) is a compound of the formula (II-2-4) (l-f),

[0687] (II-2-4);

[0688]

[0689] preferably, a compound of the formula (ll-2-4b)

[0690] (ll-2-4b).

[0691]

[0692] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 15, 20, 21, 24, 38, 43, 48, 50, 55, 71-73, 93, 97, 100, 121 and 162-180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 15, 20, 21, 24, 38, 43, 48, 50, 55, 71-73, 93, 97, 100, 121 and 162-180.

[0693] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-g) and the compound of the formula (11-1) is a compound of the formula (ll-B) (1-g),

[0694] (II-B);

[0695]

[0696] preferably, a compound of the formula (II-Ba)

[0697] (II-Ba).

[0698]

[0699] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 8-10, 14, 17, 19, 20, 22, 25, 27-30, 32-38, 40, 42-46, 49-65, 67-69, 102, 103, 107, 118, 129, 131 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 8-10, 14, 17, 19, 20, 22, 25, 27-30, 32-38, 40, 42-46, 49-65, 67-69, 102, 103, 107, 118, 129, 131 and 137-139.

[0700] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-h) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (I l-C) and / or (II-4-5) (Il-C),

[0701] (11-4-5);

[0702]

[0703] preferably, a compound of the formula (ll-Ca) and / or (II-4-5a)

[0704] (ll-Ca),

[0705] (ll-4-5a).

[0706]

[0707] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 8, 14, 16, 17, 19, 20, 22, 25, 27-45, 49, 50, 51-65, 67-69, 102-104, 107, 109-110, 114, 116, 129 and 132-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 8, 14, 16, 17, 19, 20, 22, 25, 27-45, 49, 50, 51-65, 67-69, 102-104, 107, 109-110, 114, 116, 129 and 132-140.

[0708] In a specific embodiment, the compound of the formula (ll-Ca) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Ca) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (I I-4) (namely, (ll-Ca) and / or (II-4-5a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 5, 8, 14, 16, 17, 19, 22, 25, 28, 29 to 31, 33 to 45, 49 to 65, 67 to 69, 102 to 104, 107, 109, 110, 114, 116, 129 and 132 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 5, 8, 14, 16, 17, 19, 22, 25, 28, 29 to 31, 33 to 45, 49 to 65, 67 to 69, 102 to 104, 107, 109, 110, 114, 116, 129 and 132 to 140.

[0709] In another specific embodiment, the compound of the formula (ll-4-5a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-5a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-Ca) and / or (ll-4-5a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 20, 27 and 32. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 20, 27 and 32.

[0710] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-i) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (II-C), (II-4-5) and / or (II-4-6) (l-i),

[0711] (II-C),

[0712] (II-4-5),

[0713] (11-4-6);

[0714]

[0715] preferably, a compound of the formula (II-Cb), (II-4-5b) and / or (ll-4-6a)

[0716] (II-Cb),

[0717]

[0718] (II-4-5b),

[0719] (II-4-6a).

[0720]

[0721] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-46, 49-65, 67-69, 102-104, 106, 107-110, 112-115, 118, 119, 124-127, 129-131, 134 and 137-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-46, 49-65, 67-69, 102-104, 106, 107-110, 112-115, 118, 119, 124-127, 129-131, 134 and 137-140.

[0722] In a specific embodiment, the compound of the formula (ll-Cb) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Cb) based on the total quantified product compound (11-1), (II-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 5, 6, 8, 9, 14, 16, 17, 19, 20, 27 to 29, 31, 33, 36, 41, 43 to 46, 49, 50, 52, 53, 55, 60, 62, 64, 65, 106, 112 to 115, 119, 124 to 127, 130 and 134. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 5, 6, 8, 9, 14, 16, 17, 19, 20, 27 to 29, 31, 33, 36, 41, 43 to 46, 49, 50, 52, 53, 55, 60, 62, 64, 65, 106, 112 to 115, 119, 124 to 127, 130 and 134.

[0723] In another specific embodiment, the compound of the formula (ll-4-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-6a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1, 110, 118 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1, 110, 118 and 139.

[0724] In another specific embodiment, the compound of the formula (ll-4-5b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-5b) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 3, 10, 18, 25, 30, 34, 37, 39, 63, 107 to 109, 129, 131, 137 and 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 3, 10, 18, 25, 30, 34, 37, 39, 63, 107 to 109, 129, 131, 137 and 140.

[0725] In another specific embodiment, the compound of the formula (ll-Cb) and / or (II-4-5b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Cb) and / or (II-4-5b) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-Cb), (ll-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2, 22, 32, 35, 38, 56 to 59, 61, 67 to 69, 102 and 104. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2, 22, 32, 35, 38, 56 to 59, 61, 67 to 69, 102 and 104.

[0726] In another specific embodiment, the compound of the formula (II-Cb), (ll-4-5b) and / or (ll-4-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-Cb), (ll-4-5b) and / or (ll-4-6a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-Cb), (ll-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 4, 40, 42, 51, 54, 103 and 138. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 4, 40, 42, 51, 54, 103 and 138.

[0727] An embodiment of the inventin is characterized in that the compound of the formula (I) is a compound of the formula (l-j) and the compound of the formula (II-3) is a compound of the formula (ll-D)

[0728]

[0729] preferably, a compound of the formula (II-Da)

[0730] (II-Da).

[0731]

[0732] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 73, 74 and 173. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 73, 74 and 173.

[0733] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-k) and the compound of the formula (11-1) and / or (I I-2) is a compound of the formula (11-1-4), (11-2-12), (11-2-13) and / or (11-2-14)

[0734] (II-1-4),

[0735] (11-2-12),

[0736] (11-2-13),

[0737]

[0738] (11-2-14);

[0739]

[0740] preferably, a compound of the formula (11-1 -4a), (11-2-12a), (11-2-13) and / or (11-2-14a)

[0741] (II-1-4a),

[0742] (II-2-12a),

[0743] (11-2-13),

[0744] (II-2-14a).

[0745]

[0746] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6, 14, 20 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6, 14, 20 and 46. In a specific embodiment, the compound of the formula (ll-1-4a) and / or (ll-2-12a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-1 -4a) and / or (11-2-12a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6, 14 and 20. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6, 14 and 20.

[0747] In a more specific embodiment, the compound of the formula (ll-2-12a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-12a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6 and 14. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6 and 14.

[0748] In another specific embodiment, the compound of the formula (11-2-13) and / or (ll-2-14a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-2-13) and / or (11-2-14a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46. An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-l) and the compound of the formula (I I-3) is a compound of the formula (ll-E)

[0749]

[0750] preferably, a compound of the formula (II-Ea)

[0751] (II-Ea).

[0752]

[0753] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8, 10, 14, 16, 17, 19, 22, 25, 27-45, 49-65, 67-69, 102-110, 114, 118, 124, 126, 129, 131, 134 and 137-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 -6, 8, 10, 14, 16, 17, 19, 22, 25, 27-45, 49-65, 67-69, 102-110, 114, 118, 124, 126, 129, 131, 134 and 137-140. An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-m) and the compound of the formula (11-1) and / or (II-2) is a compound of the formula (11-1-5), (11-2-15) and / or (11-2-16)

[0754] (l-m),

[0755] (II-1-5),

[0756] (II-2-15),

[0757] (11-2-16);

[0758]

[0759] preferably, a compound of the formula (II-1-5a), (II-2-15a) and / or (II-2-16)

[0760] (ll-1-5a),

[0761] (11-2-15a),

[0762] (11-2-16).

[0763]

[0764] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 20 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 20 and 46.

[0765] In a specific embodiment, the compound of the formula (ll-1-5a) and / or (ll-2-15a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-1 -5a) and / or (11-2-15a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-5a), (II-2-15a) and / or (II-2-16)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[0766] In another specific embodiment, the compound of the formula (11-2-16) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-16) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-5a), (II-2-15a) and / or (II-2-16)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46. An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-n) and the compound of the formula (II-3) is a compound of the formula (II-3-5)

[0767] (l-n),

[0768] (II-3-5);

[0769]

[0770] preferably, a compound of the formula (II-3-5a)

[0771] (II-3-5a).

[0772]

[0773] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 14, 17, 19, 20, 22, 27-29, 32, 35, 36, 43-46, 49-51, 54-60, 62, 65, 67-69, 102, 107, 129, 131 and 138. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 14, 17, 19, 20, 22, 27-29, 32, 35, 36, 43-46, 49-51, 54-60, 62, 65, 67-69, 102, 107, 129, 131 and 138.

[0774] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-o) and the compound of the formula (11-1) is a compound of the formula (ll-F) (l-o),

[0775] (II-F);

[0776]

[0777] preferably, the compound of the formula (l-o) is a compound of the formula (l-oa) and the compound of the formula (II-F) is a compound of the formula (II-Fa)

[0778] (II-Fa);

[0779]

[0780] also preferably, the compound of the formula (l-o) is a compound of the formula (l-ob) and the compound of the formula (II-F) is a compound of the formula (II-Fb)

[0781] (l-ob),

[0782] (II-Fb).

[0783]

[0784] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, 2024P00180WO

[0785] the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 14, 20 and 36. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 14, 20 and 36.

[0786] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-p) and the compound of the formula (II-2) and / or (II-3) is a compound of the formula (11-2-11) and / or (II-3-4)

[0787] (11-2-11),

[0788] (II-3-4);

[0789]

[0790] preferably, a compound of the formula (11-2-11a) and / or (ll-3-4a)

[0791] (II-2-11a),

[0792] (II-3-4a).

[0793]

[0794] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-36, 38-46, 49-63, 65, 67-69, 102-116, 118, 119, 121-129 and 131-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-36, 38-46, 49-63, 65, 67-69, 102-116, 118, 119, 121-129 and 131-140.

[0795] In a specific embodiment, the compound of the formula (II-3-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-3-4a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-11a) and / or (II-3-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28, 29, 31 to 36, 38 to 46, 49 to 62, 65, 67 to 69, 102 to 108, 110 to 116, 118, 119, 122, 124 to 127, 129, 131 to 133, 135 and 138 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28, 29, 31to 36, 38 to 46, 49 to 62, 65, 67 to 69, 102 to 108, 110 to 116, 118, 119, 122, 124 to 127, 129, 131 to 133, 135 and 138 to 140.

[0796] In another specific embodiment, the compound of the formula (11-2-11a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-2-11a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-2-11a) and / or (II-3-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 9, 27, 30, 63, 109, 121, 123, 125, 128, 134, 136 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 9, 27, 30, 63, 109, 121, 123, 125, 128, 134, 136 and 137.

[0797] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-q) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (II-3-6) and / or (II-4-7)

[0798] (II-3-6),

[0799] (II-4-7);

[0800]

[0801] preferably, a compound of the formula (ll-3-6a) and / or (II-4-7a)

[0802] (II-3-6a),

[0803] (II-4-7a).

[0804]

[0805] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16, 17-20, 22, 25, 27-46, 49-65, 67-69 and 102-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16, 17-20, 22, 25, 27-46, 49-65, 67-69 and 102-140.

[0806] In a specific embodiment, the compound of the formula (I l-3-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-6a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-3-6a) and / or (II-4-7a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 14, 16 to 19, 22, 25, 28, 29, 32, 35, 36, 39 to 45, 49, 51, 52, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 112, 114, 122, 124, 125, 128, 136, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 14, 16 to 19, 22, 25, 28, 29, 32, 35, 36, 39 to 45, 49, 51, 52, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 112, 114, 122, 124, 125, 128, 136, 138 and 139.

[0807] In another specific embodiment, the compound of the formula (ll-4-7a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-7a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-3-6a) and / or (II-4-7a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 8 to 10, 20, 27, 30, 31, 33, 34, 37, 38, 46, 50, 53, 63, 105, 106, 108, 109, 111, 113, 115 to 121, 123, 126, 127, 129 to 135, 137 and 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 8 to 10, 20, 27, 30, 31, 33, 34, 37, 38, 46, 50, 53, 63, 105, 106, 108, 109, 111, 113, 115 to 121, 123, 126, 127, 129 to 135, 137 and 140.

[0808] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-r) and the compound of the formula (II-3) is a compound of the formula (II-3-7)

[0809] (II-3-7);

[0810]

[0811] preferably, a compound of the formula (II-3-7a)

[0812] (II-3-7a).

[0813]

[0814] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 29, 48, 69, 71, 73, 74, 75, 76, 77 and 78. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 29, 48, 69, 71, 73, 74, 75, 76, 77 and 78.

[0815] An embodiment of the invention is characterized in that the compound of the formula (I) is a compound of the formula (l-s) and the compound of the formula (11-3) is a compound of the formula (11-3-8)

[0816] (II-3-8);

[0817]

[0818] preferably, a compound of the formula (ll-3-8a)

[0819] (ll-3-8a).

[0820]

[0821] In said above embodiment of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined herein above. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 79. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 79.

[0822] Further aspects of the invention

[0823] A further aspect of the invention provides a method for preparing a compound of the formula (II-2-17)

[0824]

[0825] characterized in that the method comprises contacting a compound of the formula (l-a)

[0826] (l-a),

[0827]

[0828] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-2-17).

[0829] Preferably, the compound of the formula (11-2-17) is a compound of the formula (11-2-17a)

[0830] (11-2-17a).

[0831]

[0832] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 38. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 38.

[0833] A further aspect of the invention provides a method for preparing a compound of the formula (II- 1-3), (II-2-7), (II-2-8), (II-2-9), (II-3-2) and / or (II-4-3)

[0834] (II-1-3),

[0835] (II-2-7),

[0836] (II-2-8),

[0837] (II-2-9),

[0838] (II-3-2),

[0839]

[0840] (11-4-3),

[0841]

[0842] characterized in that the method comprises contacting a compound of the formula (l-b)

[0843] (l-b),

[0844]

[0845] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-1-3), (II-2-7), (II-2-8), (II-2-9), (II-3-2) and / or (II-4-3).

[0846] Preferably, the compound of the formula (II-1-3), (II-2-7), (II-2-8), (II-2-9), (II-3-2) and / or (II-4-3) is a compound of the formula (11-1 -3a), (ll-2-7a), (II-2-8), (II-2-9), (ll-3-2a) and / or (ll-4-3a)

[0847] (ll-1-3a),

[0848] (ll-2-7a),

[0849] (ll-2-8a),

[0850] (II-2-9),

[0851]

[0852] (II-3-2a),

[0853] (II-4-3a).

[0854]

[0855] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-8, 11-17, 19-22, 25, 28, 29, 32, 35, 36, 38, 39, 41 -46, 49-51, 54-62, 64, 65, 67-69, 102-104, 107, 110 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-8, 11-17, 19-22, 25, 28, 29, 32, 35, 36, 38, 39, 41-46, 49-51, 54-62, 64, 65, 67-69, 102-104, 107, 110 and 137-139.

[0856] In a specific embodiment, the compound of the formula (ll-3-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-2a) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (I I-4) (namely, (11-1 -3a), (I l-2-7a), (II-2-8a), (II-2-9), (ll-3-2a) and / or (I l-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 8, 14 to 17, 19, 22, 25, 28, 29, 32, 35, 36, 38, 39, 41 to 45, 49 to 51, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 137, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 8, 14 to 17, 19, 22, 25, 28, 29, 32, 35, 36, 38, 39, 41 to 45, 49 to 51, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 137, 138 and 139.

[0857] In another specific embodiment, the compound of the formula (II-2-9) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-9) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (II-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 7 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 7 and 46.

[0858] In another specific embodiment, the compound of the formula (ll-2-8a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-8a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 11 to 13. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 11 to 13.

[0859] In another specific embodiment, the compound of the formula (11-1 -3a), optionally together with the compound of the formula (ll-2-7a), is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-1-3a), optionally together with the compound of the formula (ll-2-7a), based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[0860] In another specific embodiment, the compound of the formula (ll-4-3a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-3a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1 -3a), (ll-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (ll-4-3a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 21. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 21.

[0861] A further aspect of the invention provides a method for preparing a compound of the formula (II-2-10), (II-3-3) and / or (I I-4-4)

[0862]

[0863] characterized in that the method comprises contacting a compound of the formula (l-c)

[0864]

[0865] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-2-10), (II-3-3) and / or (II-4-4).

[0866] Preferably, the compound of the formula (11-2-10), (II-3-3) and / or (II-4-4) is a compound of the formula (11-2-10a), (I l-3-3a) and / or (ll-4-4a)

[0867] (II-2-10a),

[0868] (II-3-3a),

[0869] (II-4-4a).

[0870]

[0871] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-5, 16, 17, 19, 22, 25, 27-30, 32, 34-37, 39-45, 49, 51, 53- 65, 67-69, 102, 104, 107, 110, 134, 137 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-5, 16, 17, 19, 22, 25, 27-30, 32, 34-37, 39-45, 49, 51, 53-65, 67-69, 102, 104, 107, 110, 134, 137 and 139.

[0872] In a specific embodiment, the compound of the formula (ll-3-3a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-3a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 5, 16, 17, 19, 22, 25, 28, 29, 32, 35, 36, 39, 41 to 45, 49, 51, 54 to 62, 64, 65, 67 to 69, 102, 104, 107, 110 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 5, 16, 17, 19, 22, 25, 28, 29, 32, 35, 36, 39, 41 to 45, 49, 51, 54 to 62, 64, 65, 67 to 69, 102, 104, 107, 110 and 139.

[0873] In another specific embodiment, the compound of the formula (11-2-10a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-2-10a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 134. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 134.

[0874] In another specific embodiment, the compound of the formula (ll-4-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II-4-4a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (11-2-10a), (ll-3-3a) and / or (ll-4-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 5%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 27, 30, 34, 37, 40, 53, 63 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 27, 30, 34, 37, 40, 53, 63 and 137.

[0875] A further aspect of the invention provides a method for preparing a compound of the formula (II--2), (II-2-4), (II-2-5), (II-A), (11-4-1) and / or (II-4-2)

[0876] (II-1-2),

[0877] (II-2-4),

[0878] (II-2-5),

[0879] (II-A),

[0880] (II-4-1),

[0881]

[0882] (II-4-2);

[0883]

[0884] characterized in that the method comprises contacting a compound of the formula (l-d)

[0885] (l-d),

[0886]

[0887] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-1-2), (II-2-4), (II-2-5), (II-A), (11-4-1) and / or (II-4-2).

[0888] Preferably, the compound of the formula (11-1-2), (II-2-4), (II-2-5), (II-A), (11-4-1) and / or (II-4-2) is a compound of the formula (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)

[0889] (ll-1-2a),

[0890]

[0891] (II-2-4a),

[0892] (II-2-5),

[0893]

[0894] (II-Aa),

[0895] (II-4-1b),

[0896] (II-4-2b).

[0897]

[0898] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8, 10, 14, 16-20, 22, 25, 27-37, 39, 40-46, 49, 50-65, 67-69, 102-113, 117, 118, 121, 123-127, 129-131, 134 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8, 10, 14, 16-20, 22, 25, 27-37, 39, 40-46, 49, 50-65, 67-69, 102-113, 117, 118, 121, 123-127, 129-131, 134 and 137-139.

[0899] In a specific embodiment, the compound of the formula (II-Aa), (ll-4-2b) and / or (11-4-1 b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-Aa), (ll-4-2b) and / or (11-4-1 b) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28 to 37, 39 to 45, 49 to 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134 and 137 to 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28 to 37, 39 to 45, 49 to 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134 and 137 to 139.

[0900] In a more specific embodiment, the compound of the formula (ll-Aa) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Aa) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16, 17 to 19, 22, 25, 28 to 33, 35, 36, 39 to 45, 49 to 52, 54 to 62, 64, 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16, 17 to 19, 22, 25, 28 to 33, 35, 36, 39 to 45, 49 to 52, 54 to 62, 64, 65, 67 to 69, 102 to 113, 117, 118, 121, 123 to 127, 129 to 131, 134, 138 and 139.

[0901] In another specific embodiment, the compound of the formula (ll-2-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-4a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 27. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 27. In another specific embodiment, the compound of the formula (11-2-5) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-5) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (II-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46.

[0902] A further aspect of the invention provides a method for preparing a compound of the formula (II-1-1), (11-2-1), (II-2-2), (II-2-3), (11-3-1) (11-4-1) and / or (II-4-2)

[0903] (II-1-1),

[0904] (II-2-1),

[0905] (II-2-2),

[0906] (II-2-3),

[0907]

[0908] (11-3-1),

[0909] (II-4-1),

[0910] (II-4-2);

[0911]

[0912] characterized in that the method comprises contacting a compound of the formula (l-e)

[0913]

[0914] ' OH with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-1-1), (11-2-1), (II-2-2), (II-2-3), (11-3-1) (11-4-1) and / or (II-4-2).

[0915] Preferably, the compound of the formula (11-1-1), (11-2-1), (II-2-2), (II-2-3), (11-3-1) (11-4-1) and / or (II-4-2) is a compound of the formula (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (II-4-2a)

[0916] (II-1-1a),

[0917]

[0918] 2024P00180WO

[0919] (11-2-1),

[0920] (ll-2-2a),

[0921] (II-2-3a),

[0922] (II-3-1a),

[0923] (II-4-1a),

[0924] (II-4-2a).

[0925]

[0926] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-6, 14, 16-20, 22, 24-30, 32, 34-37, 39, 41, 43-51, 55-73, 80-89, 92-94, 96-102, 104, 107, 113, 120, 126, 127, 132 and 137-141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-6, 14, 16-20, 22, 24-30, 32, 34-37, 39, 41, 43-51, 55-73, 80-89, 92-94, 96-102, 104, 107, 113, 120, 126, 127, 132 and 137-141.

[0927] In a specific embodiment, the compound of the formula (ll-3-1a) and / or (ll-4-1a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-3-1 a) and / or (11-4-1 a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 30, 32, 34 to 37, 39, 41, 43 to 45, 47 to 51, 55 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 137 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 30, 32, 34 to 37, 39, 41, 43 to 45, 47 to 51, 55 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 137 to 140.

[0928] In a more specific embodiment, the compound of the formula (11-3-1 a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-3-1a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 26, 28, 29, 32, 35, 36, 39, 41, 43 to 45, 47 to 51, 55 to 62, 64 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 138 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 6, 14, 16 to 19, 22, 24 to 26, 28, 29, 32, 35, 36, 39, 41, 43 to 45, 47 to 51, 55 to 62, 64 to 73, 80 to 89, 102, 104, 107, 113, 120, 126, 127, 132 and 138 to 140.

[0929] In a more specific embodiment, the compound of the formula (11-4-1 a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-1a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 27, 30, 34, 37, 63 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 27, 30, 34, 37, 63 and 137.

[0930] In another specific embodiment, the compound of the formula (11-2-1) and / or (ll-2-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-2-1) and / or (II-2-2a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 46, 92 to 94, 96 to 101 and 141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 46, 92 to 94, 96 to 101 and 141.

[0931] In a more specific embodiment, the compound of the formula (ll-2-2a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-2a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (11-1-1 a), (11-2-1), (ll-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 100. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 100.

[0932] In another specific embodiment, the compound of the formula (11-2-1) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-1) based on the total quantified product compound (11-1), (I I-2), (II-3) and / or (II-4) (namely, (11-1 -1 a), (II-2-1), (II-2-2a), (ll-2-3a), (11-3-1 a) (11-4-1 a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 46, 92 to 94, 96 to 99, 101 and 141. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 46, 92 to 94, 96 to 99, 101 and 141.

[0933] In another specific embodiment, the compound of the formula (ll-2-3a) and / or (11-1 -1a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (I l-2-3a) and / or (11-1 -1 a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-1-1a), (11-2-1), (ll-2-2a), (ll-2-3a), (ll-3-1a) (ll-4-1a) and / or (ll-4-2a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[0934] A further aspect of the invention provides a method for preparing a compound of the formula (II-2-4)

[0935] (II-2-4),

[0936]

[0937] characterized in that the method comprises contacting a compound of the formula (l-f)

[0938]

[0939] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-2-4).

[0940] Preferably, the compound of the formula (II-2-4) is a compound of the formula (ll-2-4b)

[0941] (ll-2-4b).

[0942]

[0943] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 15, 20, 21, 24, 38, 43, 48, 50, 55, 71-73, 93, 97, 100, 121 and 162-180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 15, 20, 21, 24, 38, 43, 48, 50, 55, 71-73, 93, 97, 100, 121 and 162-180.

[0944] A further aspect of the invention provides a method for preparing a compound of the formula (II-B) (ll-B),

[0945]

[0946] characterized in that the method comprises contacting a compound of the formula

[0947] (i-g).

[0948]

[0949] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (ll-B).

[0950] Preferably, the compound of the formula (ll-B) is a compound of the formula (II-Ba)

[0951] (II-Ba).

[0952]

[0953] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 8-10, 14, 17, 19, 20, 22, 25, 27-30, 32-38, 40, 42-46, 49-65, 67-69, 102, 103, 107, 118, 129, 131 and 137-139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 8-10, 14, 17, 19, 20, 22, 25, 27-30, 32-38, 40, 42-46, 49-65, 67-69, 102, 103, 107, 118, 129, 131 and 137-139. 2024P00180WO

[0954] A further aspect of the invention provides a method for preparing a compound of the formula (II- C) and / or (11-4-5)

[0955] (II-C),

[0956] (II-4-5);

[0957]

[0958] characterized in that the method comprises contacting a compound of the formula (l-h)

[0959]

[0960] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (ll-C) and / or (11-4-5).

[0961] Preferably, the compound of the formula (ll-C) and / or (II-4-5) is a compound of the formula (Il- Ca) and / or (ll-4-5a)

[0962] (ll-Ca),

[0963] (ll-4-5a).

[0964]

[0965] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 8, 14, 16, 17, 19, 20, 22, 25, 27-45, 49, 50, 51-65, 67-69, 102-104, 107, 109-110, 114, 116, 129 and 132-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 8, 14, 16, 17, 19, 20, 22, 25, 27-45, 49, 50, 51-65, 67-69, 102-104, 107, 109-110, 114, 116, 129 and 132-140.

[0966] In a specific embodiment, the compound of the formula (ll-Ca) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Ca) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (I I-4) (namely, (ll-Ca) and / or (II-4-5a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2 to 5, 8, 14, 16, 17, 19, 22, 25, 28, 29 to 31, 33 to 45, 49 to 65, 67 to 69, 102 to 104, 107, 109, 110, 114, 116, 129 and 132 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2 to 5, 8, 14, 16, 17, 19, 22, 25, 28, 29 to 31, 33 to 45, 49 to 65, 67 to 69, 102 to 104, 107, 109, 110, 114, 116, 129 and 132 to 140.

[0967] In another specific embodiment, the compound of the formula (ll-4-5a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-5a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (ll-Ca) and / or (ll-4-5a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 2024P00180WO

[0968] 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 20, 27 and 32. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 20, 27 and 32.

[0969] A further aspect of the invention provides a method for preparing a compound of the formula (II-C), (11-4-5) and / or (11-4-6)

[0970] (II-C),

[0971] (II-4-5),

[0972] (II-4-6);

[0973]

[0974] characterized in that the method comprises contacting a compound of the formula (l-i)

[0975]

[0976] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-C), (11-4-5) and / or (11-4-6).

[0977] Preferably, the compound of the formula (II-C), (11-4-5) and / or (11-4-6) is a compound of the formula (II-Cb), (ll-4-5b) and / or (ll-4-6a) (II-Cb),

[0978] (II-4-5b),

[0979] (II-4-6a).

[0980]

[0981] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-46, 49-65, 67-69, 102-104, 106, 107-110, 112-115, 118, 119, 124-127, 129-131, 134 and 137-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-46, 49-65, 67-69, 102-104, 106, 107-110, 112-115, 118, 119, 124-127, 129-131, 134 and 137-140.

[0982] In a specific embodiment, the compound of the formula (ll-Cb) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Cb) based on the total quantified product compound (11-1), (II-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 5, 6, 8, 9, 14, 16, 17, 19, 20, 27 to 29, 31, 33, 36, 41, 43 to 46, 49, 50, 52, 53, 55, 60, 62, 64, 65, 106, 112 to 115, 119, 124 to 127, 130 and 134. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 5, 6, 8, 9, 14, 16, 17, 19, 20, 27 to 29, 31, 33, 36, 41, 43 to 46, 49, 50, 52, 53, 55, 60, 62, 64, 65, 106, 112 to 115, 119, 124 to 127, 130 and 134.

[0983] In another specific embodiment, the compound of the formula (ll-4-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-6a) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1, 110, 118 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1, 110, 118 and 139.

[0984] In another specific embodiment, the compound of the formula (ll-4-5b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-5b) based on the total quantified product compound (11-1), (I I-2), (I I-3) and / or (I I-4) (namely, (II-Cb), (II-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 3, 10, 18, 25, 30, 34, 37, 39, 63, 107 to 109, 129, 131, 137 and 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 3, 10, 18, 25, 30, 34, 37, 39, 63, 107 to 109, 129, 131, 137 and 140.

[0985] In another specific embodiment, the compound of the formula (ll-Cb) and / or (II-4-5b) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (ll-Cb) and / or (II-4-5b) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-Cb), (ll-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2, 22, 32, 35, 38, 56 to 59, 61, 67 to 69, 102 and 104. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2, 22, 32, 35, 38, 56 to 59, 61, 67 to 69, 102 and 104.

[0986] In another specific embodiment, the compound of the formula (II-Cb), (ll-4-5b) and / or (ll-4-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-Cb), (ll-4-5b) and / or (ll-4-6a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-Cb), (ll-4-5b) and / or (ll-4-6a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 4, 40, 42, 51, 54, 103 and 138. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 4, 40, 42, 51, 54, 103 and 138.

[0987] A further aspect of the invention provides a method for preparing a compound of the formula (II-D)

[0988]

[0989] characterized in that the method comprises contacting a compound of the formula (l-j)

[0990]

[0991] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (ll-D).

[0992] Preferably, the compound of the formula (ll-D) is a compound of the formula (II-Da)

[0993] (II-Da).

[0994]

[0995] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 73, 74, 173. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 73, 74, 173.

[0996] A further aspect of the invention provides a method for preparing a compound of the formula (II-1-4), (11-2-12), (11-2-13) and / or (11-2-14)

[0997] (II-1-4),

[0998] (11-2-12),

[0999]

[1000] (11-2-13),

[1001] (11-2-14);

[1002]

[1003] characterized in that the method comprises contacting a compound of the formula (l-k)

[1004]

[1005] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-1-4), (11-2-12), (11-2-13) and / or (11-2-14).

[1006] Preferably, the compound of the formula (11-1-4), (11-2-12), (11-2-13) and / or (11-2-14) is a compound of the formula (II-1-4a), (II-2-12a), (11-2-13) and / or (11-2-14a)

[1007] (II-1-4a),

[1008] (II-2-12a),

[1009]

[1010] (11-2-13),

[1011] o

[1012] (II-2-14a).

[1013]

[1014] o In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6, 14, 20 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6, 14, 20 and 46.

[1015] In a specific embodiment, the compound of the formula (ll-1-4a) and / or (ll-2-12a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-1 -4a) and / or (11-2-12a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6, 14 and 20. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6, 14 and 20.

[1016] In a more specific embodiment, the compound of the formula (ll-2-12a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-12a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 6 and 14. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 6 and 14. In another specific embodiment, the compound of the formula (11-2-13) and / or (ll-2-14a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-2-13) and / or (11-2-14a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-4a), (II-2-12a), (II-2-13) and / or (II-2-14a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46.

[1017] A further aspect of the invention provides a method for preparing a compound of the formula (II-E)

[1018]

[1019] characterized in that the method comprises contacting a compound of the formula (l-l)

[1020]

[1021] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (ll-E).

[1022] Preferably, the compound of the formula (ll-E) is a compound of the formula (II-Ea)

[1023] (II-Ea).

[1024]

[1025] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8, 10, 14, 16, 17, 19, 22, 25, 27-45, 49-65, 67-69, 102-110, 114, 118, 124, 126, 129, 131, 134 and 137-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8, 10, 14, 16, 17, 19, 22, 25, 27-45, 49-65, 67-69, 102-110, 114, 118, 124, 126, 129, 131, 134 and 137-140.

[1026] A further aspect of the invention provides a method for preparing a compound of the formula (II- 1-5), (11-2-15) and / or (11-2-16)

[1027] (II-1-5),

[1028] (II-2-15),

[1029]

[1030] characterized in that the method comprises contacting a compound of the formula (l-m)

[1031] (l-m),

[1032]

[1033] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-1-5), (11-2-15) and / or (11-2-16). Preferably, the compound of the formula (11-1-5), (11-2-15) and / or (11-2-16) is a compound of the formula (ll-1-5a), (ll-2-15a) and / or (11-2-16)

[1034] (ll-1-5a),

[1035]

[1036] (11-2-15a),

[1037] (11-2-16).

[1038]

[1039] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 20 and 46. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 20 and 46.

[1040] In a specific embodiment, the compound of the formula (ll-1-5a) and / or (ll-2-15a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (11-1 -5a) and / or (11-2-15a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-5a), (II-2-15a) and / or (II-2-16)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 20. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 20.

[1041] In another specific embodiment, the compound of the formula (11-2-16) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-2-16) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-1-5a), (II-2-15a) and / or (II-2-16)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 46. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 46.

[1042] A further aspect of the invention provides a method for preparing a compound of the formula (II-3-5)

[1043] (II-3-5),

[1044]

[1045] characterized in that the method comprises contacting a compound of the formula (l-n)

[1046] (l-n),

[1047]

[1048] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-3-5).

[1049] Preferably, the compound of the formula (II-3-5) is a compound of the formula (II-3-5a)

[1050] (II-3-5a).

[1051]

[1052] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 2-5, 14, 17, 19, 20, 22, 27-29, 32, 35, 36, 43-46, 49-51, 54-60, 62, 65, 67-69, 102, 107, 129, 131 and 138. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 2-5, 14, 17, 19, 20, 22, 27-29, 32, 35, 36, 43-46, 49-51, 54-60, 62, 65, 67-69, 102, 107, 129, 131 and 138.

[1053] A further aspect of the invention provides a method for preparing a compound of the formula (II-F)

[1054]

[1055] characterized in that the method comprises contacting a compound of the formula (l-o)

[1056]

[1057] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (ll-F).

[1058] Preferably, the compound of the formula (l-o) is a compound of the formula (l-oa) and the compound of the formula (II-F) is a compound of the formula (II-Fa)

[1059] (l-oa),

[1060] (ll-Fa).

[1061]

[1062] Also preferably, the compound of the formula (l-o) is a compound of the formula (l-ob) and the compound of the formula (II-F) is a compound of the formula (II-Fb)

[1063] (l-ob),

[1064] (II-Fb).

[1065]

[1066] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 14, 20 and 36. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 14, 20 and 36.

[1067] A further aspect of the invention provides a method for preparing a compound of the formula (II-2-11) and / or (I I-3-4)

[1068] (11-2-11),

[1069] (II-3-4);

[1070]

[1071] characterized in that the method comprises contacting a compound of the formula (l-p)

[1072] (I-p),

[1073]

[1074] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-2-11) and / or (II-3-4).

[1075] Preferably, the compound of the formula (11-2-11) and / or (II-3-4) is a compound of the formula (11-2-11a) and / or (ll-3-4a)

[1076] (II-2-11a),

[1077] (II-3-4a).

[1078]

[1079] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-36, 38-46, 49-63, 65, 67-69, 102-116, 118, 119, 121-129 and 131-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16-20, 22, 25, 27-36, 38-46, 49-63, 65, 67-69, 102-116, 118, 119, 121-129 and 131-140. In a specific embodiment, the compound of the formula (ll-3-4a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-3-4a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (11-2-11a) and / or (II-3-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28, 29, 31to 36, 38 to 46, 49 to 62, 65, 67 to 69, 102 to 108, 110 to 116, 118, 119, 122, 124 to 127, 129, 131 to 133, 135 and 138 to 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 8, 10, 14, 16 to 20, 22, 25, 28, 29, 31to 36, 38 to 46, 49 to 62, 65, 67 to 69, 102 to 108, 110 to 116, 118, 119, 122, 124 to 127, 129, 131 to 133, 135 and 138 to 140.

[1080] In another specific embodiment, the compound of the formula (11-2-11a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound ofthe formula (II- 2-11a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-2-11a) and / or (II-3-4a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 9, 27, 30, 63, 109, 121, 123, 125, 128, 134, 136 and 137. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 9, 27, 30, 63, 109, 121, 123, 125, 128, 134, 136 and 137.

[1081] A further aspect ofthe invention provides a method for preparing a compound ofthe formula (II- 3-6) and / or (I I-4-7)

[1082] (II-3-6),

[1083]

[1084] (11-4-7);

[1085]

[1086] characterized in that the method comprises contacting a compound of the formula (l-q)

[1087] (l-q),

[1088]

[1089] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (11-3-6) and / or (11-4-7).

[1090] Preferably, the compound of the formula (11-3-6) and / or (11-4-7) is a compound of the formula (II-3-6a) and / or (II-4-7a)

[1091] (II-3-6a),

[1092] (II-4-7a).

[1093]

[1094] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1-6, 8-10, 14, 16, 17-20, 22, 25, 27-46, 49-65, 67-69 and 102-140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1-6, 8-10, 14, 16, 17-20, 22, 25, 27-46, 49-65, 67-69 and 102-140.

[1095] In a specific embodiment, the compound of the formula (ll-3-6a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) ofthe compound of the formula (ll-3-6a) based on the total quantified product compound (11-1), (II-2), (II-3) and / or (II-4) (namely, (II-3-6a) and / or (II-4-7a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 6, 14, 16 to 19, 22, 25, 28, 29, 32, 35, 36, 39 to 45, 49, 51, 52, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 112, 114, 122, 124, 125, 128, 136, 138 and 139. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 6, 14, 16 to 19, 22, 25, 28, 29, 32, 35, 36, 39 to 45, 49, 51, 52, 54 to 62, 64, 65, 67 to 69, 102 to 104, 107, 110, 112, 114, 122, 124, 125, 128, 136, 138 and 139.

[1096] In another specific embodiment, the compound of the formula (ll-4-7a) is the main product as defined in the Section “Definitions”. In particular, the relative ratio (%) of the compound of the formula (II-4-7a) based on the total quantified product compound (II-1), (II-2), (II-3) and / or (II-4) (namely, (II-3-6a) and / or (II-4-7a)) is in the range of 51 to 100%, particularly in the range of 60 to 100%, particularly in the range of 70 to 100%, particularly in the range of 75 to 100%, particularly in the range of 80 to 100%, particularly in the range of 85 to 100%, particularly in the range of 90 to 100%, particularly in the range of 95 to 100%; more in particular, in the range of 96 to 100%, particularly in the range of 97 to 100%, particularly in the range of 98 to 100%, particularly in the range of 99 to 100%, particularly in the range of 99,5 to 100%, particularly in the range of 99,8 to 100%. In said embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 8 to 10, 20, 27, 30, 31, 33, 34, 37, 38, 46, 50, 53, 63, 105, 106, 108, 109, 111, 113, 115 to 121, 123, 126, 127, 129 to 135, 137 and 140. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 8 to 10, 20, 27, 30, 31, 33, 34, 37, 38, 46, 50, 53, 63, 105, 106, 108, 109, 111, 113, 115 to 121, 123, 126, 127, 129 to 135, 137 and 140.

[1097] A further aspect ofthe invention provides a method for preparing a compound ofthe formula (II- 3-7) (II-3-7),

[1098]

[1099] characterized in that the method comprises contacting a compound of the formula (l-r)

[1100]

[1101] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-3-7).

[1102] Preferably, the compound of the formula (II-3-7) is a compound of the formula (II-3-7a)

[1103] (II-3-7a).

[1104]

[1105] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 29, 48, 69, 71, 73, 74, 75, 76, 77 and 78. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 29, 48, 69, 71, 73, 74, 75, 76, 77 and 78. 2024P00180WO

[1106] A further aspect of the invention provides a 82. A method for preparing a compound of the formula (II-3-8)

[1107] (II-3-8),

[1108]

[1109] characterized in that the method comprises contacting a compound of the formula (l-s)

[1110]

[1111] with a terpene cyclase enzyme, in particular a meroterpenoid cyclase enzyme, under conditions suitable for the enzyme to produce the compound of the formula (II-3-8).

[1112] Preferably, the compound of the formula (11-3-8) is a compound of the formula (ll-3-8a)

[1113] (ll-3-8a).

[1114]

[1115] In said further aspect of the invention, the terpene cyclase enzyme, in particular the meroterpenoid cyclase enzyme, is as defined above in the first aspect of the invention. In one embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204. In one specific embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In particular, said enzyme has the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a futher embodiment, the terpene cyclase enzyme is a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to SEQ ID NO: 79. In particular, said enzyme has the amino acid sequence of SEQ ID NO: 79.

[1116] Obtention of compounds of the formula (I) in the methods of the invention

[1117] The person skilled in the art is aware that the compounds used as starting material in the method of the invention, i.e. the compounds of the formula (I), are either commercially available or can be synthesized by known methods; either chemically or enzymatically.

[1118] Methods for the enzymatic production leverage metabolic engineering and biocatalysis to produce compounds of the formula (I). Non-limiting Examples of engineered pathways to produce a compound of formula (I), in particular the compounds of formula (l-c), (l-d), (l-e), (l-f), (l-k), (l-m), (l-oa) and (l-ob), are shown in Example 1 and Figure 1.

[1119] Accordingly, a further embodiment of this aspect of the invention is wherein the method further comprises one or more steps prior to step (a), said step(s) comprising preparing a linear terpenoid compound from isopentenyl pyrophosphate (IPP) and dimethyl allyl pyrophosphate (DMAPP) using one or more enzymes having prenyltransferase activity. Said enzymes are well known in the art and nonlimiting examples of such enzymes are disclosed in Example 1.

[1120] Furthermore, the embodiments of Example 1 to produce the compounds of formula (l-c), (l-d), (I-e), (l-f), (l-k), (l-m), (l-oa) and (l-ob) are hereby incorporated by reference.

[1121] In this embodiment of the invention, the precursor compounds to the linear terpenoid compounds are provided from IPP and DMAPP.

[1122] Accordingly, a further embodiment of this aspect of the invention is wherein the method further comprises the steps of preparing IPP and DMAPP as described herein below.

[1123] One means for the preparation of IPP and DMAPP is via the “mevalonate pathway”. The “mevalonate pathway” also known as the “isoprenoid pathway” or “HMG-CoA reductase pathway” is an essential metabolic pathway present in eukaryotes, archaea, and some bacteria. The mevalonate pathway begins with acetyl-CoA and produces two five-carbon building blocks called isopentenyl pyrophosphate (IPP) and dimethyl allyl pyrophosphate (DMAPP). Combining the mevalonate pathway with enzyme activity to generate the terpene precursors GPP, FPP or GGPP allows the recombinant cellular production of terpenes. The pathway is well known in the art. The list of enzymes required for the conversion of acetyl- CoA to IPP and DMAPP is provided below:

[1124] . Acetyl-CoA acetyltransferase (ACAT),

[1125] . 3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase),

[1126] . 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase),

[1127] . Mevalonate kinase,

[1128] . Phosphomevalonate kinase,

[1129] . Mevalonate diphosphate decarboxylase,

[1130] . Isopentenyl diphosphate isomerase.

[1131] An alternative means for the preparation of IPP, and DMAPP is via the methylerythritol phosphate (MEP). The pathway is well known in the art. The list of enzymes required for the conversion of glyceraldehyde 3-phosphate (GAP) and pyruvate to IPP and DMAPP is provided below:

[1132] . 1-Deoxy-D-xylulose 5-phosphate synthase (DXS),

[1133] . 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR),

[1134] . 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (MCT, IspD),

[1135] . 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase (CMK, IspE),

[1136] . 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (MDS, IspF),

[1137] . 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (HDS, IDS),

[1138] . 4-hydroxy-3-methylbut-2-en-1-yl diphosphate reductase (HDR).

[1139] Further alternative pathways to the preparation of IPP and DMAPP are known, see for example: Rinaldi, M. A., et al. (2022). Natural Product Reports 39(1): 90-118. https: / / doi.org / 10.1039 / D1 NP00025J (see part 3 of this article).

[1140] Reaction conditions in the methods of the invention

[1141] The method of the present invention may be an in vivo process or a bioconversion process.

[1142] The term in vivo process (or whole-cell production, or in-vivo production, or in-vivo biosynthesis) refers to a process of using a metabolically active cell where the primary metabolism is active to produce the precursors for the processes of the invention (preferably a microbial cell) to convert a carbon source to a new compound, such as the conversion of a carbon source to a terpene or terpene-derived compound.

[1143] Preferred sources of carbon are sugars, such as mono-, di- or polysaccharides. Very good sources of carbon are for example glucose, fructose, mannose, galactose, ribose, sorbose, ribulose, lactose, maltose, sucrose, raffinose, starch or cellulose. Sugars can also be added to the media via complex compounds, such as molasses, or other by-products from sugar refining. It may also be advantageous to add mixtures of various sources of carbon. Other possible sources of carbon are oils and fats such as soybean oil, sunflower oil, peanut oil and coconut oil, fatty acids such as palmitic acid, stearic acid or linoleic acid, alcohols such as glycerol, methanol or ethanol and organic acids such as acetic acid or lactic acid.

[1144] The cells thus contain all enzymes of one or more biosynthetic pathways. At least some of the enzymes involved in the method are part of the cell's primary metabolism. For example, the cells may contain the enzymes of a pathway to convert a carbon source (e.g., glucose, glycerol, isoprenol, prenol, CO2) to terpenoid precursors (e.g., IPP, DMAPP, FPP) and a pathway converting the terpene precursor to a terpene or terpene derived molecule such as a compound of the formula (I), (A), (B), (C) and / or (D). The enzymes may be present naturally in the cell or the cells can be transformed to produce the enzymes.

[1145] Alternatively, the methods of the present invention may be performed under bioconversion, also known as biotransformation conditions. Bioconversion processes refer to processes of conversion of compounds to different products using a biological process or agent such enzymes or whole cells (preferably a microbial cell). Bioconversion does not include the use of a cell's primary metabolism (as defined above) to produce the precursors for the methods of the invention. A bioconversion process can comprise multistep reactions each performed by a different enzyme. The compounds used in bioconversion process can be extracted from a natural source or produced using a separate chemical or a biochemical process.

[1146] The at least one polypeptide / enzyme which is present during the bioconversion method of the invention or an individual step of the multistep method as defined herein above, can be present in living cells naturally or recombinantly producing the enzyme or enzymes, in harvested cells, dead cells, in permeabilized cells, in crude cell extracts, in purified extracts, or in essentially pure or completely pure form, i.e. under bioconversion conditions. Such extracts may comprise membrane fraction or a liquid fraction prepared from the recombinant host cell that expresses at least one polypeptide / enzyme. The cells may be immobilized on a suitable substrate as is known in the art. At least one polypeptide / enzyme may be present in solution or as an enzyme immobilized on a carrier. One or several enzymes may simultaneously be present in soluble and / or immobilized forms.

[1147] It can be understood by the skilled person that there may be advantages for the use of an in vivo process.

[1148] In particular, a bioconversion process involves multiple steps, typically:

[1149] - Preparation or isolation of the starting compound to be transformed. The compound can be prepared using a chemical or biochemical process or by extraction from a natural source. - Production of the enzymes or (living) cells used for the bioconversion.

[1150] - Biotransformation reaction by contacting the compound with the enzymes or (living) cells. - Product Recovery and Refinement. In comparison, an in vivo process requires a limited number of steps, generally limited to:

[1151] - The cultivation of the microorganism under conditions suitable for the production of the desired compound.

[1152] - Harvesting the cells or growing medium and purification of the desired compound.

[1153] Therefore, the in vivo process is usually more efficient and cost-effective than a bioconversion process.

[1154] Laboratory methods that can be used in in vivo and bioconversion processes of the invention are well known in the art. There follows a discussion on some of the methods that can be used.

[1155] The bioconversion processes according to the invention can be performed in common reactors, which are known to those skilled in the art, and in different ranges of scale, e.g. from a laboratory scale (few milliliters to dozens of liters of reaction volume) to an industrial scale (several liters to thousands of cubic meters of reaction volume). If the polypeptide is used in a form encapsulated by non-living, optionally permeabilized cells, in the form of a more or less purified cell extract or in purified form, a chemical reactor can be used. The chemical reactor usually allows controlling the amount of at least one enzyme, the amount of at least one substrate, the pH, the temperature and the circulation of the reaction medium.

[1156] Where the method of the invention is in vivo then it is preferred that the reaction is performed in a fermenter, where parameters necessary for suitable living conditions for the living cells (e.g. culture medium with nutrients, temperature, aeration, presence or absence of oxygen or other gases, antibiotics, and the like) can be controlled.

[1157] The term "fermentative production" or "fermentation" refers to the ability of a microorganism (assisted by enzyme activity contained in or generated by said microorganism) to produce a chemical compound in cell culture utilizing at least one carbon source added to the incubation.

[1158] The term "fermentation broth" or "fermentation medium" is understood to mean a liquid, particularly aqueous or aqueous / organic solution which is based on a fermentative process and has not been worked up or has been worked up, for example, as described herein.

[1159] Those skilled in the art are familiar with chemical reactors or bioreactors, e.g. with procedures for up-scaling chemical or biotechnological methods from laboratory scale to industrial scale, or for optimizing process parameters, which are also extensively described in the literature (for biotechnological methods see e.g. Crueger und Crueger, Biotechnologie - Lehrbuch der angewandten Mikrobiologie, 2. Ed., R. Oldenbourg Verlag, Munchen, Wien, 1984). The culture medium that is to be used must satisfy the requirements of the particular strains in an appropriate manner. Descriptions of culture media for various microorganisms are given in the handbook " Manual of Methods for General Bacteriology" of the American Society for Bacteriology (Washington D. C., USA, 1981).

[1160] These media that can be used according to the invention may comprise one or more sources of carbon, sources of nitrogen, inorganic salts, vitamins and / or trace elements.

[1161] Sources of nitrogen are usually organic or inorganic nitrogen compounds or materials containing these compounds. Examples of sources of nitrogen include ammonia gas or ammonium salts, such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate or ammonium nitrate, nitrates, urea, amino acids or complex sources of nitrogen, such as corn-steep liquor, soybean flour, soy-bean protein, yeast extract, meat extract and others. The sources of nitrogen can be used separately or as a mixture.

[1162] Inorganic salt compounds that may be present in the media comprise the chloride, phosphate or sulfate salts of calcium, magnesium, sodium, cobalt, molybdenum, potassium, manganese, zinc, copper and iron.

[1163] Inorganic sulfur-containing compounds, for example sulfates, sulfites, di-thionites, tetrathionates, thiosulfates, sulfides, but also organic sulfur compounds, such as mercaptans and thiols, can be used as sources of sulfur.

[1164] Phosphoric acid, potassium dihydrogenphosphate or dipotassium hydrogenphosphate or the corresponding sodium-containing salts can be used as sources of phosphorus.

[1165] Chelating agents can be added to the medium, in order to keep the metal ions in solution. Especially suitable chelating agents comprise dihydroxyphenols, such as catechol or protocatechuate, or organic acids, such as citric acid.

[1166] The fermentation media used according to the invention may also contain other growth factors, such as vitamins or growth promoters, which include for example biotin, riboflavin, thiamine, folic acid, nicotinic acid, pantothenate and pyridoxine. Growth factors and salts often come from complex components of the media, such as yeast extract, molasses, corn-steep liquor and the like. In addition, suitable precursors can be added to the culture medium. The precise composition of the compounds in the medium is strongly dependent on the particular experiment and must be decided individually for each specific case. Information on media optimization can be found in the textbook " Applied Microbiol. Physiology, A Practical Approach" (1997) Growing media can also be obtained from commercial suppliers, such as Standard 1 (Merck) or BHI (Brain heart infusion, DIFCO) etc. All components of the medium are sterilized, either by heating (20 min at 1.5 bar and 121 °C) or by sterile filtration. The components can be sterilized either together or if necessary, separately. All the components of the medium can be present at the start of growing, or optionally can be added continuously or by batch feed.

[1167] The temperature of the culture is normally between 15 °C and 45 °C, preferably 25 °C to 40 °C and can be kept constant or can be varied during the experiment. The pH value of the medium should be in the range from 5 to 8.5, preferably around 7.0. The pH value for growing can be controlled during growing by adding basic compounds such as sodium hydroxide, potassium hydroxide, ammonia or ammonia water or acid compounds such as phosphoric acid or sulfuric acid. Antifoaming agents, e.g. fatty acid polyglycol esters, can be used for controlling foaming. To maintain the stability of plasmids, suitable substances with selective action, e.g. antibiotics, can be added to the medium. Oxygen or oxygencontaining gas mixtures, e.g. the ambient air, are fed into the culture in order to maintain aerobic conditions. The temperature of the culture is normally from 20 °C to 45 °C. Culture is continued until a maximum of the desired product has formed. This is normally achieved within 1 hour to 160 hours.

[1168] Where the method of the invention is a bioconversion, cells containing the at least one enzyme can be permeabilized by physical or mechanical means, such as ultrasound or radiofrequency pulses, French presses, or chemical means, such as hypotonic media, lytic enzymes and detergents present in the medium, or combination of such methods. Examples for detergents are SDS, digitonin, n-dodecylmaltoside, octylglycoside, Triton® X-100, Tween ® 20, deoxycholate, CHAPS (3-[(3-Cholamidopropyl)dimethylammonio]-1-propansulfonate), Nonidet ® P40 (Ethylphenolpoly(ethyleneglycolether), and the like. As stated above, where the method of the invention is an in vivo process, then a detergent is not required for the reasons stated herein.

[1169] The conversion reaction can be carried out batch wise, semi-batch wise or continuously. Reactants (and optionally nutrients) can be supplied at the start of reaction or can be supplied subsequently, either semi-continuously or continuously.

[1170] The bioconversion reaction of the invention, depending on the particular reaction type, may be performed in an aqueous, aqueous-organic or non-aqueous reaction medium.

[1171] An aqueous or aqueous-organic medium may contain a suitable buffer in order to adjust the pH to a value in the range of 5 to 11, like 6 to 10.

[1172] In an aqueous-organic medium an organic solvent miscible, partly miscible or immiscible with water may be applied. Non-limiting examples of suitable organic solvents are listed below. Further examples are mono- or polyhydric, aromatic or aliphatic alcohols, in particular monohydric aliphatic alcohols, most preferably ethanol. The non-aqueous medium may contain is substantially free of water, i.e. will contain less than about 1 wt.-% or 0.5 wt.-% of water.

[1173] Bioconversion methods may also be performed in an organic non-aqueous medium. As suitable organic solvents there may be mentioned aliphatic hydrocarbons having for example 5 to 8 carbon atoms, like pentane, cyclopentane, hexane, cyclohexane, heptane, octane or cyclooctane; aromatic carbohydrates, like benzene, toluene, xylenes, chlorobenzene or dichlorobenzene, aliphatic acyclic and ethers, like diethylether, methyl-tert.-butylether, ethyl-tert. -butylether, dipropylether, diisopropylether, dibutylether; or mixtures thereof.

[1174] The concentration of the reactants / substrates may be adapted to the optimum bioconversion reaction conditions, which may depend on the specific enzyme applied. For example, the initial substrate concentration may be in the 0,1 to 0,5 M, as for example 10 to 100 mM.

[1175] The bioconversion reaction temperature may be adapted to the optimum reaction conditions, which may depend on the specific enzyme applied. For example, the reaction may be performed at a temperature in a range of from 0 to 70 °C, as for example 20 to 50 or 25 to 40 °C. Examples for reaction temperatures are about 30 °C, about 35 °C, about 37 °C, about 40 °C, about 45 °C, about 50 °C, about 55 °C and about 60 °C.

[1176] The bioconversion may proceed until equilibrium between the substrate and then product(s) is achieved, but may be stopped earlier. Usual process times are in the range from 1 minute to 25 hours, in particular 10 min to 6 hours, as for example in the range from 1 hour to 4 hours, in particular 1.5 hours to 3.5 hours. These parameters are non-limiting examples of suitable process conditions.

[1177] In a preferred embodiment of the present invention, the method is performed in a recombinant (host) cell capable of functionally expressing the terpene cyclase enzyme as defined herein above.

[1178] Advantageously, microorganisms such as bacteria, fungi or yeasts are used as host cells. Advantageously, gram-positive or gram-negative bacteria are used, preferably bacteria of the families Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae, Streptomycetaceae, Streptococcaceae or Nocardiaceae, especially preferably bacteria of the genera Escherichia, Pseudomonas, Streptomyces, Lactococcus, Nocardia, Burkholderia, Salmonella, Agrobacterium, Clostridium or Rhodococcus. The genus and species Escherichia coli is quite especially preferred. Furthermore, other advantageous bacteria are to be found in the group of alpha-Proteobacteria, beta-Proteobacteria or gamma-Proteobacteria. Advantageously also yeasts of families like Saccharomyces or Pichia are suitable hosts.

[1179] Preferably, the cell is a bacterium or a fungal cell, in particular a yeast cell. Preferably, the cell is a unicellular organism, a cultured cell derived from a multi-cellular organism, a cell present in a cultured tissue derived from a multicellular organism, or a cell present in a living multicellular organism. Preferably, the cell is a bacterial cell of the genus Escherichia, preferably E. coli, or a yeast cell of the genus Saccharomyces, preferably S. cerevisiae, of the genus Yarrowia, preferably Y. lipolytica, or of the genus Pichia, preferably P. pastoris.

[1180] Alternatively, entire plants or plant cells may serve as natural or recombinant host. As non-limiting examples, the following plants or cells derived therefrom may be mentioned: the genera Nicotiana, in particular Nicotiana benthamiana and Nicotiana tabacum (tobacco); as well as Arabidopsis, in particular Arabidopsis thaliana.

[1181] Product isolation in the methods of the invention

[1182] The methodology of the present invention can further include a step of recovering an end product or an intermediate product, optionally in stereoisomerically or enantiomerically substantially pure form. The term “recovering” includes extracting, harvesting, isolating or purifying the compound from culture or reaction media. Recovering the compound can be performed according to any conventional isolation or purification methodology known in the art including, but not limited to, treatment with a conventional resin (e.g., anion or cation exchange resin, non-ionic adsorption resin, etc.), treatment with a conventional adsorbent (e.g., activated charcoal, silicic acid, silica gel, cellulose, alumina, etc.), alteration of pH, solvent extraction (e.g., with a conventional solvent such as an alcohol, ethyl acetate, hexane and the like), distillation, dialysis, filtration, concentration, crystallization, recrystallization, pH adjustment, lyophilization and the like.

[1183] Identity and purity of the isolated product may be determined by known techniques, like High Performance Liquid Chromatography (HPLC), gas chromatography (GC), Spektroskopy (like IR, UV, NMR), Colouring methods, TLC, NIRS, enzymatic or microbial assays, (see for example: Patek et al. (1994) Appl. Environ. Microbiol. 60:133-140; Malakhova et al. (1996) Biotekhnologiya 11 27-32; und Schmidt et al. (1998) Bioprocess Engineer. 19:67-70. Ullmann's Encyclopedia of Industrial Chemistry (1996) Bd. A27, VCH: Weinheim, S. 89-90, S. 521-540, S. 540-547, S. 559-566, 575-581 und S. 581-587; Michal, G (1999) Biochemical Pathways: An Atlas of Biochemistry and Molecular Biology, John Wiley and Sons; Fallon, A. et al. (1987) Applications of HPLC in Biochemistry in: Laboratory Techniques in Biochemistry and Molecular Biology, Bd. 17.).

[1184] The compounds produced in any of the methods described herein can be converted to derivatives ("derivatives thereof) such as, but not limited to hydrocarbons, esters, amides, glycosides, ethers, epoxides, aldehydes, ketones, alcohols, diols, acetals or ketals. The terpene compound derivatives can be obtained by a chemical method ora biochemical (enzymatic) method. The biochemical conversion can be performed in vivo or in vitro using isolated enzymes, enzymes from lysed cells or bioconversion using whole cells.

[1185] Recombinant cells of the invention

[1186] A further aspect of the invention provides a recombinant cell comprising, capable of producing or producing a compound of the formula (II-1), (II-2), (II-3) and / or (II-4); preferably, a compound of the formula (ll-1a), (ll-1b), (ll-1c), and / or (ll-1d) as defined herewith elsewhere in the description; more preferably, a compound of the formula (11-1-1), (11-1 -1 a), (11-1-2), (II-1-2a), (II-1-3), (11-1 -3a), (11-1-4), (11-1-4a), (11-1-5), (ll-1-5a), (ll-4-6a), (II-Aa), (ll-1b), (ll-1c) and / or (ll-1d).

[1187] Methods for preparing a recombinant cell comprising, capable of producing or producing said compound are provided herein.

[1188] In an embodiment of said aspect of the invention, the cell comprises, is capable of expressing or expresses a terpene cyclase enzyme, preferably a meroterpenoid cyclase enzyme, more preferably a bacterial membrane-integrated meroterpenoid cyclase enzyme. Said meroterpenoid cyclase enzyme preferably comprises at least one or more amino acid motifs selected from:

[1189] [W]xxx[D]xx[ILVMN] (SEQ ID NO: 212),

[1190] PxxAxxxNxxWE (SEQ ID NO: 213),

[1191] MxxxFxxMLxxR (SEQ ID NO: 214),

[1192] RxxxxGQS (SEQ ID NO: 215), and

[1193] NxxMS (SEQ ID NO: 216);

[1194] wherein residues x represent independently of each other any natural amino acid residue.

[1195] In a preferred embodiment of the invention, the cell comprises, is capable of expressing or expresses a meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180. In a further embodiment, the cell comprises a meroterpenoid cyclase enzyme having the amino acid sequence of any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180.

[1196] In a further embodiment of the invention, the cell comprises, is capable of expressing or expresses a mutant bacterial membrane-integrated meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180; wherein the mutant meroterpenoid cyclase enzyme has an amino acid substitution at amino acid position 9 relative to the sequence provided in SEQ ID NO: 29.

[1197] With reference to the above embodiments, the substitution at amino acid position 9 relative to the sequence provided in SEQ ID NO: 29 may be a substitution to cysteine, methionine or threonine.

[1198] In an alternative or yet a further embodiment of the invention, the cell comprises, is capable of expressing or expresses a mutant bacterial membrane-integrated meroterpenoid cyclase enzyme having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 204; preferably, to any one of SEQ ID NOs: 1 to 141 and 162 to 180; wherein the mutant meroterpenoid cyclase enzyme has an amino acid substitution at amino acid position 123, 126 and / or 165 relative to the sequence provided in SEQ ID NO: 74. With reference to the above embodiments, the substitution at amino acid position 123, 126 and / or 165 relative to the sequence provided in SEQ ID NO: 74 may be a substitution to methionine, serine or glycine.

[1199] The recombinant cell may be any such cell suitable for the production of a compound of the formula (II-1), (II-2), (II-3) and / or (II-4).

[1200] A list of suitable cells for the production of a compound of the formula (II-1), (II-2), (II-3) and / or (II-4) is provided above in relation to the method of the invention and are also cells for this aspect of the invention.

[1201] Preferably, the cell is a bacterium or a fungal cell, in particular a yeast. Preferably, the cell is a unicellular organism, a cultured cell derived from a multi-cellular organism, a cell present in a cultured tissue derived from a multicellular organism, or a cell present in a living multicellular organism. Preferably, the cell is a bacterial cell of the genus Escherichia, preferably E. coli, of the genus Pseudomonas, preferably Pseudomonas alloputida, or a yeast cell of the genus Saccharomyces, preferably S. cerevisiae, of the genus Yarrowia, preferably Y. lipolytica, or of the genus Pichia, preferably P. pastoris.

[1202] Methods of introducing recombinant nucleic acid sequences into such host cells are well known in the art and constitute routine laboratory methodologies which do not need to be further described herein.

[1203] An embodiment of this aspect of the invention is wherein the cell of the invention further comprises, is capable of expressing or expresses one or more prenyltransferase enzyme(s).

[1204] A further embodiment of this aspect of the invention is wherein the cell of the invention comprises, is capable of expressing or expresses enzymes for providing IPP and DMAPP. Said compound may be provided via the “mevalonate pathway”, methylerythritol phosphate (MEP) pathway or alternative pathways for the preparation of IPP and DMAPP.

[1205] In one embodiment of the invention, the cell comprises, is capable of expressing or expresses enzymes of the mevalonate pathway:

[1206] . Acetyl-CoA acetyltransferase (ACAT)

[1207] . 3-hydroxy-3-methylglutaryl-CoA synthase (HMG-CoA synthase)

[1208] . 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA reductase)

[1209] . Mevalonate kinase

[1210] . Phosphomevalonate kinase

[1211] . Mevalonate diphosphate decarboxylase

[1212] . Isopentenyl diphosphate isomerase

[1213] . Dimethylallyl diphosphate synthase In a further embodiment of the invention, the cell comprises, is capable of expressing or expresses enzymes of the MEP pathway:

[1214] . 1-Deoxy-D-xylulose 5-phosphate synthase (DXS)

[1215] . 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR)

[1216] . 2-C-methyl-D-erythritol 4-phosphate cytidylyltransferase (MCT, IspD)

[1217] . 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase (CMK, IspE)

[1218] . 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase (MDS, IspF)

[1219] . 4-hydroxy-3-methylbut-2-en-1-yl diphosphate synthase (HDS, IDS)

[1220] . 4-hydroxy-3-methylbut-2-en-1-yl diphosphate reductase (HDR)

[1221] Cell culture fermentation media of the invention

[1222] A further aspect of the invention provides a cell culture fermentation medium comprising a compound of the formula (II-1), (II-2), (II-3) and / or (II-4); preferably, a compound of the formula (11-1 a), (II-1b), (11-1 c), and / or (11-1 d) as defined herewith elsewhere in the description; more preferably, a compound of the formula (11-1-1), (11-1-1 a), (11-1-2), (II-1-2a), (II-1-3), (ll-1-3a), (11-1-4), (II-1-4a), (11-1-5), (11-1 -5a), (II-4-6a), (II-Aa), (ll-1b), (ll-1c) and / or (ll-1d).

[1223] Accordingly, a further aspect of the invention provides a cell culture fermentation medium comprising the recombinant cell of the invention as described herein above.

[1224] The cell culture fermentation media can be a nutrient rich broth for the growth and maintenance of the cells during the production phase. Yeast culture conditions for maintaining and propagating various strains can require specific formulations of complex media for use in cloning and protein expression, and can be appreciated by those of skill in the art. Commercially available culture media can be used from ThermoFisher for example. The media can be YPD broth or can have a yeast nitrogen base. Yeast can be grown in YPD or synthetic media at 30 °C.

[1225] Lysogeny broth (LB) is typically used for bacterial cells. The bacterial cells can have antibiotic resistance to prevent the growth of other cells in the culture media and contamination.

[1226] Reaction mixtures of the invention

[1227] The method of the invention may be a bioconversion process.

[1228] Accordingly, an aspect of the present invention is a reaction mixture comprising a compound of the formula (11-1), (II-2), (II-3) and / or (II-4); preferably, a compound of the formula (II-1a), (II-1b), (II-1c), and / or (II-1d) as defined herewith elsewhere in the description; more preferably, a compound of the formula (II-1-1), (II-1-1a), (II-1-2), (II-1-2a), (II-1-3), (II-1-3a), (II-1-4), (II-1-4a), (II-1-5), (II-1-5a), (II-4-6a), (II-Aa), (II-1b), (II-1c) and / or (II-1d).

[1229] Additional components of the reaction mixture may include detergents, co-factors, cells, cell-debris, cell culture media, and other such components well known to the person skilled in the art.

[1230] New compounds of the invention

[1231] A further aspect of the invention provides a compound obtained or obtainable by a method of the invention or from a recombinant cell of the invention or from a cell culture fermentation medium of the invention or from a reaction mixture of the invention as described herein above.

[1232] Another futher aspect of the invention provides a compound selected from the group consisting of the compounds of the formula (11-1 a), (11-1 b), (11-1 c) and (ll-1d); preferably, a compound selected from the group consisting of the compounds of the formula

[1233]

[1234] (ll-1-5a)

[1235] (11-1 c), and

[1236]

[1237] (Il-1d), or a derivative thereof;

[1238] wherein R2= H or an alcohol protecting group, particularly

[1239]

[1240] ' R3, preferably H;

[1241] R3represents a C1-4alkyl group, preferably CH3; and

[1242] wherein any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration; and

[1243] the single dotted line represents the bond by which said substituent is bound to the rest of a molecule.

[1244] Also preferably, a further aspect of the invention provides a cyclized compound prepared by a cyclisation of a compound of the formula (I) in the presence of a meroterpenoid cyclase enzyme n = 0 or 1 or 2; R represents

[1245]

[1246] wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;

[1247] wherein R2= H or an alcohol protecting group, particularly

[1248]

[1249] preferably H, R3represents a C1-4alkyl group, preferably CH3; and

[1250] m = 0 or 1 or 2;

[1251] z = 1 or 2;

[1252] R° independently from each other represents H or CH3or CH2OH;

[1253] wherein

[1254] any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;

[1255] and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond;

[1256] with the proviso that the cyclized compound is not

[1257]

[1258] Use of compounds of the invention

[1259] A further aspect of the invention comprises the use of a compound according to any of the previous embodiments of the invention as a perfumery, flavor or aroma ingredient, or as a precursor thereof.

[1260] As mentioned above, the invention comprises the use of a compound of the invention as a perfuming ingredient, or as a precursor thereof. In other words, it concerns a method or a process to confer, enhance, improve or modify the odor properties of a perfuming composition or of a perfumed article or of a surface, which method comprises adding to said composition or article an effective amount of at least a compound of the invention, e.g. to impart its typical note. Understood that the final hedonic effect may depend on the precise dosage and on the organoleptic properties of the invention’s compound, but anyway the addition of the invention’s compound will impart to the final product its typical touch in the form of a note, touch or aspect depending on the dosage.

[1261] By “use of a compound ...

Claims

1. 2024P00180WO CLAIMS1. A method for preparing a compound of the formula (II-1), (II-2), (II-3) and / or (II-4),3.

4. and / or5.in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises more than one--OH group,6.a compound having at least one--O-- group,7.which is formally obtained by an intramolecular etherification or esterification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification or esterification, two--OH groups form, under cleavage of a molecule of water, a cyclic--O-- group; and / or8.in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises at least two--OH groups and at least one carbonyl group,9.a compound having at least one--O-- group,10.which is formally obtained by ketalization of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the ketalization a carbonyl group and two--OH groups form, under cleavage of water, a cyclic--O-- group;11.and / or12.in the case where the compound of the formula (II-1) or (II-2) or (II-3) or (II-4) comprises at least one--OH group and at least one carbonyl group,13.a compound having at least one--O-- group,14.which is formally obtained by etherification of compound of the formula (II-1) or (II-2) or (II-3) or (II-4), wherein in the etherification an--OH group a carbonyl group having undergone enolization form, under cleavage of water, a cyclic--O-- group; or a derivative thereof,15.with the proviso that said compound is not17. 19.wherein20.A represents either21. 23.A' represents H, or24.

25. wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;26.O27.wherein R2= H or an alcohol protecting group, particularly28.

29. '' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and30.m = 0 or 1 or 2;31.n = 0 or 1 or 2;32.with the proviso that in formula (II-3) and / or (II-4) n is 1 or 2;33.z = 1 or 2;34.R° independently form each other represents H or CH3or CH2OH;35.wherein36.any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;37.and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond;38.characterized in that the method comprises:39.(a) contacting a compound of the formula (I)41.

42. with a meroterpenoid cyclase enzyme under conditions suitable for the meroterpenoid cyclase enzyme to produce the compound of the formula (II-1), (II-2), (II-3) and / or (II-4).

2. The method according to claim 1, characterized in that the cyclic--O-- group is part of a 4-6 membered ring.

3. The method according to any of claims 1 and 2, characterized in that the cyclic--O-- group is part of a cyclic ether or a cyclic ester (lactone).

4. The method according to any of the preceding claims, characterized in that the compound of having at least one--O-- group is selected from the group consisting of the compounds of the formula46.(ll-B),47.(Il-D),49.

50. The method according to any of the preceding claims, characterized in that the compound of the formula (I) is selected from the group consisting of the group consisting of the compounds of the formula52.

54.

6. The method according to any of the preceding claims, characterized in that the meroterpenoid cyclase enzyme is a bacterial membrane-integrated meroterpenoid cyclase enzyme, preferably comprising at least one or more amino acid motifs selected from:[W]xxx[D]xx[ILVMN] (SEQ ID NO: 212),57.PxxAxxxNxxWE (SEQ ID NO: 213),58.MxxxFxxMLxxR (SEQ ID NO: 214),59.RxxxxGQS (SEQ ID NO: 215), and60.NxxMS (SEQ ID NO: 216);61.wherein residues x represent independently of each other any natural amino acid residue.

7. The method according to any of the preceding claims, characterized in that the meroterpenoid cyclase enzyme has at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity to any one of SEQ ID NOs: 1 to 141 and 162 to 180.

8. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-a) and the compound of the formula (II-2) is a compound of the formula (11-2-17)64.(l-a),65.(II-2-17);67. 69.preferably, a compound of the formula (11-2-17a)70.(11-2-17a).

72.

9. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-b) and the compound of the formula (11-1), (II-2), (II-3) and / or (II- 4) is a compound of the formula (II-1-3), (II-2-7), (II-2-8), (II-2-9), (II-3-2) and / or (II-4-3)75.(l-b),76.

77. (II-1-3),78.(II-2-7),79.(II-2-8),80.(II-2-9),81.(II-3-2),82.(II-4-3);84.

85. preferably, a compound of the formula (11-1 -3a), (I l-2-7a), (ll-2-8a), (II-2-9), (ll-3-2a) and / or (I l-4-3a)86.(ll-1-3a),87.(ll-2-7a),88.

89. (ll-2-8a),90.(II-2-9),91.(II-3-2a),92.(II-4-3a).

94.

95.

10. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-c) and the compound of the formula (II-2), (II-3) and / or (II-4) is a compound of the formula (11-2-10), (II-3-3) and / or (II-4-4)97.

98. (II-4-4);100. 102.preferably, a compound of the formula (II-2-10a), (II-3-3a) and / or (II-4-4a)103.(II-2-10a),104.(ll-3-3a),105.(ll-4-4a).

107.

108.

11. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-d) and the compound of the formula (11-1), (II-2), (II-3) and / or (II- 4) is a compound of the formula (11-1-2), (II-2-4), (II-2-5), (II-A), (11-4-1) and / or (II-4-2)109.(l-d),110.(II-1-2),112.

113. (II-2-4),114.(II-2-5),115.(II-A),116.(II-4-1),117.(II-4-2);119.

120. preferably, a compound of the formula (ll-1-2a), (II-2-4a), (II-2-5), (II-Aa), (11-4-1 b) and / or (ll-4-2b)121.(ll-1-2a),122.

123. (II-2-4a),124.(II-2-5),125.(II-Aa),126.(II-4-1b),127.(II-4-2b).

129.

12. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-e) and the compound of the formula (11-1), (II-2), (II-3) and / or (II- 4) is a compound of the formula (11-1-1), (11-2-1), (II-2-2), (II-2-3), (11-3-1) (11-4-1) and / or (II-4-2)'OH (l-e),132.(II-1-1),133.(II-2-1),134.(II-2-2),135.(II-2-3),136.(II-3-1),137.(II-4-1),138.

139. (II-4-2);141. 143.preferably, a compound of the formula (II-1-1a), (II-2-1), (II-2-2a), (II-2-3a), (II-3-1a) (II-4-1a) and / or (II-4-2a)144.(II-1-1a),145.(II-2-1),146.(ll-2-2a),147.OH (ll-2-3a),148.(11-3-1 a),150.

151. (11-4-1 a),152.(ll-4-2a).

154.

13. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-f) and the compound of the formula (II-2) is a compound of the formula (II-2-4)157.(l-f),158.(II-2-4);160.

161. preferably, a compound of the formula (ll-2-4b)162.(ll-2-4b).

164.

165.

14. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-g) and the compound of the formula (11-1) is a compound of the formula (ll-EB) o (I-g),166.(II-B);168. 170.preferably, a compound of the formula (II-Ba)171.(II-Ba).

173.

15. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-h) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (I l-C) and / or (II-4-5)177. 179.preferably, a compound of the formula (ll-Ca) and / or (II-4-5a) (II-Ca),180.(ll-4-5a).

182.

16. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-i) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (II-C), (II-4-5) and / or (II-4-6)185.(l-i),186.(II-C),187.(II-4-5),189.

190. (II-4-6);192. 194.preferably, a compound of the formula (II-Cb), (II-4-5b) and / or (ll-4-6a)195.(II-Cb),196.(II-4-5b),197.(II-4-6a).

199.

17. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-j) and the compound of the formula (II-3) is a compound of the formula (ll-D)202.(l-j),203.

204. 2024P00180WO205.(II-D);207. 209.preferably, a compound of the formula (II-Da)210.(II-Da).

212.

18. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-k) and the compound of the formula (11-1) and / or (II-2) is a compound of the formula (11-1-4), (11-2-12), (11-2-13) and / or (11-2-14)215.(II-1-4),216.(11-2-12),217.(11-2-13),219.

220. (11-2-14);222. 224.preferably, a compound of the formula (11-1 -4a), (11-2-12a), (11-2-13) and / or (11-2-14a)225.(II-1-4a),226.(II-2-12a),227.(11-2-13),228.(II-2-14a).

230.

231.

19. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-l) and the compound of the formula (II-3) is a compound of the formula (ll-E)233.

234. o (II-E);236. 238.preferably, a compound of the formula (II-Ea)239.(II-Ea).

241.

20. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-m) and the compound of the formula (11-1) and / or (II-2) is a compound of the formula (11-1-5), (11-2-15) and / or (11-2-16)244.(II-1-5),245.(II-2-15),246.(11-2-16);248.

249. preferably, a compound of the formula (II-1-5a), (II-2-15a) and / or (II-2-16) (II-1-5a),250.(11-2-15a),251.(11-2-16).

253.

254.

21. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-n) and the compound of the formula (II-3) is a compound of the formula (II-3-5)255.(l-n),256.(II-3-5);258.

259. preferably, a compound of the formula (II-3-5a)260.(II-3-5a).

262.

22. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-o) and the compound of the formula (11-1) is a compound of the formula (ll-F)(l-o),265.(II-F);267. 269.preferably, the compound of the formula (I) is a compound of the formula (l-oa) and the compound of the formula (II-F) is a compound of the formula (II-Fa)270.(l-oa),271.(II-Fa);273.

274. also preferably, the compound of the formula (I) is a compound of the formula (l-ob) and the compound of the formula (II-F) is a compound of the formula (II-Fb)275.(l-ob),276.(II-Fb).

278.

279.

23. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-p) and the compound of the formula (11-2) and / or (11-3) is a compound of the formula (11-2-11) and / or (II-3-4)280.(I-p),281.

282. (11-2-11),283.(II-3-4);285. 287.preferably, a compound of the formula (11-2-11a) and / or (ll-3-4a)288.OH289.(II-2-11a),290.(II-3-4a).

292.

293.

24. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-q) and the compound of the formula (II-3) and / or (II-4) is a compound of the formula (II-3-6) and / or (II-4-7)295.

296. (II-3-6),297.(II-4-7);299. 301.preferably, a compound of the formula (ll-3-6a) and / or (II-4-7a)302.(II-3-6a),303.(II-4-7a).

305.

25. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-r) and the compound of the formula (II-3) is a compound of the formula (II-3-7)309.

310. (II-3-7);312. 314.preferably, a compound of the formula (II-3-7a)315.(II-3-7a).

317.

26. The method according to any of claims 1 to 7, characterized in that the compound of the formula (I) is a compound of the formula (l-s) and the compound of the formula (II-3) is a compound of the formula (II-3-8)320.(II-3-8);322. 324.preferably, a compound of the formula (ll-3-8a) (ll-3-8a).

326.

27. The method according to any of the preceding claims, characterized in that the process is an in vivo or a bioconversion process.

28. The method according to any of the preceding claims, characterized in that the process is performed in a recombinant cell capable of functionally expressing the meroterpenoid cyclase enzyme; preferably, said recombinant cell is a bacterial cell, a plant cell, a fungal cell such as a yeast cell; more preferably, said recombinant cell is of the genus Escherichia, Pseudomonas, Saccharomyces, Yarrowia or Pichia.

29. A compound of the formula (11-1), (II-2), (II-3) and / or (II-4) obtained or obtainable by the method according to any of claims 1 to 28.

30. A compound selected from the group consisting of332.(II-1-1), (ll-1-1a),334.

335. (II-1-2), (II-1-2a) (11-1-3) (11-1-4) (ll-1-4a336.

337. (H-1 b), (11-1 c), and 2024P00180WO339. 341.(ll-1d),342.or a derivative thereof;343.wherein R2= H or an alcohol protecting group, particularly344.

345. '' R3, preferably H;346.R3represents a C1-4alkyl group, preferably CH3; and347.wherein any wavy line represents a carbon-carbon bond which is either in the Z- or in the E- configuration, preferably in the E-configuration; and348.the single dotted line represents the bond by which said substituent is bound to the rest of a molecule.

31. Cyclized compound prepared by a cyclisation of a compound of the formula (I) in the presence of a meroterpenoid cyclase enzyme350.A represents either or n = 0 or 1 or 2;352.

353. O354.wherein R2= H or an alcohol protecting group, particularly355.

356. '' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and357.m = 0 or 1 or 2;358.z = 1 or 2;359.R° independently form each other represents H or CH3or CH2OH;360.wherein361.any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;362.and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond; with the proviso that the cyclized compound is not364.

32. A recombinant cell comprising a compound according to any of claims 29 to 31.

33. Use of a compound according to any of claims 29 to 31 as a perfumery, flavor or aroma ingredient, or as a precursor thereof.

34. Use of a meroterpenoid cyclase enzyme for the preparation of a compound according to any of claims 29 to 31.

35. Use of a meroterpenoid cyclase enzyme for the cyclisation of a compound of the formula (I)371.

372. wherein373.A represents either374. 376.n = 0 or 1 or 2;377.R represents379.

380. wherein R1= H, OH, a C1-8-alkyl group or a C1-12-alkoxy group;381.wherein R2= H or an alcohol protecting group, particularly382.

383. '' R3, preferably H, R3represents a C1-4alkyl group, preferably CH3; and384.m = 0 or 1 or 2;385.z = 1 or 2;386.R° independently form each other represents H or CH3or CH2OH;387.wherein388.any dotted line represents the bond by which the substituent is bound to the rest of the molecule any wavy line represents a carbon-carbon bond which is either in the Z- or in the E-configuration, preferably in the E-configuration;389.and wherein any double bond having dotted line ( - ) represents independently from each other either a single carbon-carbon bond or a carbon-carbon double bond; with the proviso that the compound does not comprise two cumulated carbon-carbon double bonds and that at least one of the double bond having dotted line ( - ) represents a carbon-carbon double bond; with the proviso that the compound of the formula (I) is not391.

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