Polypeptide for catalyzing cyclization of lignans, coding gene, preparation method and application

Abstract

The invention discloses a polypeptide for catalyzing cyclization of lignans, a coding gene, a preparation method and application, and belongs to the technical field of gene engineering and enzyme engineering. The polypeptide for catalyzing cyclization of lignans is polypeptide SchCYP719G1b, and the amino acid sequence of the polypeptide is selected from (1) an amino acid sequence as shown in SEQ ID No.1; (2) an amino acid sequence which is generated by substituting, deleting and / or adding one or more amino acids to the amino acid sequence as shown in SEQ ID No.1 and has the same catalytic activity as (1); or (3) a truncated body of (1) or (2) having the same catalytic activity as (1). The polypeptide SchCYP719G1b can catalyze C-C coupling on a benzene ring of dibenzyl butane type lignan, and lays an important foundation for biosynthesis and drug development of biphenyl cyclooctene type lignan.

Description

Technical Field

[0001] The invention belongs to the technical field of genetic engineering and enzyme engineering, and specifically relates to a polypeptide catalyzing the cyclization of lignans and a coding gene, a preparation method and an application. Background Art

[0002] Schisandra chinensis ( Schisandra chinensis ) is a woody plant of the genus Schisandra in the family Schisandraceae. Schisandra has a wide range of medicinal values, including liver protection, anti-tumor, anti-viral, antioxidant, neuroprotective and other effects. Biphenylcyclooctene lignans are one of the important categories of lignan compounds. They are characterized by containing a highly oxidized biphenylcyclooctene (eight-membered ring) structure. This type of lignan is concentrated in the family Schisandraceae and is also the active ingredient of Schisandraceae medicinal plants.

[0003] In recent years, studies have shown that biphenylcyclooctene lignans have rich and significant pharmacological activities and are potential sources of drugs, mainly showing liver protection, antiviral and neuroprotective effects. For example, schisandrin B has entered Phase II clinical trials as a natural biphenylcyclooctene lignan for the treatment of drug-induced cardiotoxic damage; (−)-gomisin M1 can inhibit the proliferation, migration and invasion of liver cancer cells by regulating the biogenesis of miRNA; (+)-gomisin M2, gomisin J and schisandrin A have been shown to inhibit the activity of breast cancer cells.

[0004] Biphenylcyclooctene lignans have a special structure, and their natural eight-membered ring structure is difficult to chemically synthesize. Therefore, the current acquisition of this type of compound mainly relies on plant extraction and separation, which seriously restricts the development of drugs. The study of the biosynthesis of biphenylcyclooctene lignans can greatly promote the exploration and utilization of medicinal molecules in this type of lignans. The key step is the formation of the eight-membered ring mother nucleus. The analysis of this step lays an important foundation for the subsequent biosynthesis of various biphenylcyclooctene lignans and drug development. Summary of the invention

[0005] In view of the above-mentioned prior art, the present invention provides a polypeptide and encoding gene for catalyzing the cyclization of lignans, a preparation method and application, and provides a new polypeptide that can catalyze the cyclization of lignans, which can catalyze the CC coupling on the benzyl ring of dibenzylbutane-type lignans, laying an important foundation for the biosynthesis of biphenylcyclooctene-type lignans and drug development.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is: to provide a polypeptide that catalyzes the cyclization of lignans, the polypeptide that catalyzes the cyclization of lignans is a polypeptide Sch CYP719G1b, whose amino acid sequence is selected from: (1) the amino acid sequence shown in SEQ ID No. 1; (2) an amino acid sequence produced by substitution, deletion and / or addition of one or more amino acids to the amino acid sequence shown in SEQ ID No. 1, and having the same catalytic activity as (1); or (3) A truncated form of (1) or (2) having the same catalytic activity as (1).

[0007] The present invention also discloses a gene encoding a polypeptide that catalyzes the cyclization of lignans. Sch The coding gene sequence of CYP719G1b is selected from: (1) the nucleotide sequence shown in SEQ ID NO.2; (2) a nucleotide sequence having more than 80% homology with the nucleotide sequence shown in SEQ ID NO.2, and the encoded polypeptide has the same catalytic activity as the polypeptide encoded by (1); or (3) A truncated form of (1) or (2) having the same catalytic activity as the polypeptide encoded by (1).

[0008] Furthermore, an expression vector containing the above encoding gene.

[0009] Furthermore, a recombinant microorganism or a transgenic cell line containing the above encoding gene.

[0010] Furthermore, the preparation method of the polypeptide that catalyzes the cyclization of lignans comprises the following steps: introducing the above-mentioned coding gene into a receptor cell, expressing and extracting the polypeptide, and obtaining the polypeptide.

[0011] Furthermore, the use of the above-mentioned polypeptide catalyzing the cyclization of lignans is to Sch CYP719G1b is used to catalyze the cyclization of dibenzylbutane-type lignans.

[0012] Furthermore, peptide Sch CYP719G1b catalyzes the CC coupling in biphenylcyclooctene-type lignans.

[0013] Furthermore, peptide Sch CYP719G1b catalyzes the conversion of progomisin to gomisin J.

[0014] The beneficial effect of the present invention is that: the present invention provides a polypeptide belonging to the CYP450 family Sch CYP719G1b and its encoding gene, polypeptide Sch CYP719G1b can participate in the formation of the eight-membered ring skeleton of Schisandra biphenylcyclooctene lignans, and can catalyze the formation of biphenylcyclooctene lignan gomisin J through the CC coupling of the benzylbutane lignan. SchCYP719G1b can convert dibenzylbutane lignans into biphenylcyclooctene lignans, which can be used for the skeleton biosynthesis of biphenylcyclooctene lignans, laying a foundation for the synthetic biology research of various biphenylcyclooctene lignans in Schisandra chinensis, and has great application value. Sch The compound obtained by CYP719G1b is an active ingredient in medicine, or the compound obtained is used as an intermediate for subsequent synthesis reaction to obtain an active ingredient in medicine, a peptide Sch CYP719G1b has positive application prospects in the pharmaceutical industry. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1 To express peptides Sch Recombinant vector map of CYP719G1b; Figure 2 For peptide Sch LC-MS spectrum of the enzyme activity catalyzed reaction of CYP719G1b using pregomisin as substrate; Figure 3 For peptide Sch CYP719G1b catalyzes the reaction process of converting pro-gomisin to gomisin J; Figure 4 For peptide Sch Relative activities of CYP719G1b mutants compared to the wild type. DETAILED DESCRIPTION

[0016] The specific implementation modes of the present invention are described in detail below with reference to the embodiments.

[0017] Example 1 The steps of transcriptome-based screening of Schisandra CYP450 genes are as follows: (1) UPLC was used to detect the differences in the content of dibenzylbutane-type lignans in Schisandra chinensis fruit, mature stems, old leaves and roots.

[0018] (2) Based on the genome of Schisandra chinensis, the members of the Schisandra chinensis CYP450 gene family were identified, and the HMM model of the CYP450 gene family was used to preliminarily screen the key enzyme protein sequences of CYP450 in Schisandra chinensis.

[0019] (3) Based on the transcriptome data of different tissues of Schisandra chinensis, differential gene expression was analyzed. Correlation analysis was performed on the metabolome and transcriptome. By calculating the Pearson correlation coefficient, genes with high correlation coefficients with gomisin J were screened as candidate genes to obtain expressed peptides. Sch CYP719G1b gene; polypeptide Sch The amino acid sequence of CYP719G1b is shown in SEQ ID No. 1, encoding a polypeptideSch The gene sequence of CYP719G1b is shown in SEQ ID No.2.

[0020] Example 2 Expressed peptide Sch Cloning of CYP719G1b gene 1. Extraction of total RNA from Schisandra chinensis by CTAB-PVP method (1) Take fresh Schisandra chinensis plant material (leaves or fruits) and quickly grind them into powder in liquid nitrogen.

[0021] (2) Use a centrifuge tube to estimate the amount of 100 mg of powder into a pre-cooled 2 mL centrifuge tube, add 800 μL of 65°C preheated CTAB-PVP extraction buffer, and place it on a vortex oscillator for 30 seconds to fully lyse it; The CTAB-PVP extraction buffer contains 100 mM Tris·HCl (pH 8.0), 2% CTAB (w / v), 2% PVP (polyvinyl pyrrolidone, w / v), 25 mM EDTA, 2 M NaCl, and mercaptoethanol is added to a final concentration of 0.2% (v / v) after high-pressure steam sterilization; the solution is prepared with double distilled water (ddH2O) treated with DEPC and sterilized by high pressure.

[0022] (3) Place in a 65°C water bath for 30 min, inverting and mixing every 10 min.

[0023] (4) After cooling to room temperature, add 800 μL of chloroform, invert to mix, and centrifuge at 13,000 rpm for 10 min at 4°C.

[0024] (5) Transfer the supernatant obtained by centrifugation to a new 2 mL centrifuge tube, add 800 μL of chloroform, shake to mix evenly, and centrifuge at 13,000 rpm for 10 min at 4 °C.

[0025] (6) Repeat step (5) once.

[0026] (7) Carefully pipette the supernatant obtained by centrifugation into a new 1.5 mL centrifuge tube, add 8 M LiCl solution (1 / 3 of the supernatant volume), and let stand at -20 °C overnight.

[0027] (8) Centrifuge at 13,000 rpm for 10 min at 4°C and discard the supernatant.

[0028] (9) Add 700 μL of 75% (v / v) ethanol (prepared with DEPC water) to wash the precipitate, and then centrifuge at 13,000 rpm for 10 min at 4°C; repeat the washing of the precipitate three times. After the third centrifugation, discard the supernatant and evaporate the remaining ethanol.

[0029] (10) Add 30 μL of sterile water treated with Proteinase K to dissolve the RNA and obtain total RNA. Use a BioPhotometer plus nucleic acid protein analyzer to measure the concentration and quality of the extracted RNA.

[0030] 2. Encoding polypeptide Sch Full-length amplification of the CYP719G1b gene (1) Primer design Using the software SnapGene to encode polypeptides Sch Full-length primers SchCYP719G1b-F / R were designed on both sides of the open reading frame (ORF) of the CYP719G1b gene to amplify the gene; The sequence of SchCYP719G1b-F is: CTATAGGGCCCGGGATGGAGGGCCAATGGGTTGT (SEQ ID No. 3); The sequence of SchCYP719G1b-R is: CTAGACTTCAGGTTGTCTAACTCCTTTCCTTTTCGG (SEQ ID No. 4).

[0031] (2) cDNA synthesis The total RNA of Schisandra chinensis obtained in the above step was used as a template, and the cDNA template chain was obtained by PCR technology using the PrimerScript RT Master Mix reverse transcription system. The reverse transcription system is shown in Table 1, and the reverse transcription program is: 37°C, 15min; 85°C, 15s. The reverse transcription product was stored at -20°C and diluted before use.

[0032] Table 1 Reverse transcription PCR system

[0033] (3) Target gene amplification The Schisandra cDNA obtained in the above step was diluted and used as a template, and SchCYP719G1b-F / R was used as a primer for amplification. The amplification system is shown in Table 2, and the amplification program is shown in Table 3. The PCR reaction product was detected by agarose gel electrophoresis and the band of the target size was cut and recovered to obtain the expressed polypeptide. Sch Gene sequence of CYP719G1b.

[0034] Table 2 Target gene amplification system

[0035] Table 3 Target gene amplification procedure

[0036] Example 3 Peptides Sch Construction of expression vector of CYP719G1b 1. Amplify the expressed peptide with homology arms Sch CYP719G1b gene fragment The peptide was expressed using the primer pair SchCYP719G1b-pESC-His-F / R with homology arms Sch The gene fragment of CYP719G1b is amplified, and the amplification system and procedure are as shown in Table 2 and Table 3 in Example 1. The PCR reaction product is detected by agarose gel electrophoresis and the band of the target size is cut and recovered to obtain the target fragment with homology arms; The sequence of SchCYP719G1b-pESC-His-F is: CCGTAATACGACTCACTATAGGGCCCGGGATGGAGGGCCAATGGGTTGT (SEQ ID No. 5); The sequence of SchCYP719G1b-pESC-His-R is: TAAATAGGGACCTAGACTTCAGGTTGTCTAACTCCTTCCTTTTCGG (SEQ ID No. 6).

[0037] 2. Construction of homologous recombination expression vector (1) Double digestion of vector: vector pESC-His Sal I and Xho I was used for enzyme digestion. The double enzyme digestion system is shown in Table 4. After the double enzyme digestion system was reacted in a 37°C water bath for 30 min, 10× loading buffer was added to the digestion product to terminate the reaction, and then agarose gel electrophoresis was performed and the band of the correct size was selected for gel recovery to obtain the double enzyme digestion vector.

[0038] Table 4 Double enzyme digestion system

[0039] (2) Homologous recombination, transformation and positive verification: The target fragment with homology arms was connected to the double-enzyme-cut vector pESC-His using a homologous recombination kit (ClonExpress Ultra One Step Cloning Kit V2). The system is shown in Table 5.

[0040] Table 5 Homologous recombination ligation system

[0041] After the above components are fully mixed, react at 37°C for 30 minutes, and immediately cool on ice; transform the ligation product into E. coli DH5α competent cells. The specific steps are: take out the E. coli DH5α competent cells stored at -80°C and thaw them on ice, add all the ligation products, gently blow and mix, and place on ice for 30 minutes; heat shock in a 42°C water bath for 45 seconds and quickly place on ice for 2 minutes, add 500μL of antibiotic-free LB medium and shake and culture in a 37°C incubator for 1 hour, take 200μL of the transformation solution and spread it on LB solid medium (containing 100μg / mL ampicillin), and culture it in a 37°C incubator for 12h~16h.

[0042] Single clones were selected for colony PCR verification. A bright and single target size band was amplified as a positive single clone. The positive clone was sent for sequencing to encode a polypeptide. Sch The gene sequence of CYP719G1b is shown in SEQ ID No. 2. The correct sequenced monoclonal stock was taken and the SchCYP719G1b-pESC-His plasmid was extracted; the plasmid map is shown in Figure 1 As shown, a total of 8179 bp, ori in the figure indicates the starting point of plasmid replication, Figure 1 There are ori, 2µ ori and f1 ori in the sequence; HIS3 represents the histidine synthesis gene; SchCYP719G1b is the inserted coding polypeptide Sch The gene sequence of CYP719G1b; AmpR is the ampicillin resistance gene; promoter indicates the promoter, Figure 1 There are HIS3 promoter, GAL,10 promoter, AmpR promoter; terminator means terminator, there is ADH1 terminator in the figure; MCS is the multiple cloning site, Figure 1 There are MCS1 and MCS2.

[0043] (3) Yeast transformation and positive verification The constructed SchCYP719G1b-pESC-His plasmid was transformed into Saccharomyces cerevisiae WAT11 competent cells. The transformation method is as follows: ① Place the carrier DNA in a 95℃ water bath or metal bath for 3 minutes, then quickly place it in an ice bath and let it stand for 3 minutes. Place it in a 95℃ water bath or metal bath for another 3 minutes, then quickly place it in an ice bath and let it stand for more than 3 minutes.

[0044] ②Take 100µL of WAT11 competent cells melted on ice, add about 2µg of pre-cooled recombinant plasmid, 10µL of Carrier DNA, and 500µL of PEG / LiAc in sequence, pipette and hit several times to mix, and place in a 30℃ water bath for 30min (invert 8 times every 15min to mix).

[0045] ③Put it in a 42℃ water bath for 15 min (invert 8 times at 7.5 min to mix well).

[0046] ④ Centrifuge at 5000rpm for 40s and discard the supernatant. Resuspend the cells in 400µL ddH2O and centrifuge at 5000rpm for 30s and discard the supernatant.

[0047] ⑤ Resuspend the bacteria in 50 µL ddH2O, spread on plates, and culture at 29°C for about 96 hours.

[0048] ⑥ Pick a single clone on the plate and perform positive verification using the alkaline lysis method to obtain the SchCYP719G1b-pESC-His-WAT11 positive clone.

[0049] Example 4 Peptides Sch Expression and enzyme activity analysis of CYP719G1b 1. Peptide Sch Expression of CYP719G1b (1) Select the positive clone of strain SchCYP719G1b-pESC-His-WAT11 and inoculate it into 50 mL His-deficient medium (prepared with 8 g His minus media and 1 L H2O, sterilized by high temperature and high pressure), and culture it in a shaking incubator at 29°C and 220 rpm for 2 days.

[0050] (2) The cultured bacterial liquid was centrifuged to collect the cells, and the cells were resuspended in 400 mL of YPL medium containing 2% (v / v) galactose, and cultured in a shaking incubator at 29°C and 220 rpm for 16 h to induce the expression of the target protein; the formula of YPL medium is (1 L): 10 g peptide and 10 g yeast extraction, dissolved in water and fixed to 1 L, and sterilized by high temperature and high pressure.

[0051] (3) Harvesting bacteria: Centrifuge the bacterial solution at 5,000 rpm for 5 min and discard the supernatant.

[0052] (4) Washing: Add 40 mL of TEK washing solution to the centrifuge tube to resuspend the cells, place on ice for 5 min, collect the cells by centrifugation at 4,000 × g for 10 min at 4°C, resuspend the cells with 200 mL of TESB solution, and place on ice for 10 min; TEK washing solution (350 mL): Dissolve 2.60925 g of KCl in TE solution and make up to 350 mL with TE solution, and mix well; TESB solution (1 L): Dissolve 109.3 g of sorbitol in TE solution and make up to 1 L with TE solution, and mix well; TE solution (1 L) contains 50 mL of 1 M Tris-HCl solution and 3.2 mL of 0.5 M EDTA solution, make up to 1 L with ddH2O, and mix well.

[0053] (5) Lysis: Disrupt the bacterial solution using a homogenizer for 6 min and centrifuge at 12,000 × g for 15 min at 4°C to collect the supernatant.

[0054] (6) Sedimentation: Add 40 g PEG4,000 and 3.52 g NaCl to the supernatant and shake to completely dissolve.

[0055] (7) Incubate on ice for 15 min. Centrifuge at 12,000 × g for 15 min at 4°C and discard the supernatant.

[0056] (8) The precipitate is resuspended in 6 mL of TEG solution to obtain the extracted microsomal protein. The TEG solution (50 mL) contains 40 mL of TEG solution and 10 mL of glycerol. The extracted microsomal protein contains polypeptides. Sch The total protein of CYP719G1b. Currently, the common method for CYP450 verification is to extract yeast microsomal protein for verification.

[0057] 2. Peptide Sch Verification of CYP719G1b enzyme activity Peptides Sch The in vitro enzyme activity function of CYP719G1b was identified, and a control group was set up. The microsomal protein in the control group was the microsomal protein extracted with an empty vector by the same steps as above, the substrate was progomisin, and the enzyme activity reaction system was shown in Table 6.

[0058] Table 6 Enzyme activity reaction system (250µL)

[0059] Note: The amounts of reduced coenzyme II, flavin adenine dinucleotide, flavin mononucleotide, glucose-6-phosphate, and substrate indicate the final concentrations in the enzyme activity reaction system. The amount of microsomal protein (200 μL) indicates that 200 μL of the protein solution obtained by resuspending with TEG solution in the above steps is taken for functional verification.

[0060] The above components were mixed and reacted at 29°C overnight. 200 μL of ethyl acetate was added to extract the reaction solution. The supernatant was taken after centrifugation at 12,000 rpm for 5 min. The extraction was repeated once. After evaporating the organic solvent, the solution was redissolved in chromatographic methanol and the enzyme activity reaction was analyzed by LC-MS.

[0061] The results are as follows Figure 2 As shown, compared with the blank control, the peptide Sch CYP719G1b can catalyze the CC coupling of the benzyl ring of the dibenzylbutane lignan pre-gomisin to form the biphenylcyclooctene lignan gomisin J. The product was confirmed by comparison with the standard. The reaction process is as follows Figure 3 shown.

[0062] Example 5 Encoding polypeptide Sch The nucleotide sequence of CYP719G1b (SEQ ID No. 2) was designed for site-directed mutation, and the expression extraction and enzyme activity function verification of the mutated sequence were performed using the methods of Examples 2 to 4. The enzyme activity function test results were as follows: Figure 4 As shown, two mutants with increased activity, R218K and E468D, were obtained after mutation, while the activities of the remaining mutants remained unchanged or decreased.

[0063] Figure 4 WT means wild type, i.e. polypeptide Sch CYP719G1b; I359A represents a polypeptide Sch The isoleucine at position 359 of CYP719G1b was mutated to alanine; Y49A indicates polypeptide Sch The tyrosine at position 49 of CYP719G1b was mutated to alanine; R218A represents polypeptide Sch The arginine at position 218 of CYP719G1b was mutated to alanine; I222A indicates polypeptide Sch The isoleucine at position 222 of CYP719G1b was mutated to alanine; K476A represents polypeptide Sch Lysine at position 476 of CYP719G1b was mutated to alanine; R218K represents polypeptide Sch The arginine at position 218 of CYP719G1b was mutated to lysine; E468D represents a polypeptide Sch CYP719G1b glutamic acid at position 468 mutated to aspartic acid; D224E represents a polypeptide SchThe aspartic acid at position 224 of CYP719G1b was mutated to glutamic acid; F469Y represents polypeptide Sch The phenylalanine at position 469 of CYP719G1b mutated to tyrosine; R242S indicates polypeptide Sch The arginine at position 242 of CYP719G1b was mutated to serine; T109L represents polypeptide Sch The threonine at position 109 of CYP719G1b was mutated to leucine; D224S indicates polypeptide Sch The aspartic acid at position 224 of CYP719G1b was mutated to serine.

[0064] Peptides Sch Amino acid sequence of CYP719G1b: MEGQWVVAAVLLLIGLILLRLWSKPEEKVAKWPPGPPKLPILGNFHQLYNGGNLLHHTIGKLVQEYGPIMTIWLGGWKPTIVVSDYELVREVLVTKAAEFGARNFPFSTRYLSADLKTVASAD YGPYWHTLRKGLQNSSLNPVNISAQLRMLESEFGVIIDTLASEALKNGGVVSPLEHMKLASIRLASRLCFGPKFNDEEFVQQLYALIQIIVRQTRESGISDLVPFTRHIPILGRFFHQRLHAM KLVGDLIRPYLHLSEPSTFLYFLKSQNLPEDILVANVFELFLLSADSSANSTMWGLGYLIHNQGIQQKLYDEIKSICGEGRVKVTAQDLNKMEYIHVVAKETLRMKPIAPLGIPRKATEDNTL MGYKIHKGTAIVLNLYQVLYDGKVWEEPERFMPERFLAGYGDEDRVMAMEKSFVSFGGGRRICPGIDLAKLMIPLALANLVGAYHWYGVDEESPDMTEEFVFGLMMKTPFSARIMPRKSG(SEQ ID No.1); Encoded polypeptide Sch Gene sequence of CYP719G1b:

[0065] Although the specific implementation of the present invention is described in detail in conjunction with the embodiments, it should not be understood as limiting the scope of protection of this patent. Within the scope described in the claims, various modifications and variations that can be made by those skilled in the art without creative work still fall within the scope of protection of this patent.

Claims

1. A polypeptide that catalyzes the cyclization of lignans, characterized in that The polypeptide catalyzing the cyclization of lignans is a polypeptide Sch CYP719G1b, whose amino acid sequence is selected from: (1) the amino acid sequence shown in SEQ ID No. 1; (2) an amino acid sequence produced by substitution, deletion and / or addition of one or more amino acids to the amino acid sequence shown in SEQ ID No. 1, and having the same catalytic activity as (1); or (3) A truncated form of (1) or (2) having the same catalytic activity as (1).

2. The gene encoding the polypeptide that catalyzes the cyclization of lignans according to claim 1, characterized in that The polypeptide Sch The coding gene sequence of CYP719G1b is selected from: (1) the nucleotide sequence shown in SEQ ID NO.2; (2) a nucleotide sequence having more than 80% homology with the nucleotide sequence shown in SEQ ID NO.2, and the encoded polypeptide has the same catalytic activity as the polypeptide encoded by (1); or (3) A truncated form of (1) or (2) having the same catalytic activity as the polypeptide encoded by (1).

3. An expression vector containing the coding gene according to claim 2.

4. A recombinant microorganism or transgenic cell line containing the encoding gene according to claim 2.

5. The method for preparing the polypeptide that catalyzes the cyclization of lignans according to claim 1, characterized in that: The method comprises the following steps: introducing the coding gene according to claim 2 into a receptor cell, expressing and extracting the polypeptide, and obtaining the product.

6. The use of the polypeptide for catalyzing the cyclization of lignans according to claim 1, characterized in that: Peptide Sch CYP719G1b is used to catalyze the cyclization of dibenzylbutane-type lignans.

7. The use according to claim 6, characterized in that: Peptides Sch CYP719G1b catalyzes the CC coupling in biphenylcyclooctene-type lignans.

8. The use according to claim 7, characterized in that: Peptides Sch CYP719G1b catalyzes the conversion of progomisin to gomisin J.

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