1,2-Pentanediol biosynthesis method and recombinant microorganism for producing 1,2-pentanediol

Abstract

The present invention belongs to the field of metabolic engineering, and specifically relates to a 1,2-pentanediol biosynthesis method and a recombinant microorganism for producing 1,2-pentanediol. The synthesis method of the present invention includes the following enzymes: threonine dehydratase, α-isopropylmalate synthase, α-isopropylmalate isomerase, β-isopropylmalate dehydrogenase, 2-oxidocarboxylic acid reductase, acyl CoA transferase, acyl CoA reductase, aldehyde reductase, cinnamic acid acylase, etc., and the above-mentioned enzyme gene encoding plasmid is used to obtain a recombinant microorganism, and the recombinant microorganism is fermented and cultured with an organic carbon source such as glucose to obtain high-yield 1,2-pentanediol. The present invention provides a biological method for synthesizing 1,2-pentanediol, and can significantly increase the yield of 1,2-pentanediol, and has broad application prospects.

Description

Technical Field

[0001] The invention belongs to the field of metabolic engineering, and specifically relates to a 1,2-pentanediol biosynthesis method and a recombinant microorganism for producing 1,2-pentanediol. Background Art

[0002] As a new type of moisturizer, 1,2-pentanediol is now widely used in the cosmetics field, such as skin creams, eye creams, skin lotions, baby care products, sunscreen products and other skin care products. At the same time, it has a certain antiseptic effect, reducing allergies caused by preservatives; it can also improve the water resistance of sunscreen product formulas; and dissolve insoluble active ingredients.

[0003] At present, the production methods of 1,2-pentanediol are mainly divided into two types according to the source of raw materials: (1) Fossil energy sources: 1-pentene is epoxidized to form pentyl oxide, and then hydrolyzed to produce 1,2-pentanediol;

[0004] (2) Biomass source: Lignin degradation produces furfural and furfuryl alcohol, which are then hydrogenated by chemical catalysts to form 1,2-pentanediol.

[0005] Based on the chemical conversion process of fossil raw materials, this production method not only leads to large amounts of CO 2 Emissions are also affected by fluctuations in the price of fossil raw materials, which directly affects the production cost of 1,2-pentanediol. Therefore, it is necessary to develop a method for producing 1,2-pentanediol based on microbial fermentation. However, biosynthetic methods still face many challenges, including the lack of corresponding biosynthetic pathways in nature, the lack of efficient biocatalytic elements, poor adaptation to chassis cells, and production capacity generally not meeting industrial requirements. Therefore, how to design and develop biosynthetic pathways that do not exist in nature, and use this pathway for optimization to obtain high-yield 1,2-pentanediol, is a problem that needs to be solved. Summary of the invention

[0006] In view of the problems in the prior art, the present invention provides a 1,2-pentanediol biosynthesis method and a recombinant microorganism for producing 1,2-pentanediol.

[0007] A method for biosynthesizing 1,2-pentanediol, starting from 2-oxopentanoic acid, via 2-hydroxypentanoic acid and 2-hydroxyvaleraldehyde intermediates, to synthesize 1,2-pentanediol in three steps:

[0008] Step 1: Reduction of 2-oxopentanoic acid to 2-hydroxypentanoic acid;

[0009] Step 2: Reduction of 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde;

[0010] Step 3: Reduction of 2-hydroxypentanal to 1,2-pentanediol.

[0011] Preferably, step 2 can obtain 2-hydroxyvaleraldehyde by a one-step reaction or a two-step reaction; the one-step reaction is: reducing 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde; the two-step reaction is: (1) converting 2-hydroxyvaleric acid to 2-hydroxyvaleryl-CoA; (2) reducing 2-hydroxyvaleryl-CoA to 2-hydroxyvaleraldehyde;

[0012] The enzyme used to reduce 2-oxopentanoic acid to 2-hydroxypentanoic acid is 2-oxocarboxylic acid reductase;

[0013] And / or, the enzyme used to convert 2-hydroxyvaleric acid into 2-hydroxyvaleryl-CoA is acyl CoA transferase or acyl CoA ligase;

[0014] and / or, the enzyme used to reduce 2-hydroxyvaleryl-CoA to 2-hydroxyvaleraldehyde is acyl-CoA reductase;

[0015] And / or, the enzyme used to reduce 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde is carboxylic acid reductase;

[0016] And / or, the enzyme used to reduce 2-hydroxyvaleraldehyde to 1,2-pentanediol is aldehyde reductase;

[0017] And / or, cinnamate acylase is overexpressed in the step of converting 2-hydroxyvalerate into 2-hydroxyvaleryl-CoA.

[0018] Preferably, the method is reduction and conversion under conditions including at least one of a crude enzyme solution, a combination of purified enzymes, an immobilized enzyme, and whole cells.

[0019] Preferably, the whole cell is a recombinant microorganism; the starting strain of the recombinant microorganism is selected from at least one of Escherichia coli, Corynebacterium glutamicum, Bacillus subtilis, Saccharomyces cerevisiae and Yarrowia lipolytica.

[0020] Preferably, the 2-oxocarboxylate reductase is selected from the following gene encodings: one or more of ldh, ldhA, panE, ara, ara1, ara2, araDH, YGL039W, adh, and CXQ87_003259;

[0021] And / or, the acyl-CoA transferase or acyl-CoA ligase is selected from one or more of the following genes encoding: fldA, hadA, pct, Cat2, carB, 4CL, PaaK, Cat1, PGN_0725, PGN_1341, PGN_1888, fadD;

[0022] And / or, the acyl-CoA reductase is selected from one or more of the following genes encoding: maqu2220, maqu2507, bld, ald, pdcD, pduP, adhE, FAR, acr, hom, fadB, fadJ, hbd, phaB, phbB, mdh, ldh;

[0023] And / or, the carboxylate reductase is encoded by a CAR gene;

[0024] And / or, the aldehyde reductase is selected from the following gene encoding: one or more of yqhD, ADH2, adh, bdh, yahK, adhA, 4-hbd, dhaT, mmsb, P84067, adhI, alrA, sucD, and bld;

[0025] And / or, the cinnamate acylase is selected from one or more of the following gene encodings: Pt4CL, At4CL2, At4CL5, Os4CL, Pc4CL.

[0026] The above method also includes, based on this method, continuing to increase the yield of 1,2-pentanediol by adding precursors, cofactors or increasing the concentrations of precursors and cofactors; the precursor is at least one of threonine and pyruvate, and the cofactor is NAD(P)H, NAD(P) + , ATP or at least one of them.

[0027] Preferably, a recombinant microorganism expressing ldh, CAR, and yqhD genes is taken, cultured in a culture medium, induced to express the recombinant microorganism, 2-oxopentanoic acid is added, and fermentation is continued to obtain 1,2-pentanediol;

[0028] Alternatively, a recombinant microorganism expressing ldh, fldA, maqu2507, and yqhD genes is taken, cultured in a culture medium, the expression of the recombinant microorganism is induced, 2-oxopentanoic acid is added, and fermentation is continued to obtain 1,2-pentanediol;

[0029] Alternatively, a recombinant microorganism expressing ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, 2-oxopentanoic acid is added, and fermentation is continued to obtain 1,2-pentanediol;

[0030] Alternatively, a recombinant microorganism expressing ilvA, leuABCD, ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, threonine is added, and fermentation is continued to obtain 1,2-pentanediol;

[0031] Alternatively, a recombinant microorganism expressing cimA, leuABCD, ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, pyruvic acid is added, and fermentation is continued to obtain 1,2-pentanediol;

[0032] Alternatively, a recombinant microorganism expressing ilvA, leuABCD, ldh, fldA, maqu2507, yqhD, Pc4CL, and zwf genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, threonine is added, and fermentation is continued to obtain 1,2-pentanediol;

[0033] The present invention also provides 1,2-pentanediol prepared according to the above method.

[0034] The present invention also provides a recombinant microorganism, wherein the recombinant microorganism overexpresses genes for 2-oxocarboxylic acid reductase, carboxylic acid reductase, acyl CoA reductase, acyl CoA transferase, acyl CoA ligase, cinnamate acylase and / or aldehyde reductase;

[0035] Preferably, the starting strain of the recombinant microorganism is selected from one or more of Escherichia coli, Corynebacterium glutamicum, Bacillus subtilis, Saccharomyces cerevisiae and Yarrowia lipolytica.

[0036] Preferably, the 2-oxocarboxylate reductase is selected from the following gene encodings: one or more of ldh, ldhA, panE, ara, ara1, ara2, araDH, YGL039W, adh, and CXQ87_003259;

[0037] And / or, the acyl-CoA transferase or acyl-CoA ligase is selected from one or more of the following genes encoding: fldA, hadA, pct, Cat2, carB, 4CL, PaaK, Cat1, PGN_0725, PGN_1341, PGN_1888, fadD;

[0038] And / or, the acyl-CoA reductase is selected from one or more of the following genes encoding: maqu2220, maqu2507, bld, ald, pdcD, pduP, adhE, FAR, acr, hom, fadB, fadJ, hbd, phaB, phbB, mdh, ldh;

[0039] And / or, the carboxylate reductase is encoded by a CAR gene;

[0040] And / or, the aldehyde reductase is selected from the following gene encoding: one or more of yqhD, ADH2, adh, bdh, yahK, adhA, 4-hbd, dhaT, mmsb, P84067, adhI, alrA, sucD, and bld;

[0041] And / or, the cinnamate acylase is selected from one or more of the following gene encodings: Pt4CL, At4CL2, At4CL5, Os4CL, Pc4CL.

[0042] The raw material 2-oxopentanoic acid used in the method of the present invention can be prepared according to the following method:

[0043] Step a, threonine is deaminated and dehydrated to generate 2-oxobutyric acid, or pyruvic acid is condensed with one molecule of acetyl-CoA to generate 2-oxobutyric acid;

[0044] Step b, 2-oxobutyric acid and acetyl coenzyme A are condensed, dehydrated, added, oxidized, and decarboxylated to obtain.

[0045] Preferably, threonine dehydratase is used to convert threonine into 2-oxobutyrate; and / or 2-methylmalate synthetase is used to condense pyruvate with one molecule of acetyl-CoA to generate 2-oxobutyrate;

[0046] And / or, a combination of α-isopropylmalate synthase, α-isopropylmalate isomerase and β-isopropylmalate dehydrogenase is used to convert 2-oxobutyrate into 2-oxopentanoate.

[0047] In the recombinant microorganism, threonine dehydratase is encoded by the ilvA gene; and / or, 2-methylmalate synthase is encoded by the cimA gene; and / or, a combination of α-isopropylmalate synthase, α-isopropylmalate isomerase and β-isopropylmalate dehydrogenase is encoded by the leuABCD gene.

[0048] In the fermentation method of the present invention, the culture conditions are: 37°C, 200 rpm, adding an inducer to the culture medium, fermenting for 48 hours, and obtaining the target product. The pH of the culture medium used for the culture is 6.8, and the formula of the culture medium includes 20 g / L glucose, 12.8 g / L Na 2 HPO 4 7H 2 O, 3g / LKH 2 PO 4 , 0.5g / LNaCl, 1g / LNH 4 Cl, 0.5 g / LMgSO 4 , 0.5g / L cinnamic acid, 5g / L yeast powder, 0.01g / L CaCl 2, 100 mg / L ampicillin, 35 mg / L chloramphenicol; the inducer used in the fermentation is selected from isopropyl-β-D-thiogalactoside.

[0049] The present invention provides a recombinant microorganism, which overexpresses genes of 2-oxocarboxylic acid reductase, carboxylic acid reductase, and aldehyde reductase;

[0050] or, overexpressing genes of 2-oxocarboxylate reductase, acyl-CoA transferase or acyl-CoA ligase, acyl-CoA reductase, aldehyde reductase;

[0051] or, overexpressing genes for 2-oxocarboxylic acid reductase, acyl-CoA transferase or acyl-CoA ligase, acyl-CoA reductase, cinnamate acylase, aldehyde reductase;

[0052] or, overexpressing genes of threonine dehydratase, α-isopropylmalate synthetase, α-isopropylmalate isomerase, β-isopropylmalate dehydrogenase, 2-oxocarboxylate reductase, acyl CoA transferase or acyl CoA ligase, acyl CoA reductase, cinnamate acylase, aldehyde reductase;

[0053] or, overexpressing genes of 2-methylmalate synthase, α-isopropylmalate synthase, α-isopropylmalate isomerase, β-isopropylmalate dehydrogenase, 2-oxocarboxylate reductase, acyl CoA transferase or acyl CoA ligase, acyl CoA reductase, cinnamate acylase, aldehyde reductase;

[0054] or, overexpression of genes for threonine dehydratase, α-isopropylmalate synthetase, α-isopropylmalate isomerase, β-isopropylmalate dehydrogenase, 2-oxocarboxylic acid reductase, acyl CoA transferase or acyl CoA ligase, acyl CoA reductase, cinnamate acylase, aldehyde reductase, glucose-6-phosphate-dehydrogenase;

[0055] Preferably, the starting strain of the recombinant microorganism is selected from Escherichia coli, Corynebacterium glutamicum, Bacillus subtilis, Saccharomyces cerevisiae or Yarrowia lipolytica.

[0056] The invention provides a 1,2-pentanediol biosynthesis method and adopts a recombinant microorganism encoded by an enzyme gene to produce 1,2-pentanediol according to the synthesis method; wherein threonine is used as a raw material, and the enzymes involved include threonine dehydratase (ilvA), α-isopropylmalate synthase (leuA), α-isopropylmalate isomerase (leuC, D), β-isopropylmalate dehydrogenase (leuB), lactate dehydrogenase (ldh), acyl CoA transferase (fldA), cinnamic acylase (Pc4CL), acyl CoA reductase (maqu2507), aldehyde reductase (yqhD), and glucose-6-phosphate dehydrogenase (zwf); the enzyme gene encoding plasmid is used, the plasmid is transferred into an Escherichia coli strain, the related enzymes from threonine to 1,2-pentanediol are overexpressed, and organic carbon sources such as glucose are used as substrates for fermentation culture, so as to obtain 1,2-pentanediol with a yield of 3.2 g / L. The present invention introduces a cinnamic acylase gene (Pc4CL) and a glucose-6-phosphate dehydrogenase gene (zwf) through optimization experiments, thereby significantly improving the yield of 1,2-pentanediol, which has important reference significance for promoting continuous innovation and development in the field of biosynthesis and meeting the requirements of the chemical industry for safety, economy, environmental protection and sustainable development.

[0057] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0058] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 Pathway 1 from 2-oxopentanoate to 1,2-pentanediol: direct reduction by carboxylic acid reductase.

[0060] Figure 2 Pathway 2 from 2-oxopentanoate to 1,2-pentanediol: through the formation of acyl-CoA and then reduction.

[0061] Figure 3 It is the pathway from threonine to 2-oxopentanoate.

[0062] Figure 4 It is the pathway from pyruvate to 2-oxopentanoate.

[0063] Figure 5 The plasmid is pTrc99a-yqhD-sfp-car.

[0064] Figure 6 It is plasmid pSTV28-ldh.

[0065] Figure 7 The plasmid is pTrc99a-yqhD-maqu2507-pct.

[0066] Figure 8 The plasmid is pTrc99a-yqhD-maqu2507-fldA.

[0067] Fig. 9 The plasmid is pTrc99a-yqhD-maqu2507-Ptrc-fldA-4CL.

[0068] Fig.10 It is plasmid pSTV28-leuABCD-Ptrc-ilvA-ldh.

[0069] Fig.11 It is plasmid pSTV28-leuABCD-Ptrc-cimA-ldh.

[0070] Fig.12 It is plasmid pSTV28-leuABCD-Ptrc-ilvA-ldh-zwf.

[0071] Fig.13 Comparison of the conversion efficiency of different acyl-CoA reductases.

[0072] Fig.14 Comparison of the conversion efficiency of different CoA transferases.

[0073] Fig.15 Comparison of different 4CL conversion efficiencies.

[0074] Fig.16 To increase the effect of NADPH supply on 1,2-pentanediol production. DETAILED DESCRIPTION

[0075] The present invention relates to the following nucleotide sequence information:

[0076] Nucleotide sequence of lactate dehydrogenase (ldh) (SEQ ID NO.1)

[0077] ATGACTAAAATTGCCATGTATAATGTCAGCCCGATCGAAGTGCCTTACATTGAAGACT

[0078] GGGCTAAGAAAAACGATGTCGAAATTAAGACGACCGACCAGGCCTTGACCAGCGC

[0079] GACTGTAGATTTAGCTGAAGGGTGCAGCAGCGTTTCCCTTAAGCCACTTGGCCCGGT

[0080] TGACGAAGAAGTTGTTTACCAAAAATTGAGCGAATACGGGGTCAAGTGCATCGGCC

[0081] TCAGAATCGTTGGCTTCAACACCATCAACTTCGACTGGACCAAGAAGTACAACTTG

[0082] CTGGTCACCAACGTTCCGGTCTACTCCCCGCGGGCGATCGCTGAAATGACGGTTACC

[0083] CAGGCCATGTACCTCCTGCGCAAGATCGGGGAATTCCGCTACCGGATGGACCATGAC

[0084] CATGACTTTACCTGGCCAAGTAACTTGATCAGCAATGAAATCTACAACTTGACTGTC

[0085] GGCTTGATCGGGGTCGGTCACATCGGCAGCGCCGTGGCAGAAATCTTCTCAGCCATG

[0086] GGGGCCAAGGTCATCGCCTATGACGTGGCTTACAACCCGGAATTTGAACCATTTTTG

[0087] ACCTACACCGACTTTGACACGGTCTTGAAAGAAGCTGACATCGTCTCCCTCCACACT

[0088] CCCCTCTTCCCATCAACGGAAAACATGATCGGGGAAAAGCAGCTGAAGAAAATGAA

[0089] GAAGTCTGCCTACTTGATCAACTGTGCCCGGGGCGAATTAGTCGACACTGGAGCCTT

[0090] GATCAAGGCCTTGCAGGATGGCGAAATCGCCGGTGCCGGCCTGGACACTTTGGCTG

[0091] GGGAATCCAGCTACTTTGGCCACACTGGCCTGACGGACAGCGAAATCCCAGAAGAC

[0092] TACAAGACCCTGGCCAAGATGCCAAACGTTGTCATTACTCCGCACTCAGCCTTCTAC

[0093] ACGGAAACTTCCATCCGCAACATGGTGCAGATCTGCTTGACTGACCAATTGACCATC

[0094] GCCAAGGGCGGCCGGCCAAGAAGCATCGTTAACCTGTAA

[0095] Nucleotide sequence of carboxylic acid reductase (car) (SEQ ID NO.2)

[0096] ATGACCGTGACCAACGAAACCAACCCACAGCAGGAGCAGCTATCCCGCCGTATTGA

[0097] AAGTCTGCGCGAAAGCGATCCGCAGTTCCGGGCGGCCCAGCCCGACCCGGCGGTCG

[0098] CCGAACAGGTGCTGCGCCCGGGCCTGCATCTTTCTGAAGCCATTGCGGCGTTGATGA

[0099] CTGGATACGCTGAGCGCCCGGCGCTCGGTGAGCGCGCACGCGAGTTGGTCACCGAC

[0100] CAGGATGGCCGCACCACGCTGCGCCTGTTGCCACGCTTCGACACCACCACATACGG

[0101] CGAATTATGGTCCCGCACAACATCAGTCGCCGCTGCATGGCACCACGACGCCGCCCA

[0102] CCCGGTTAAGGCCGGCGATCTGGTGGCCACCCTGGGATTCACCAGCATCGACTACAC

[0103] CGTGCTGGATCTGGCGATCATGATCCTCGGTGGCGTGGCGGTTCCGCTACAGACCAG

[0104] CGCCCCGGCTTCGCAGTGGACGACCATTCTGGCCGAAGCGGAACCCAACACTCTTG

[0105] CGGTAAGCATCGAATTGATCGGCGCTGCAATGGAATCTGTGCGGGCCACGCCTTCCA

[0106] TCAAGCAGGTCGTCGTGTTCGACTACACCCCCGAGGTCGATGATCAACGGGAGGCA

[0107] TTCGAGGCAGCAAGCACACAACTCGCCGGCACCGGCATCGCCATTGAGACCCTCGA

[0108] TGCCGTCATCGCCCGCGGCGCCGCACTTCCGGCCGCACCGCTCTACGCACCATCGGC

[0109] CGGCGACGATCCGCTGGCGCTGCTCATCTACACCTCCGGCAGCACCGGGGCTCCAA

[0110] AGGGCGCCATGCACAGCGAAAACATCGTGCGCCGCTGGTGGATTCGTGAGGACGTC

[0111] ATGGCCGGCACCGAGAACCTGCCCATGATCGGGCTGAACTTCATGCCGATGAGTCAC

[0112] ATCATGGGACGCGGCACCCTCACCTCCACCCTGTCTACCGGTGGAACCGGATACTTC

[0113] GCGGCGTCCAGTGACATGTCAACGCTCTTCGAGGACATGGAGCTGATCCGCCCGAC

[0114] GGCCCTGGCCTTGGTTCCACGCGTGTGCGACATGGTGTTCCAGCGATTCCAGACCGA

[0115] GGTGGACCGGCGTCTGGCGAGCAGCGACACCGCCAGTGCCGAGGCCGTTGCGGCC

[0116] GAGGTCAAGGCCGATATCCGTGACAACCTCTTCGGTGGCCGCGTATCGGCGGTCATG

[0117] GTCGGTTCTGCTCCGTTGTCCGAGGAGCTGGGTGAGTTCATCGAATCCTGCTTCGAG

[0118] CTGAATCTGACCGATGGCTACGGCTCCACCGAAGCCGGCATGGTGTTCCGCGACGG

[0119] CATCGTGCAACGCCCGCCGGTCATTGACTACAAGCTGGTTGACGTGCCCGAACTGG

[0120] GCTACTTCTCCACCGACAAGCCGCACCCGCGCGGTGAGCTGCTGCTGAAGACCGAC

[0121] GGCATGTTCCTCGGGTACTACAAACGCCCCGAGGTGACTGCCGGCGTCTTCGACGC

[0122] GGACGGTTTTTACATGACCGGCGACATCGTCGCCGAGCTGGCCCACGACAACATCG

[0123] AGATCATCGATCGCCGCAACAACGTGCTCAAACTCTCACAGGGAGAGTTTGTCGCG

[0124] GTCGCCACCTTGGAGGCCGAGTACGCCAATAGCCCTGTGGTGCACCAGATCTACGTC

[0125] TACGGCAGCAGCGAACGGTCCTACCTGCTAGCAGTCGTGGTGCCGACGCCGGAGGC

[0126] CGTGGCCGCCGCCAAGGGCGACGCGGCGGCACTCAAGACGACCATCGCGGACTCG

[0127] CTGCAGGACATTGCCAAGGAGATCCAGCTGCAGTCCTACGAAGTCCCCCGTGACTT

[0128] CATCATCGAACCGCAGCCATTCACCCAGGGCAACGGCCTGCTGACGGGTATCGCCA

[0129] AGCTGGCGCGTCCGAACCTGAAGGCGCACTATGGACCGCGGCTGGAGCAGATGTAC

[0130] GCCGAAATCGCCGAGCAGCAGGCTGCCGAGCTTCGGGCGTTGCACGGAGTGGACC

[0131] CAGACAAGCCCGCGCTGGAAACGGTCCTCAAGGCGGCGCAGGCCCTGCTCGGCGT

[0132] CTCGTCGGCCGAACTGGCCGCGGACGCGCATTTCACCGATCTAGGTGGCGATTCGCT

[0133] GTCCGCACTGTCCTTCTCGGATCTGCTGCGCGATATCTTCGCGGTCGAAGTACCGGT

[0134] CGGAGTCATCGTCAGTGCCGCAAACGATCTCAGCGGTGTTGCGAAATTTGTTGATGA

[0135] ACAACGCTATTCGGGCGGGACGCGGCCGACCGCGGAGACGGTGCACGGCGCCGGG

[0136] CATACGGAGATCCGGGCCGCGGACCTGACCCTGGATAAGTTCATCGACGAGGCCAC

[0137] CCTGCATGCGGCACCGTCGCTTCCGAAGGCCGTCGGGATCCCACACACCGTCCTGC

[0138] TCACCGGGTCCAACGGCTACCTGGGCCACTACCTGGCACTGGAATGGCTTGAGCGC

[0139] CTGGACAAGACAGAAGGCAAGCTGATCGCCATCGTCCGCGGTAAGAATGCCGAGGC

[0140] CGCCTACCGCCGCCTCGAGGAAGCCTTCGACACCGGCGACACGCAGCTGTTGGCGC

[0141] ACTTCCGGTCGCTGGCCGACAAGCACCTCGAAGTACTGGCCGGCGATATCGGCGAC

[0142] CCCAACCTTGGCCTGGATGCCGACACCTGGCAGCGCCTGGCCGACACCGTCGACGT

[0143] CATCGTGCACCCCGCCGCCCTGGTCAACCACGTACTGCCCTACAGCCAGCTGTTCGG

[0144] ACCGAATGTCGTCGGCACCGCCGAGATCATCAAGCTGGCCATCACTACCAAGATCAA

[0145] GCCGGTCACCTACCTGTCCACGGTCGCGGTCGCGGCATATGTCGATCCGACGACATT

[0146] CGACGAAGAGTCCGATATCCGGCTCATCAGCGCGGTGCGTCCCGTGGACGAGCTGT

[0147] ACGCGAACGGCTACGGCAACAGCAAGTGGGCCGGCGAGGTACTGCTGCGCGAAGC

[0148] CCACGATCTGTGCGGACTACCCGTCGCGGTCTTCCGCTCCGACATGATCTTGGCCCA

[0149] CAGCCGCTACACCGGACAGCTCAACGTGCCCGACCAGTTCACCCGACTAATCCTCA

[0150] GCCTCATCGCCACCGGAATCGCACCCGGCTCCTTCTACCAAGCACACGCCACCGGC

[0151] GAACGCCCACTCGCCCACTACGACGGGCTACCCGGTGACTTCACCGCCGAGGCGAT

[0152] CACCACGTTGGGCACCCAGGTGGTCGACAGCTACGAGACCTACGACTGCGTGAACC

[0153] CGCATGCAGACGGAGTCTCGCTGGACAACTTCGTCGACTGGCTCATCGAAGCCGGC

[0154] TACCCCATCGCACGCATCGACAACTACACCGAATGGTTCACCCGCTTCGACACCGCC

[0155] ATCCGAAGCCTCCCCGAAAAACAGAAACAACACTCCCTACTACCACTGCTCCACGC

[0156] ATTCGAACAGCCGTCCGCCGCCGAGAACCACGGCGTCGTCCCGGCAAAGCGTTTCC

[0157] AGCACGCTGTGCAGGCCGCCGGAATCGGTCCGGCCGGGCAAGACGGCACTACCGA

[0158] CATTCCCCACCTGTCGCGGCGGCTGATCGTGAAATACGCCAAGGACCTCGAACAGC

[0159] TCGGACTCCTATGA

[0160] Nucleotide sequence of CoA transferase (fldA) (SEQ ID NO.3)

[0161] ATGGAAAACAACGCCAATATGTTCAGCGGTGTGAAAGTTATTGAACTGGCAAACTTT

[0162] ATTGCAGCACCGGCAGCAGGTCGTTTCTTTGCAGATGGTGGTGCAGAAGTTATCAAA

[0163] ATTGAAAGTCCGGCAGGCGATCCGCTGCGTTATACCGCACCGAGCGAAGGTCGTCC

[0164] GCTGAGCCAAGAAGAAAATACCACCTACGATCTGGAAAACGCAAACAAAAAGGCC

[0165] ATTGTGCTGAATCTGAAAAGCGAGAAAGGTAAAAAGATCCTGCACGAGATGCTGAA

[0166] AGAAACCGATATTCTGCTGACCAATTGGCGTACCAAAGCACTGGTTAAACAAGGTCT

[0167] GGATTATGAAACCCTGAAAGAGAAATATCCGAAACTGGTTTTTGCACAGATTACCGG

[0168] TTATGGTGAAAAAGGTCCGGATAAAGATTTACCGGGTTTTGATTATACCGCCTTTTTC

[0169] GCACGTGGTGGTGTTAGCGGCACCCTGTATGAAAAAGGCACCGTTCCGCCTAATGTT

[0170] GTTCCTGGTCTGGGTGATCATCAGGCAGGTATGTTTCTGGCAGCCGGTATGGCAGGC

[0171] GCACTGTATAAAGCCAAAACCACCGGTAAAGGTGATAAAGTTACCGTTAGCCTGATG

[0172] CATAGCGCAATGTATGGTCTGGGCATTATGATTCAGGCAGCACAGTATACCGGTCATG

[0173] GCCTGGTTTATCCGATTAACCGTAATGAAACCCCGAATCCGTTTATCGTTAGCTACAA

[0174] AAGCAAAGACGATTATTTCGTTCAGGTTTGCATGCCTCCGTATGATGTGTTTTATGAT

[0175] CGTTTTATGACCAGCCTGGGTCGTGAGGATCTGGTTGGTGATGAACGTTATAACAAA

[0176] ATCGAGAACCTGAAAGATGGTCGTGCCAAAGAAGTGTATAAAATCATCGAAGAACA

[0177] AATGGTGACCAAGACCAAAGATGAGTGGGATAAAATCTTTCGCGAAGCCGATATTCC

[0178] GTTTGCCATTGCACAGACCTGGGAAGATTTACTGCAGGATGAACAGGCATGGGCAA

[0179] ATGATTATCTGTACAAAATGAAATACCCGACCGGTAATGAACGTGCACTGGTTCGTCT

[0180] GCCGGTGTTTTTCAAAGAAGCAGGTCTGCCGGAATATAAACAGAGTCCGCAGATTG

[0181] CCGAAAATACAGCCGAAGTTTTAAAAGAGATGGGCTATACCGAAGAGGATATCCAA

[0182] GAACTGGAAAAGAACAAAGATATCATGGTGCGCAAAGAAAAATAA

[0183] Nucleotide sequence of acyl-CoA reductase (maqu2507) (SEQ ID NO.4)

[0184] ATGAACTATTTTCTGACCGGTGGCACCGGCTTTATTGGTCGTTTTCTGGTTGAAAAAC

[0185] TGCTGGCACGTGGTGGTACAGTTTATGTTCTGGTTCGTGAACAGAGCCAGGATAAAC

[0186] TGGAACGTCTGCGTGAACGTTGGGGTGCAGATGATAAACAGGTTAAAGCAGTTATT

[0187] GGTGACCTGACCAGCAAAAATCTGGGTATTGATGCAAAAACCCTGAAAAGCCTGAA

[0188] AGGCAATATCGATCATGTTTTTCATCTGGCAGCCGTGTATGATATGGGTGCCGATGAA

[0189] GAAGCACAGGCAGCAACCAATATTGAAGGCACCCGTGCAGCCGTTCAGGCAGCCG

[0190] AAGCAATGGGTGCAAAACATTTTCATCATGTTAGCAGCATTGCAGCAGCAGGTCTGT

[0191] TTAAAGGTATTTTTCGCGAAGATATGTTTGAAGAGGCCGAAAAACTGGATCATCCGT

[0192] ATCTGCGTACCAAACATGAAAGCGAAAAAGTTGTTCGTGAAGAATGCAAAGTGCCG

[0193] TTTCGTATTTATCGTCCTGGTATGGTTATTGGTCATAGCGAAACCGGTGAAATGGATA

[0194] AAGTTGATGGTCCGTACTACTTCTTTAAAATGATCCAGAAAATTCGTCATGCACTGCC

[0195] GCAGTGGGTTCCGACCATTGGTATTGAAGGTGGTCGTCTGAATATTGTTCCGGTTGAT

[0196] TTTGTTGTTGATGCCCTGGATCACATTGCACATCTGGAAGGTGAAGATGGTAATTGTT

[0197] TTCACCTGGTTGATAGCGATCCGTATAAAGTTGGTGAAATCCTGAACATCTTTTGCGA

[0198] AGCAGGTCATGCACCGCGTATGGGTATGCGTATTGATAGCCGTATGTTTGGTTTTATC

[0199] CCTCCGTTTATTCGTCAGAGCATTAAAAACCTGCCTCCGGTTAAACGTATTACCGGTG

[0200] CACTGCTGGATGACATGGGTATTCCGCCTAGCGTTATGAGCTTTATTAACTATCCGAC

[0201] ACGTTTTGATACCCGTGAGCTGGAACGTGTTCTGAAAGGTACAGATATTGAAGTTCC

[0202] GCGTCTGCCGAGCTATGCACCGGTTATTTGGGATTATTGGGAACGTAATCTGGACCC

[0203] GGACCTGTTCAAAGATCGTACCTTAAAAGGCACCGTTGAAGGTAAAGTTTGTGTTGT

[0204] GACCGGTGCAACCAGCGGTATTGGTCTGGCAACCGCAGAGAAACTGGCCGAAGCG

[0205] GGTGCAATTCTGGTGATTGGTGCACGTACCAAAGAAACCCTGGATGAAGTTGCAGC

[0206] AAGCCTGGAAGCAAAAGGTGGTAATGTTCATGCATATCAGTGCGATTTCAGCGATAT

[0207] GGATGATTGCGATCGTTTTGTTAAAACCGTGCTGGATAATCATGGTCATGTTGATGTG

[0208] CTGGTTAATAATGCAGGTCGTAGCATTCGTCGTAGCCTGGCACTGAGCTTTGATCGTT

[0209] TTCATGATTTTGAACGTACCATGCAGCTGAATTATTTCGGTAGCGTTCGTCTGATTATG

[0210] GGCTTTGCACCGGCAATGCTGGAACGCCGTCGTGGTCATGTGGTGAATATTAGCTCA

[0211] ATTGGTGTTCTGACCAACGCACCGCGTTTTAGCGCCTATGTGAGCAGCAAAAGCGC

[0212] ACTGGATGCATTTAGCCGTTGTGCAGCAGCCGAATGGTCAGATCGTAATGTTACCTTT

[0213] ACCACCATTAATATGCCGCTGGTTAAAACCCCGATGATTGCACCGACCAAAATCTATG

[0214] ATAGCGTTCCGACACTGACACCGGATGAAGCAGCCCAGATGGTTGCAGATGCAATT

[0215] GTGTATCGTCCGAAACGTATTGCAACCCGTCTGGGTGTTTTTGCACAGGTTCTGCAT

[0216] GCACTGGCACCGAAAATGGGTGAAATTATCATGAATACCGGCTATCGTATGTTTCCGG

[0217] ATAGTCCGGCAGCCGCAGGTAGCAAAAGTGGTGAAAAACCGAAAGTTAGCACCGA

[0218] ACAGGTTGCATTTGCAGCAATTATGCGTGGTATTTACTGGTAA

[0219] Nucleotide sequence of cinnamic acid acyltransferase (Pc4CL) (SEQ ID NO.5)

[0220] ATGGGTGATTGTGTTGCACCGAAAGAGGATCTGATTTTTCGTAGCAAACTGCCGGAT

[0221] ATCTATATCCCGAAACATCTGCCGCTGCATACCTATTGTTTTGAGAATATTAGCAAAGT

[0222] GGGCGACAAAAGCTGTCTGATTAATGGTGCAACCGGTGAAACCTTTACCTATAGCCA

[0223] GGTTGAACTGCTGAGCCGTAAAGTTGCAAGCGGTCTGAATAAACTGGGTATTCAGC

[0224] AGGGTGATACCATTATGCTGCTGCTGCCGAATAGTCCGGAATATTTCTTTGCATTTCT

[0225] GGGTGCAAGCTATCGTGGTGCCATTAGCACCATGGCAAATCCGTTTTTCACCAGCGC

[0226] AGAAGTTATTAAACAGCTGAAAGCAAGCCAGGCCAAACTGATTATTACCCAGGCAT

[0227] GTTATGTGGACAAAGTGAAAGATTATGCAGCCGAGAAAAACATCCAGATCATTTGCA

[0228] TTGATGATGCACCGCAGGATTGTCTGCATTTTAGTAAACTGATGGAAGCCGATGAAA

[0229] GCGAAATGCCGGAAGTTGTGATTAATAGTGATGATGTTGTTGCACTGCCGTATAGCA

[0230] GCGGTACAACCGGTCTGCCGAAAGGTGTTATGCTGACCCATAAAGGTCTGGTTACCA

[0231] GCGTTGCACAGCAGGTTGATGGTGATAATCCGAATCTGTATATGCATAGCGAAGATGT

[0232] GATGATTTGTATCCTGCCGCTGTTTCATATCTATAGCCTGAATGCAGTTCTGTGTTGTG

[0233] GTCTGCGTGCCGGTGTTACCATTCTGATCATGCAGAAATTTGATATCGTGCCGTTTCT

[0234] GGAACTGATCCAGAAATATAAGGTTACCATTGGTCCGTTTGTTCCGCCTATTGTTCTG

[0235] GCAATTGCAAAAAGTCCGGTGGTGGATAAATATGATCTGAGCAGCGTTCGTACCGTT

[0236] ATGAGCGGTGCAGCTCCGCTGGGTAAAGAACTGGAAGATGCAGTTCGTGCAAAATT

[0237] TCCGAATGCCAAATTAGGTCAAGGTTATGGTATGACCGAAGCAGGTCCGGTTCTGGC

[0238] CATGTGTCTGGCATTTGCCAAAGAACCGTATGAAATCAAAAGCGGTGCATGTGGCAC

[0239] CGTTGTTCGTAATGCCGAAATGAAAATTGTTGATCCGGAAACCAATGCAAGCCTGCC

[0240] TCGTAATCAGCGTGGTGAAATTTGTATTCGTGGTGACCAGATTATGAAAGGCTATCTG

[0241] AATGATCCTGAAAGCACCCGTACCACCATTGATGAAGAAGGTTGGCTGCATACAGGT

[0242] GATATTGGCTTTATCGATGATGATGACGAACTGTTTATTGTGGATCGCCTGAAAGAGA

[0243] TCATCAAATACAAAGGTTTTCAGGTGGCACCAGCAGAGCTGGAAGCACTGCTGCTG

[0244] ACACATCCGACCATTAGTGATGCAGCAGTTGTTCCGATGATTGATGAAAAAGCCGGT

[0245] GAAGTTCCGGTTGCATTTGTTGTGCGTACCAATGGTTTTACCACCACGGAAGAAGAA

[0246] ATCAAACAGTTTGTTAGCAAACAGGTGGTGTTCTACAAACGTATTTTTCGCGTTTTCT

[0247] TTGTGGATGCCATTCCGAAAAGCCCGAGCGGTAAAATTCTGCGTAAAGACCTGCGT

[0248] GCACGTATTGCCAGCGGTGATCTGCCTAAATAA

[0249] Nucleotide sequence of leuABCD (SEQ ID NO.6)

[0250] ATGAGCCAGCAAGTCATTATTTTCGATACCACATTGCGCGACGGTGAACAGGCGTTA

[0251] CAGGCAAGCTTGAGTGTGAAAGAAAAACTGCAAATTGCGCTGGCCCTTGAGCGTAT

[0252] GGGTGTTGACGTGATGGAAGTCGGTTTCCCCGTCTCTTCGCCGGGCGATTTTGAATC

[0253] GGTGCAAACCATCGCCCGCCAGGTTAAAAACAGCCGCGTATGTGCGTTAGCTCGCT

[0254] GCGTGGAAAAAGATATCGACGTGGCGGCCGAATCCCTGAAAGTCGCCGAAGCCTTC

[0255] CGTATTCATACCTTTATTGCCACTTCGCCAATGCACATCGCCACCAAGCTGCGCAGCA

[0256] CGCTGGACGAGGTGATCGAACGCGCTATCTATATGGTGAAACGCGCCCGTAATTACA

[0257] CCGATGATGTTGAATTTTCTTGCGAAGATGCCGGGCGTACACCCATTGCCGATCTGG

[0258] CGCGAGTGGTCGAAGCGGCGATTAATGCCGGTGCCACCACCATCAACATTCCGGAC

[0259] ACCGTGGGCTACACCATGCCGTTTGAGTTCGCCGGAATCATCAGCGGCCTGTATGAA

[0260] CGCGTGCCTAACATCGACAAAGCCATTATCTCCGTACATACCCACGACGATTTGGGC

[0261] CTGGCGGTCGGAAACTCACTGGCGGCGGTACATGCCGGTGCACGCCAGGTGGAAG

[0262] GCGCAATGAACGGGATCGGCGAGCGTGCCGGAAACTGTTCCCTGGAAGAAGTCATC

[0263] ATGGCGATCAAAGTTCGTAAGGATATTCTCAACGTCCACACCGCCATTAATCACCAG

[0264] GAGATATGGCGCACCAGCCAGTTAGTTAGCCAGATTTGTAATATGCCGATCCCGGCA

[0265] AACAAAGCCATTGTTGGCAGCGGCGCATTCGCACACTCCTCCGGTATACACCAGGAT

[0266] GGCGTGCTGAAAAACCGCGAAAACTACGAAATCATGACACCAGAATCTATTGGTCT

[0267] GAACCAAATCCAGCTGAATCTGACCTCTCGTTCGGGGCGTGCGGCGGTGAAACATC

[0268] GCATGGATGAGATGGGGTATAAAGAAAGTGAATATAATTTAGACAATTTGTACGATGC

[0269] TTTCCTGAAGCTGGCGGACAAAAAAGGTCAGGTGTTTGATTACGATCTGGAGGCGC

[0270] TGGCCTTCATCGGTAAGCAGCAAGAAGAGCCGGAGCATTTCCGTCTGGATTACTTCA

[0271] GCGTGCAGTCTGGCTCTAACGATATCGCCACCGCCGCCGTCAAACTGGCCTGTGGCG

[0272] AAGAAGTCAAAGCAGAAGCCGCCAACGGTAACGGTCCGGTCGATGCCGTCTATCAG

[0273] GCAATTAACCGCATCACTGAATATAACGTCGAACTGGTGAAATACAGCCTGACCGCC

[0274] AAAGGCCACGGTAAAGATGCGCTGGGTCAGGTGGATATCGTCGCTAACTACAACGG

[0275] TCGCCGCTTCCACGGCGTCGGCCTGGCTACCGATATTGTCGAGTCATCTGCCAAAGC

[0276] CATGGTGCACGTTCTGAACAATATCTGGCGTGCCGCAGAAGTCGAAAAAGAGTTGC

[0277] AACGCAAAGCTCAACACAACGAAAACAACAAGGAAACCGTGTGATGTCGAAGAAT

[0278] TACCATATTGCCGTATTGCCGGGGGACGGTATTGGTCCGGAAGTGATGACCCAGGCG

[0279] CTGAAAGTGCTGGATGCCGTGCGCAACCGCTTTGCGATGCGCATCACCACCAGCCAT

[0280] TACGATGTAGGCGGCGCAGCCATTGATAACCACGGGCAACCACTGCCGCCTGCGAC

[0281] GGTTGAAGGTTGTGAGCAAGCCGATGCCGTGCTGTTTGGCTCGGTAGGCGGCCCGA

[0282] AGTGGGAACATTTACCACCAGACCAGCAACCAGAACGCGGCGCGCTGCTGCCTCTG

[0283] CGTAAGCACTTCAAATTATTCAGCAACCTGCGCCCGGCAAAACTGTATCAGGGGCTG

[0284] GAAGCATTCTGTCCGCTGCGTGCAGACATTGCCGCAAACGGCTTCGACATCCTGTGT

[0285] GTGCGCGAACTGACCGGCGGCATCTATTTCGGTCAGCCAAAAGGCCGCGAAGGTAG

[0286] CGGACAATATGAAAAAGCCTTTGATACCGAGGTGTATCACCGTTTTGAGATCGAACG

[0287] TATCCGCCCGCATCGCGTTTGAATCTGCTCGCAAGCGTCGCCACAAAGTGACGTCGAT

[0288] CGATAAAGCCAACGTGCTGCAATCCTCTATTTTATGGCGGGAGATCGTTAACGAGAT

[0289] CGCCACGGAATACCCGGATGTCGAACTGGCGCATATGTACATCGACAACGCCACCAT

[0290] GCAGCTGATTAAAGATCCATCACAGTTTGACGTTCTGCTGTGCTCCAACCTGTTTGG

[0291] CGACATTCTGTCTGACGAGTGCGCAATGATCACTGGCTCGATGGGGATGTTGCCTTC

[0292] CGCCAGCCTGAACGAGCAAGGTTTTGGACTGTATGAACCGGCGGGCGGCTCGGCAC

[0293] CAGATATCGCAGGCAAAAACATCGCCAACCCGATTGCACAAATCCTTTCGCTGGCAC

[0294] TGCTGCTGCGTTACAGCCTGGATGCCGATGATGCGGCTTGCGCCATTGAACGCGCCA

[0295] TTAACCGCGCATTAGAAGAAGGCATTCGCACCGGGGATTTAGCCCGTGGCGCTGCCG

[0296] CCGTTAGTACCGATGAAATGGGCGATATCATTGCCCGCTATGTAGCAGAAGGGGTGT

[0297] AATCATGGCTAAGACGTTATACGAAAAATTGTTCGACGCTCACGTTGTGTACGAAGC

[0298] CGAAAACGAAACCCCACTGTTATATATCGACCGCCACCTGGTGCATGAAGTGACCTC

[0299] ACCGCAGGCGTTCGATGGTCTGCGCGCCCACGGTCGCCCGGTACGTCAGCCGGGCA

[0300] AAACCTTCGCTACCATGGATCACAACGTCTCTACCCAGACCAAAGACATTAATGCCT

[0301] GCGGTGAAATGGCGCGTATCCAGATGCAGGAACTGATCAAAAACTGCAAAGAATTT

[0302] GGCGTCGAACTGTATGACCTGAATCACCCGTATCAGGGGATCGTCCACGTAATGGGG

[0303] CCGGAACAGGGCGTCACCTTGCCGGGGATGACCATTGTCTGCGGCGACTCGCATAC

[0304] CGCCACCCACGGCGCGTTTGGCGCACTGGCCTTTGGTATCGGCACTTCCGAAGTTGA

[0305] ACACGTACTGGCAACGCAAACCCTGAAACAGGGCCGCGCAAAAACCATGAAAATT

[0306] GAAGTCCAGGGCAAAGCCGCGCCGGGCATTACCGCAAAAGATATCGTGCTGGCAAT

[0307] TATCGGTAAAACCGGTAGCGCAGGCGGCACCGGGCATGTGGTGGAGTTTTGCGGCG

[0308] AAGCAATCCGTGATTTAAGCATGGAAGGTCGTATGACCCTGTGCAATATGGCAATCG

[0309] AAATGGGCGCAAAAGCCGGTCTGGTTGCACCGGACGAAACCACCTTTAACTATGTC

[0310] AAAGGCCGTCTGCATGCGCCGAAAGGCAAAGATTTCGACGACGCCGTTGCCTACTG

[0311] GAAAACCCTGCAAACCGACGAAGGCGCAACTTTCGATACCGTTGTCACTCTGCAAG

[0312] CAGAAGAAATTTCACCGCAGGTCACCTGGGGCACCAATCCCGGCCAGGTGATTTCC

[0313] GTGAACGACAATATTCCCGATCCGGCTTCGTTTGCCGATCCGGTTGAACGCGCGTCG

[0314] GCAGAAAAAGCGCTGGCCTATATGGGGCTGAAACCGGGTATTCCGCTGACCGAAGT

[0315] GGCTATCGACAAAGTGTTTATCGGTTCCTGTACCAACTCGCGCATTGAAGATTTACGC

[0316] GCGGCAGCGGAGATCGCCAAAGGGCGAAAAGTCGCGCCAGGCGTGCAGGCACTGG

[0317] TGGTTCCCGGCTCTGGCCCGGTAAAAGCCCAGGCGGAAGCGGAAGGTCTGGATAAA

[0318] ATCTTTATTGAAGCCGGTTTTGAATGGCGCTTGCCTGGCTGCTCAATGTGTCTGGCGA

[0319] TGAACAACGACCGTCTGAATCCGGGCGAACGTTGTGCCTCCACCAGCAACCGTAAC

[0320] TTTGAAGGCCGCCAGGGGCGCGGCGGGCGCACGCATCTGGTCAGCCCGGCAATGGC

[0321] TGCCGCTGCTGCTGTGACCGGACATTTCGCCGACATTCGCAACATTAAATAAGGAGC

[0322] ACACCATGGCAGAGAAATTTATCAAACACACAGGCCTGGTGGTTCCGCTGGATGCC

[0323] GCCAATGTCGATACCGATGCAATCATCCCGAAACAGTTTTTGCAGAAAGTGACCCGT

[0324] ACGGGTTTTGGCGCGCATCTGTTTAACGACTGGCGTTTTCTGGATGAAAAAGGCCAA

[0325] CAGCCAAACCCGGACTTCGTGCTGAACTTCCCGCAGTATCAGGGCGCTTCCATTTTG

[0326] CTGGCACGAGAAAACTTCGGCTGTGGCTCTTCGCGTGAGCACGCGCCCTGGGCATT

[0327] GACCGACTACGGTTTTAAAGTGGTGATTGCGCCGAGTTTTGCTGACATCTTCTACGG

[0328] CAATAGCTTTAACAACCAGCTGCTGCCGGTGAAATTAAGCGATGCAGAAGTGGACG

[0329] AACTGTTTGCGCTGGTGAAAGCTAATCCGGGGATCCATTTCGACGTGGATCTGGAAG

[0330] CGCAAGAGGTGAAAGCGGGAGAGAAAACCTATCGCTTTACCATCGATGCCTTCCGC

[0331] CGCCACTGCATGATGAACGGTCTGGACAGTATTGGGCTTACCTTGCAGCACGACGAC

[0332] GCCATTGCCGCTTATGAAGCAAAACAACCTGCGTTTATGAATTAA

[0333] Nucleotide sequence of 2-methylmalate synthase (cimA) (SEQ ID NO.7)

[0334] ATGGCTGACTCGCAACCCCTGTCCGGTGCTCCGGAAGGTGCCGAATATTTAAGAGCA

[0335] GTGCTGCGCGCGCCGGTTTACGAGGCGGCGCAGGTTACGCCGCTACAAAAAATGGA

[0336] AAAACTGTCGTCGCGTCTTGATAACGTCATTCTGGTGAAGCGCGAAGATCGCCAGCC

[0337] AGTGCACAGCTTTAAGCTGCGCGGCGCATACGCCATGATGGCGGGCCTGACGGAAG

[0338] AACAGAAAGCGCACGGCGTGATCACTGCTTCTGCGGGTAACCACGCGCAGGGCGTC

[0339] GCGTTTTCTTCTGCGCGGTTAGGCGTGAAGGCCCTGATCGTTATGCCAACCGCCACC

[0340] GCCGACATCAAAGTCGACGCGGTGCGCGGCTTCGGCGGCGAAGTGCTGCTCCACGG

[0341] CGCGAACTTTGATGAAGCGAAAGCCAAAGCGATCGAACTGTCACAGCAGCAGGGG

[0342] TTCACCTGGGTGCCGCCGTTCGACCATCCGATGGTGATTGCCGGGCAAGGCACGCTG

[0343] GCGCTGGAACTGCTCCAGCAGGACGCCCATCTCGACCGCGTATTTGTGCCAGTCGG

[0344] CGGCGGCGGTCTGGCTGCTGGCGTGGCGGTGCTGATCAAACAACTGATGCCGCAAA

[0345] TCAAAGTGATCGCCGTAGAAGCGGAAGACTCCGCCTGCCTGAAAGCAGCGCTGGAT

[0346] GCGGGTCATCCGGTTGATCTGCCGCGCGTAGGGCTATTTGCTGAAGGCGTAGCGGTA

[0347] AAACGCATCGGTGACGAAACCTTCCGTTTATGCCAGGAGTATCTCGACGACATCATC

[0348] ACCGTCGATAGCGATGCGATCTGTGCGGCGATGAAGGATTTATTCGAAGATGTGCGC

[0349] GCGGTGGCGGAACCCTCTGGCGCGCTGGCGCTGGCGGGAATGAAAAAATATATCGC

[0350] CCTGCACAACATTCGCGGCGAACGGCTGGCGCATATTCTTTCCGGTGCCAACGTGAA

[0351] CTTCCACGGCCTGCGCTACGTCTCAGAACGCTGCGAACTGGGCGAACAGCGTGAAG

[0352] CGTTGTTGGCGGTGACCATTCCGGAAGAAAAAGGCAGCTTCCTCAAATTCTGCCAA

[0353] CTGCTTGGCGGGCGTTCGGTCACCGAGTTCAACTACCGTTTTGCCGATGCCAAAAAC

[0354] GCCTGCATCTTTGTCGGTGTGCGCCTGAGCCGCGGCCTCGAAGAGCGCAAAGAAAT

[0355] TTTGCAGATGCTCAACGACGGCGGCTACAGCGTGGTTGATCTCTCCGACGACGAAAT

[0356] GGCGAAGCTACACGTGCGCTATATGGTCGGCGGACGTCCATCGCATCCGTTGCAGGA

[0357] ACGCCTCTACAGCTTCGAATTCCCGGAATCACCGGGCGCGCTGCTGCGCTTCCTCAA

[0358] CACGCTGGGTACGTACTGGAACATTTCTTTGTTCCACTATCGCAGCCATGGCACCGA

[0359] CTACGGGCGCGTACTGGCGGCGTTCGAACTTGGCGACCATGAACCGGATTTCGAAA

[0360] CCCGGCTGAATGAGCTGGGCTACGATTGCCACGACGAAACCAATAACCCGGCGTTC

[0361] AGGTTCTTTTTGGCGGGTTAG

[0362] Nucleotide sequence of CoA transferase (hadA) (SEQ ID NO.8)

[0363] ATGCTGCTGGAAGGTGTTAAAGTTGTTGAACTGAGCAGCTTTATTGCAGCACCGTGT

[0364] TGTGCAAAAATGTTAGGTGATTGGGGTGCCGAAGTGATTAAAATCGAACCGATTGAA

[0365] GGTGATGGTATTCGTGTTATGGGTGGCACCTTTAAAAGTCCGGCAAGTGATGATGAA

[0366] AACCCGATGTTTGAACTGGAAAACGGTAATAAGAAAGGCGTGAGCATTAACGTGAA

[0367] AAGCAAAGAAGGTGTGGAAATCCTGCATAAACTGCTGAGCGAAGCAGATATTTTTG

[0368] TTACCAATGTTCGTGTTCAGGCCCTGGAAAAGATGGGTATTGCCTATGATCAGATCAA

[0369] AGACAAATATCCGGGTCTGATTTTTAGCCAGATTTTAGGCTATGGTGAAAAAGGTCC

[0370] GCTGAAAGATAAACCGGGTTTTGATTATACCGCATATTTTGCACGTGGTGGTGTTAGC

[0371] CAGAGCGTTATGGAAAAAGGCACCTCACCGGCAAATACCGCAGCAGGTTTTGGTGA

[0372] TCATTATGCAGGTCTGGCACTGGCAGCAGGTAGCCTGGCAGCACTGCATAAGAAAG

[0373] CACAGACCGGTAAAGGTGAACGTGTTACCGTTAGCCTGTTTCATACCGCAATTTATG

[0374] GTATGGGCACCATGATTACCACCGCACAGTATGGTAATGAAATGCCGCTGAGCCGTG

[0375] AAAATCCGAATAGTCCGCTGATGACCACCTATAAATGTAAAGATGGTCGTTGGATTC

[0376] AGCTGGCACTGATTCAGTATAACAAATGGCTGGGCAAATTCTGCAAAGTGATCAACC

[0377] GTGAGTATATCCTGGAAGATGATCGCTATAACAACATCGATAGCATGGTGAATCATGT

[0378] TGAGGATCTGGTTAAAATTGTTGGTGAAGCAATGCTTGAGAAAACCCTGGATGAATG

[0379] GTCAGCACTGTTAGAAGAAGCCGATCTGCCGTTTGAGAAAATTCAGAGCTGCGAGG

[0380] ATCTGCTGGATGATGAACAGGCATGGGCAAATGATTTTCTGTTCAAAAAGACCTATG

[0381] ATAGCGGTAATACCGGTGTTCTGGTTAATACACCGGTTATGTTTCGCAACGAAGGCAT

[0382] CAAAGAATACACACCGGCACCGAAAGTTGGTCAGCATACCGTTGAAGTTCTGAAAA

[0383] GCCTGGGTTATGATGAAGAGAAAATTAACAACTTCAAAGATAGCAAGGTGGTGCGC

[0384] TATTAA

[0385] Nucleotide sequence of CoA transferase (pct) (SEQ ID NO.9)

[0386] ATGCGCAAAGTGGAAATTATCACCGCAGAACAGGCAGCACAGCTGGTTAAAGATAA

[0387] TGATACCATTACCAGCATTGGCTTTGTTAGCAGCGCACATCCGGAAGCACTGACCAA

[0388] AGCACTGGAAAAACGCTTTCTGGATACCAATACTCCGCAGAATCTGACCTATATCTAT

[0389] GCAGGTAGCCAGGGTAAACGTGATGGTCGTGCAGCAGAACATCTGGCACATACCGG

[0390] TCTGCTGAAACGTGCAATTATTGGTCATTGGCAGACCGTTCCGGCAATTGGTAAACT

[0391] GGCAGTTGAAAACAAAATCGAGGCCTATAACTTTAGCCAGGGCACCCTGGTTCATTG

[0392] GTTTCGTGCACTGGCAGGTCATAAACTGGGTGTTTTTACCGATATTGGTCTGGAAAC

[0393] CTTTTTAGATCCGCGTCAGTTAGGTGGCAAACTGAATGATGTTACCAAAGAGGATCT

[0394] GGTCAAACTGATTGAAGTTGATGGTCATGAGCAGCTGTTTTATCCGACCTTTCCGGTT

[0395] AATGTTGCATTTCTGCGTGGCACCTATGCAGATGAAAGCGGCAATATTACCATGGATG

[0396] AAGAAATTGGTCCGTTTGAAAGCACCAGCGTTGCACAGGCCGTTCATAATTGTGGTG

[0397] GTAAAGTTGTTGTTCAGGTGAAAGATGTTGTTGCCCATGGTAGCCTTGATCCGCGTAT

[0398] GGTTAAAATTCCGGGTATCTATGTTGATTATGTTGTTGTTGCAGCACCGGAAGATCAT

[0399] CAGCAGACGTATGATTGTGAATATGATCCGAGCCTGAGCGGTGAACATCGTGCTCCG

[0400] GAAGGTGCAACCGATGCAGCACTGCCGATGAGCGCCAAAAAGATTATTGGCCGTCG

[0401] TGGTGCACTGGAACTGACCGAAAATGCAGTTGTGAATTTAGGTGTTGGTGCACCGG

[0402] AATATGTTGCAAGCGTTGCCGGTGAAGAAGGTATTGCAGATACAATTACCCTGACCG

[0403] TTGAAGGTGGTGCAATTGGTGGTGTTCCGCAAGGTGGCGCACGTTTTGGTAGCAGC

[0404] CGTAATGCAGATGCCATTATTGATCATACCTATCAGTTCGATTTCTATGATGGTGGTGG

[0405] TCTGGATATTGCATATCTGGGTTTAGCACAGTGTGATGGTAGCGGTAATATCAACGTT

[0406] AGCAAATTTGGCACCAATGTTGCAGGTTGCGGTGGTTTTCCGAATATTAGCCAGCAG

[0407] ACCCCGAATGTGTATTTTTGTGGCACCTTTACCGCAGGCGGTCTGAAAATTGCGGTT

[0408] GAAGATGGTAAAGTGAAAATCCTGCAAGAAGGCAAAGCCAAGAAATTTATCAAAGC

[0409] CGTTGATCAGATTACGTTCAATGGTAGCTATGCAGCACGTAATGGTAAACATGTTCTG

[0410] TATATTACCGAACGCTGCGTTTTTGAACTGACAAAAGAAGGTCTGAAACTGATCGAG

[0411] GTTGCACCGGGTATTGATATCGAAAAAGATATTCTGGCCCACATGGATTTCAAACCGA

[0412] TTATTGATAATCCGAAGCTGATGGATGCACGTCTGTTTCAGGATGGTCCGATGGGTCT

[0413] GAAGAAATAA

[0414] Nucleotide sequence of CoA transferase (Cat2) (SEQ ID NO.10)

[0415] ATGAAAGACGTATTAGCGGAATATGCCTCCCGAATTGTTTCGGCCGAAGAAGCCGTA

[0416] AAACATATCAAAAATGGAGAACGGGTAGCTTTGTCACATGCTGCCGGAGTTCCTCAG

[0417] AGTTGTGTTGATGCACTGGTACAACAGGCCGACCTTTTCCAGAATGTCGAAATTTAT

[0418] CACATGCTTTGTCTCGGCGAAGGAAAATATATGGCACCTGAAATGGCCCCTCACTTC

[0419] CGACACATAACCAATTTTGTAGGTGGTAATTCTCGTAAAGCAGTTGAGGAAAATAGA

[0420] GCCGACTTCATTCCGGTATTCTTTTATGAAGTGCCATCAATGATTCGCAAAGACATCC

[0421] TTCACATAGATGTCGCCATCGTTCAGCTTTTCAATGCCTGATGAGAATGGTTACTGTAG

[0422] TTTTGGAGTATCTTGCGATTATAGCAAACCGGCAGCAGAAAGCGCTCATTTAGTTATA

[0423] GGGGAATCAACCGTCAAATGCCATATGTACATGGCGACAACTTGATTCACATATCG

[0424] AAGTTGGATTACATCGTGATGGCAGACTACCCTATCTATTCTCTTGCAAAGCCCAAAA

[0425] TCGGAGAAGTAGAAGAAGCTATCGGGCGTAATTGTGCCGAGCTTATTGAAGATGGTG

[0426] CCACACTCCAACTCGGTATCGGCGCGATTCCTGATGCAGCCCTGTTTATTCCTCAAGG

[0427] ACAAAAAAGATCTGGGGATCCATACCGAGATGTTCTCCGATGGTGTTGTCGAATTAG

[0428] TTCGCAGTGGAGTAATTACAGGAAAGAAAAGACACTTCACCCCGGAAAGATGGTC

[0429] GCAACCTTCTTAATGGGAAGCGAAGACGTATATCATTTCATCGACAAAAATCCCGAT

[0430] GTAGAACTTTATCCGGTAGATTACGTCAATGATCCGCGAGTAATCCGCTCAAAATGATA

[0431] ATATGGTCAGCATCAATAGCTGTATCGAAATCGATCTTATGGGACAAGTCGTGTCCGA

[0432] ATGTATAGGAAGCAAGCAATTCAGCGGAACCGGCGGTCAAGTAGATTATGTTCGTGG

[0433] AGCAGCATGGTCTAAAAACGGCAAAAGCATCATGGCAATTCCCTCAACAGCCAAAA

[0434] ACGGTACTGCATCTCGAATTGTACCTATAATTGCAGAGGGAGCTGCTGTAACAACCC

[0435] TCCGCAACGAAGTCGATTACGTTGTAACCGAATACGGTATAGCACAACTCAAAGGAA

[0436] AGAGTTTGCGCCAGCGAGCAGAAGCTCTTATTGCCATAGCCCACCCGGATTTCAGAG

[0437] AGGAACTAACGAAACATCTCCGCAAACGTTTCGGATAA

[0438] Nucleotide sequence of CoA transferase (carB) (SEQ ID NO.11)

[0439] ATGGAACTGATTCGCCATACCATCAACAGCTTTTTCAATGAACGTGTTAAAAAGACC

[0440] CCGAACAAAGTGGCAATTGTGTTTGGTGATGATGCATATACCTGGCTGGAACTGGAT

[0441] CAGATTAGTGATGGTATTGCACTGGAACTGCTGAAAAGTGATGTTAAGAAAGGCACC

[0442] CATGTTGGTATTTGGAGCCTGAATAGTCCGCTGTTTATGCTGTATTTTCTGGCACTGG

[0443] CAAAAATTGGTGCAGTTACCGTTCTGATTAATACCCGTTGTCATCTGAGCGAACTGG

[0444] AAGATTTACTGGATTATGCAGATGTTACCGTGCTGTGTTATGGTGAAGATAGTAACAC

[0445] CCTGAAATTTCTGGAAACCATTGAACAGCTGGATCGTAAACGTCTGCTGCTGATGAA

[0446] AAAGATTATTGCCATTGGCAAGAACAAAAAGGATATTCCGGCACAGTGTAAAAAGA

[0447] AACTGATCGAAGGTGATGATCTGAAGCTGATTGGTAATGCCAAGAAAGATGTTTTTC

[0448] CGGATGATGTTCTGGGCATTCTGTTTACCAGCGGTACAACCAGCGCAGCAAAAGGT

[0449] GTTATGCTGAGCCATTTTGCAACCATTAACATTGCGATTGAAACCGTGACACAGATG

[0450] CATTGGAATGAGAATGATAAAAGCTGTCTGGGTATCCCGCTGTTTCATTGTTTTGGTC

[0451] TGAGCAGCGGTTTTATGGCAGGTATTTATGCCGGTATTGAAATCTATCTGCTGGAAAA

[0452] CTATCAGACCATTCGTGTTCTGAAATGCGTTGAAACCAATCAGTGCACCATTTTTAAC

[0453] GGTGTTCCGACCATGTTTATGGCCATGATTAAACATCCTCGCTTCAAAACCTATAATC

[0454] TGAGCCTGCTGAATAGCGGTATTATTGCCGGTAGCTATGTTAACGCAAATGATTATCT

[0455] GAGCATCTGCAAAGAACTGAAAATGGAAAAACTGCAAATGAGCTATGGTCAGACCG

[0456] AAGCAAGCCCGAGCATTACCTTTAGCGATTATGAAGATACCATCCTGGATAAAAGCC

[0457] GTAGCGTTGGTAAAGTTATTCCGCATGTTGATCTGAAAATTCTGGTGAGCAACCAGA

[0458] ACGAAATTTGCATCAAAGGCTATAATACCATGCTGGGCTATTACAAACTGGATGAGG

[0459] AAACCAAAAAGGCCATTGATAGCGAAGGTTGGCTGCATACCGGTGATCTGGGTTATC

[0460] TGGATGCAGATGGTAATCTGTGTATTACCGGTCGTCTGAAAGAAATGATTGTTCGTGG

[0461] TGGTGAAAACATCAGCCCGTATGAAATTGAAAACCAGATCAAAAACTTTCCGTATGT

[0462] GGCCGATGTTAAAGTGATTGGCATTGAAGAAGAAGTGCTGCAAGAAGAAATTGCAG

[0463] CCTGTATTGTTACCGATGGTCGTAAAAAGCTGAACGAAACCGAAATTCGTGAATTTC

[0464] TGAGTCATTATCTGGCAGCCTATAAAATCCCGAAATACTTCATCTTCCTGAAAAAGTT

[0465] TCCGCTGAATGCAAGCGGTAAAATCATTCTGGCCGAACTGAAAGATCAGATCAAGAT

[0466] GATTATCAACACCAACAAAAATTAA

[0467] Nucleotide sequence of acyl-CoA reductase (maqu2220) (SEQ ID NO.12)

[0468] ATGGCAATTCAGCAGGTTCATCATGCAGATACCAGCAGCAGCAAAGTTCTGGGTCAG

[0469] CTGCGTGGTAAACGTGTTCTGATTACCGGCACCACCGGTTTTCTGGGTAAAGTTGTT

[0470] CTGGAACGTCTGATTCGTGCAGTTCCGGATATTGGTGCCATTTATCTGCTGATTCGCG

[0471] GTAATAAACGTCATCCTGATGCACGTAGCCGTTTTCTGGAAGAAATTGCAACCAGCA

[0472] GCGTTTTTGATCGTCTGCGTGAAGCAGATAGCGAAGGTTTTGATGCATTCCTGGAAG

[0473] AACGTATTCATTGTGTTACCGGTGAAGTTACCGAAGCAGGTTTTGGTATTGGTCAAG

[0474] AAGATTATCGTAAACTGGCAACCGAACTGGATGCAGTGATTAATAGCGCAGCAAGCG

[0475] TTAATTTTCGTGAGGAACTGGATAAAGCACTGGCAATTAATACCCTGTGTCTGCGTAA

[0476] TATTGCAGGTATGGTTGATCTGAATCCGAAACTGGCAGTTCTGCAGGTTAGCACCTG

[0477] TTATGTTAATGGTATGAATAGCGGTCAGGTTACCGAAAGCGTTATTAAACCGGCAGGC

[0478] GAAGCAGTTCCGCGTAGTCCGGATGGTTTTTATGAAATTGAAGAACTGGTTCGTCTG

[0479] CTGCAGGATAAAATCGAAGATGTTCAGGCACGTTATAGCGGTAAAGTGCTGGAACG

[0480] CAAACTGGTGGATCTGGGTATTCGTGAAGCAAATCGTTATGGTTGGAGCGATACCTAT

[0481] ACCTTTACCAAATGGCTGGGTGAACAGCTGCTGATGAAAGCACTGAATGGTCGTAC

[0482] CCTGACCATTCTGCGTCCGAGCATTATTGAAAGCGCATTAGAAGAACCGGCACCTGG

[0483] TTGGATTGAAGGTGTTAAAGTTGCAGATGCAATTATTCTGGCATATGCGCGTGAAAA

[0484] AGTTACCCTGTTTCCGGGTAAACGTAGCGGTATTATTGATGTTATTCCTGTTGATCTGG

[0485] TGGCCAACAGCATTATTCTGAGCCTGGCCGAAGCACTGGGCGAACCGGGTCGTCGT

[0486] CGTATTTATCAGTGTTGTAGCGGTGGTGGTAATCCGATTAGCCTGGGTGAATTTATTG

[0487] ATCATCTGATGGCAGAAAGCAAAGCCAATTATGCAGCATATGATCACCTGTTTTATCG

[0488] TCAGCCGAGCAAACCGTTTCTGGCAGTTAATCGTGCACTGTTTGACCTGGTTATTAG

[0489] CGGTGTGCGTCTGCCGCTGAGTCTGACCGATCGTGTGCTGAAACTGCTGGGTAATAG

[0490] CCGTGATCTGAAAATGCTGCGCAATCTGGATACCACACAGAGCCTGGCAACCATTTT

[0491] TGGCTTTTATACCGCACCGGATTATATCTTTCGTAACGATGAACTGATGGCACTGGCC

[0492] AATCGTATGGGTGAAGTTGATAAAGGTCTGTTTCCAGTTGATGCCCGTCTGATTGATT

[0493] GGGAACTGTATCTGCGCAAAATTCATCTGGCAGGTCTGAATCGCTATGCACTGAAAG

[0494] AACGTAAAGTGTATAGCCTGAAAACCGCACGTCAGCGTAAAAAGGCAGCATAA

[0495] Nucleotide sequence of acyl-CoA reductase (bld) (SEQ ID NO. 13)

[0496] ATGATTAAAGACACGCTAGTTTCTATAACAAAAGATTTAAAATTAAAAACAAATGTT

[0497] GAAAATGCCAATCTAAAGAACTACAAGGATGATTCTTCATGTTTCGGAGTTTTCGAA

[0498] AATGTTGAAAATGCTATAAGCAATGCCGTACACGCACAAAAGATATTATCCCTTCATT

[0499] ATACAAAAGAACAAAGAGAAAAAATCATAACTGAGATAAGAAAGGCCGCATTAGAA

[0500] AATAAAGAGATTCTAGCTACAATGATTCTTGAAGAAACACATATGGGAAGATATGAA

[0501] GATAAAATATTAAAGCATGAATTAGTAGCTAAATACACTCCTGGGACAGAAGATTTAA

[0502] CTACTACTGCTTGGTCAGGAGATAACGGGCTTACAGTTGTAGAAATGTCTCCATATGG

[0503] CGTTATAGGTGCAATAACTCCTTCTACGAATCCAACTGAAACTGTAATATGTAATAGTA

[0504] TAGGCATGATAGCTGCTGGAAATACTGTAGTATTTAACGGACATCCAGGCGCTAAAA

[0505] AATGTGTTGCTTTTGCTGTCGAAATGATAAATAAAGCTATTATTTCATGTGGTGGTCCT

[0506] GAGAATTTAGTAACAACTATAAAAAATCCAACTATGGAATCTCTAGATGCAATTATTA

[0507] AGCATCCTTCAATAAAACTACTTTGCGGAACTGGAGGGCCAGGAATGGTAAAAACC

[0508] CTCTTAAATTCTGGTAAGAAAGCTATAGGTGCTGGTGCTGGAAATCCACCAGTTATTG

[0509] TAGATGATACTGCTGATATAGAAAAGGCTGGTAAGAGTATCATTGAAGGCTGTTCTTT

[0510] TGATAATAATTTACCTTGTATTGCAGAAAAAGAAGTATTTGTTTTTGAGAATGTTGCA

[0511] GATAATTTAATATCTAACATGCTAAAAAATAATGCTGTAATTATAAATGAAGATCAAGT

[0512] ATCAAAGTTAATAGATTTAGTATTACAAAAAAATAATGAAACTCAAGAATACTCTATA

[0513] AATAAGAAATGGGTCGGAAAAGATGCAAAATTATTCTTAGATGAAATAGATGTTGAG

[0514] TCTCCTTCAAATGTTAAATGCATAATCTGCGAAGTGAGTGCAAGTCATCCATTTGTTA

[0515] TGACAGAACTCATGATGCCAATATTACCAATTGTAAGAGTTAAAGATATAGATGAAGC

[0516] TATTGAATATGCAAAAATAGCAGAACAAAATAGAAAACATAGTGCCTATATTTATTCA

[0517] AAAAATATAGACAACCTAAATAGGTTTGAAAGAGAAATCGATACTACTATCTTTGTAA

[0518] AGAATGCTAAATCTTTTGCCGGTGTTGGTTATGAAGCAGAAGGCTTTACAACTTTCA

[0519] CTATTGCTGGATCCACTGGTGAAGGAATAACTTCTGCAAGAAATTTTACAAGACAAA

[0520] GAAGATGTGTACTCGCCGGTTAA

[0521] Nucleotide sequence of acyl-CoA reductase (ald) (SEQ ID NO.14)

[0522] ATGAACAAAGATACCCTGATTCCGACCACCAAAGACCTGAAACTGAAAACCAATGT

[0523] GGAAAACATCAACCTGAAGAACTACAAAGATAACAGCAGCTGTTTTGGCGTGTTTG

[0524] AAAATGTTGAAAACGCCATTAATAGCGCAGTTCATGCACAGAAAATTCTGAGCCTGC

[0525] ATTATACCAAAGAACAGCGCGAAAAGATTATTACCGAAATTCGTAAAGCAGCCCTGG

[0526] AAAATAAAGAAGTTCTGGCAACCATGATTCTGGAAGAAACCCACATGGGTCGTTATG

[0527] AAGATAAAATCCTGAAACATGAACTGGTGGCCAAATATACACCGGGTACAGAGGATC

[0528] TGACCACCACCGCATGGTCAGGTGATAATGGTCTGACCGTTGTTGAAATGAGCCCGT

[0529] ATGGTGTTATTGGTGCAATTACCCCGAGCACCAATCCGACCGAAACCGTTATTTGTAA

[0530] TAGCATTGGTATGATTGCAGCCGGTAATGCCGTTGTTTTTAATGGTCATCCGGGTGCA

[0531] AAGAAATGTGTTGCATTTGCCATTGAGATGATCAACAAAGCCATTATTAGCTGTGGT

[0532] GGTCCGGAAAATCTGGTTACCACCATCAAAAATCCGACCATGGAAAGCCTGGATGC

[0533] CATTATCAAACATCCGCTGATTAAACTGCTGTGTGGCACCGGTGGTCCTGGTATGGTT

[0534] AAAACCCTGCTGAATAGCGGTAAAAAGGCAATTGGTGCCGGTGCAGGTAATCCGCC

[0535] TGTTATTGTTGATGATACCGCAGATATTGAGAAAGCCGGTAAAAGCATTATTGAAGGC

[0536] TGCAGCTTTGATAATAATCTGCCGTGTATTGCCGAAAAAGAGGTGTTTGTTTTTGAAA

[0537] ACGTTGCCGATGATCTGATCAGCAACATGCTGAAAAATAACGCCGTGATCATTAACG

[0538] AAGATCAGGTGAGCAAACTGATTGATCTGGTGCTGCAGAAAAACAATGAAACCCAA

[0539] GAGTACTTTATCAACAAAAAGTGGGTTGGCAAAGACGCCAAACTGTTTAGTGATGA

[0540] AATTGATGTTGAAAGCCCGAGCAACATTAAATGCATTGTTTGTGAAGTGAATGCCAA

[0541] CCATCCGTTTGTTATGACCGAACTGATGATGCCGATTCTGCCGATTGTTCGTGTTAAA

[0542] GATATTGATGAGGCCGTGAAATATACCAAAATTGCCGAACAGAATCGTAAACACAGC

[0543] GCCTATATCTACAGCAAAAACATCGATAATCTGAACCGCTTTGAACGCGAAATTGATA

[0544] CCACCATTTTTGTCAAAAACGCCAAGAGCTTTGCCGGTGTTGGTTATGAAGCAGAA

[0545] GGTTTTACCACCTTTACCATTGCAGGTAGCACCGGTGAAGGTATTACCAGCGCACGT

[0546] AATTTTACCCGTCAGCGTCGTTGTGTTCTGGCAGGTTAA

[0547] Nucleotide sequence of acyl-CoA reductase (pduP) (SEQ ID NO. 15)

[0548] ATGAATACAGCAGAACTGGAAACCCTTATCCGCACCATCCTCAGTGAAAAGCTCGC

[0549] GCCGACGCCCCCTGCCCCTCAGCAAGAGCAGGGCATTTTCTGCGATGTCGGCAGCG

[0550] CCATCGACGCCGCTCATCAGGCTTTTCTCCGCTATCAGCAGTGTCCGCTAAAAACCC

[0551] GCAGCGCCATTATCAGCGCCCTGCGGGAGACGCTGGCCCCCGAGCTGGCGACGCTG

[0552] GCGGAAGAGAGCGCCACGGAAACCGGCATGGGCAACAAAGAAGATAAATATCTGA

[0553] AAAATAAAGCCGCTCTTGAAAACACGCCGGGCATAGAGGATCTCACTACCAGCGCC

[0554] CTCACCGGCGATGGCGGGATGGTGCTGTTTGAGTACTCGCCGTTCGGGGTTATTGGC

[0555] GCCGTGGCGCCCAGCACCAACCCAACGGAAACCATTATCAACAACAGTATCAGCAT

[0556] GCTGGCGGCGGGTAACAGCGTCTATTTCAGCCCCCATCCCGGCGCGAAAAAGGTCT

[0557] CGTTGAAGCTTATCGCCAGGATCGAAGAGATCGCCTACCGCTGCAGCGGGATCCGTA

[0558] ACCTGGTGGTGACCGTTGCCGAGCCGACCTTTGAAGCCACCCAGCAAATGATGTCC

[0559] CACCCGCTGATTGCCGTTCTGGCTATCACCGGCGGCCCTGGCATTGTGGCGATGGGC

[0560] ATGAAAAGCGGTAAAAAAGTGATCGGCGCTGGCGCCGGCAATCCGCCGTGCATCGT

[0561] TGATGAAACCGCCGATCTCGTCAAAGCCGCCGAAGATATCATCAGCGGCGCCGCCTT

[0562] CGATTACAACCTGCCCTGTATCGCCGAAAAAAGCCTGATCGTCGTCGCCTCCGTCGC

[0563] TGACCGCCTGATCCAGCAGATGCAGGATTTTGACGCGCTGCTGTTGAGCCGACAGG

[0564] AGGCCGATACCCTGCGTGCCGTCTGCCTGCCCGACGGCGCGGCGAATAAAAAACTG

[0565] GTCGGTAAAAGCCCGGCTGCGCTGCTGGCGGCGGCGGGTCTCGCCGTTCCGCCTCG

[0566] CCCCCCTCGCCTGCTGATAGCCGAGGTGGAGGCGAACGACCCCTGGGTGACCTGCG

[0567] AGCAGCTGATGCCGGTGCTGCCGATCGTCAGGGTCGCCGACTTTGACAGCGCCCTG

[0568] GCGCTGGCCCTGCGCGTTGAGGAGGGTCTGCACCACACCGCCATTATGCACTCGCA

[0569] GAATGTCTCGCGGCTCAATCTGGCGGCACGCACGCTGCAGACCTCCATTTTTGTCAA

[0570] AAATGGCCCGTCTTACGCGGGAATCGGCGTCGGCGGCGAAGGGTTTACCACCTTCA

[0571] CCATCGCCACGCCAACCGGAGAAGGCACCACCTCCGCGCGGACGTTCGCCCGCCTG

[0572] CGGCGCTGCGTGTTGACCAACGGTTTTTCCATTCGCTAA

[0573] Nucleotide sequence of cinnamic acid acyltransferase (Pt4CL) (SEQ ID NO.16)

[0574] ATGGCCAACGGTATTAAGAAAGTGGAACATCTGTATCGTAGCAAACTGCCGGATATT

[0575] GAAATTAGCGATCATCTGCCGCTGCATAGCTATTGTTTTGAACGTGTTGCAGAATTTG

[0576] CAGATCGTCCGTGTCTGATTGATGGTGCAACCGATCGTACCTATTGCTTTAGCGAAGT

[0577] TGAACTGATTAGCCGTAAAGTTGCAGCAGGTCTGGCAAAACTGGGTCTGCAGCAGG

[0578] GTCAAGTTGTTATGCTGCTGCTGCCGAATTGTATTGAATTTGCCTTTGTTTTTATGGGT

[0579] GCAAGCGTTCGTGGTGCAATTGTTACCACCGCAAATCCGTTCTATAAACCGGGTGAA

[0580] ATTGCAAAACAGGCAAAAGCAGCCGGTGCACGTATTATTGTTACCCTGGCAGCATAT

[0581] GTTGAAAAACTGGCCGATCTGCAGAGCCATGATGTTCTGGTTATTACCATTGATGATG

[0582] CACCGAAAGAAGGTTGTCAGCATATTAGCGTTCTGACCGAAGCAGATGAAACCCAG

[0583] TGTCCGGCAGTGAAAATTCATCCTGATGATGTTGTTGCACTGCCGTATAGCAGCGGC

[0584] ACCACCGGTCTGCCGAAAGGTGTGATGCTGACCCATAAAGGTCTGGTTAGCAGCGT

[0585] TGCACAGCAGGTTGATGGTGAAAATCCGAATCTGTATTTTCATAGCGACGATGTTATT

[0586] CTGTGTGTGCTGCCGCTGTTTCATATCTATAGCCTGAATAGCGTGCTGCTGTGTGCAC

[0587] TGCGTGCCGGTGCAGCAACCCTGATTATGCAGAAATTTAACCTGACCACCTGTCTGG

[0588] AACTGATCCAGAAATACAAAGTTACCGTTGCACCGATTGTTCCGCCTATTGTGCTGG

[0589] ATATCACCAAAAGTCCGATTGTTAGCCAGTATGATGTGAGCAGCGTTCGCATTATTAT

[0590] GAGCGGTGCAGCTCCGCTGGGTAAAGAACTGGAAGATGCCCTGCGTGAACGTTTTC

[0591] CGAAAGCAATTTTTGGTCAAGGTTATGGTATGACAGAAGCAGGTCCGGTTCTGGCAA

[0592] TGAATCTGGCATTTGCCAAAAATCCGTTTCCGGTTAAAAGCGGTAGCTGTGGCACCG

[0593] TTGTTCGTAATGCACAGATTAAAATCCTGGATACCGAAACCGGTGAAAGCCTGCCGC

[0594] ATAACCAGGCAGGCGAAATTTGTATTCGTGGTCCGGAAATTATGAAGGGCTATATTAA

[0595] CGATCCGGAAAGCACCGCAGCAACAATTGATGAAGAAGGCTGGCTGCATACCGGTG

[0596] ATGTTGGTTATATCGATGATGATGAGGAAATCTTCATTGTCGATCGCGTGAAAGAGAT

[0597] CATCAAATATAAGGGTTTTCAGGTGGCACCAGCAGAGCTGGAAGCACTGCTGGTTG

[0598] CACATCCGAGCATTGCAGATGCAGCAGTTGTTCCGCAGAAACACGAAGAAGCAGGC

[0599] GAAGTTCCGGTTGCATTTGTTGTTAAAAGCAGCGAAATCAGCGAGCAAGAAATCAA

[0600] AGAATTTGTTGCCAAACAGGTGATCTTCTACAAAAAGATCCACCGCGTTTATTTTGTT

[0601] GATGCCATTCCGAAAAGCCCGAGCGGTAAAATTCTGCGTAAAGATCTGCGTAGCCGT

[0602] CTGGCAGCAAAATAA

[0603] Nucleotide sequence of cinnamic acid acyltransferase (At4CL2) (SEQ ID NO.17)

[0604] ATGACCACACAGGATGTTATTGTGAATGATCAGAACGATCAGAAACAGTGCAGCAA

[0605] CGATGTTATTTTTCGTAGCCGTCTGCCGGATATCTATATTCCGAATCATCTGCCGCTGC

[0606] ACGATTATATCTTTGAAAACATTAGCGAGTTTGCAGCCAAACCGTGTCTGATTAATGG

[0607] TCCGACCGGTGAAGTTTATACCTATGCAGATGTTCATGTTACCAGCCGTAAACTGGCA

[0608] GCAGGTCTGCATAATCTGGGTGTTAAACAGCATGATGTTGTGATGATTCTGCTGCCG

[0609] AATAGTCCGGAAGTTGTTCTGACCTTTCTGGCAGCCAGCTTTATTGGTGCAATTACCA

[0610] CCAGTGCAAATCCGTTTTTCACACCGGCAGAAATTAGCAAACAGGCAAAAGCAAGC

[0611] GCAGCAAAACTGATTGTTACCCAGAGCCGTTATGTGGACAAAATCAAAAATCTGCA

[0612] AAATGACGGTGTGCTGATTGTGACCACCGATAGTGATGCAATTCCGGAAAATTGTCT

[0613] GCGTTTTAGCGAACTGACCCAGAGCGAAGAACCGCGTGTTGATAGCATTCCTGAGA

[0614] AAATTTCACCGGAAGATGTTGTTGCACTGCCGTTTAGCAGCGGCACCACCGGTCTGC

[0615] CGAAAGGTGTTATGCTGACCCATAAAGGTCTGGTTACCAGCGTTGCACAGCAGGTTG

[0616] ATGGTGAAAATCCGAATCTGTATTTCAACCGTGATGATGTTATTCTGTGTGTGCTGCC

[0617] GATGTTTCATATTTATGCACTGAATAGCATTATGCTGTGTAGCCTGCGTGTTGGTGCAA

[0618] CCATTCTGATTATGCCGAAATTTGAAATTACCCTGCTGCTGGAACAAATTCAGCGTTG

[0619] TAAAGTTACCGTTGCAATGGTTGTTCCGCCTATTGTTCTGGCAATTGCAAAAAGTCC

[0620] GGAAACCGAAAAATATGATCTGAGCAGCGTTCGTATGGTTAAAAGCGGTGCAGCTC

[0621] CGCTGGGTAAAGAACTGGAAGATGCAATTAGCGCAAAATTTCCGAATGCCAAATTAG

[0622] GTCAAGGTTATGGTATGACCGAAGCAGGTCCGGTTCTGGCCATGAGCCTGGGTTTTG

[0623] CCAAAGAACCGTTTCCGGTTAAATCAGGTGCATGTGGCACCGTTGTTCGTAATGCAG

[0624] AAATGAAAATTCTGGACCCGGATACCGGTGATAGCCTGCCTCGTAATAAACCGGGTG

[0625] AAATCTGCATTCGTGGTAATCAGATTATGAAAGGCTATCTGAATGATCCGCTGGCAAC

[0626] CGCAAGCACCATTGATAAAGATGGTTGGCTGCATACAGGTGATGTGGGTTTTATCGAT

[0627] GATGATGACGAACTGTTTATTGTGGATCGTCTGAAAGAGCTGATCAAATACAAAGGT

[0628] TTTCAGGTTGCTCCTGCCGAACTGGAAAGCCTGCTGATTGGTCATCCGGAAATTAAT

[0629] GATGTTGCAGTTGTTGCCATGAAAGAAGAGGACGCAGGCGAAGTTCCGGTTGCATT

[0630] TGTTGTGCGTAGCAAAGATAGCAATATCAGCGAAGATGAGATCAAGCAGTTTGTGAG

[0631] TAAACAGGTGGTGTTCTATAAACGCATCAACAAAGTGTTTTTCACCGACAGCATTCC

[0632] GAAAGCACCGAGCGGTAAAATTCTGCGTAAAGACCTGCGTGCACGTCTGGCAAATG

[0633] GTCTGATGAATTAA

[0634] Nucleotide sequence of cinnamic acid acyltransferase (At4CL5) (SEQ ID NO.18)

[0635] ATGGTTCTGCAACAGCAGACCCATTTTCTGACCAAAAAGATCGATCAAGAAGATGA

[0636] AGAAGAAGAACCGAGCCACGATTTTATCTTTCGTAGCAAACTGCCGGATATCTTTATT

[0637] CCGAATCATCTGCCGCTGACCGATTATGTTTTTCAGCGTTTTAGCGGTGATGGTGATG

[0638] GCGATAGCAGCACCACCTGTATTATTGATGGTGCAACCGGTCGTATTCTGACCTATGC

[0639] AGATGTTCAGACCAATATGCGTCGTATTGCAGCAGGTATTCATCGTCTGGGTATTCGT

[0640] CATGGTGATGTTGTTATGCTGCTGCTGCCGAATAGTCCGGAATTTGCACTGAGCTTTC

[0641] TGGCAGTTGCATATCTGGGTGCAGTTAGCACCACCGCAAATCCGTTTTATACCCAGC

[0642] CGGAAATTGCAAAACAGGCAAAAGCAAGCGCAGCCAAAATGATTATTACCAAAAAG

[0643] TGCCTGGTGGACAAACTGACCAATCTGAAAAATGATGGTGTTCTGATTGTGTGCCTG

[0644] GATGATGATGGTGACAATGGTGTTGTTAGCAGCTCAGATGATGGTTGTGTTAGCTTTA

[0645] CCGAACTGACCCAGGCAGATGAAACCGAACTGCTGAAACCTAAAATTTCACCGGAA

[0646] GATACCGTTGCAATGCCGTATAGCAGCGGCACCACCGGTCTGCCGAAAGGTGTTATG

[0647] ATTACCCATAAAGGTCTGGTTACCAGCATTGCACAGAAAGTTGATGGTGAAAATCCG

[0648] AACCTGAATTTTACCGCCAATGATGTGATTCTGTGTTTTCTGCCGATGTTTCATATTTA

[0649] TGCACTGGATGCACTGATGCTGAGCGCAATGCGTACCGGTGCAGCACTGCTGATTGT

[0650] TCCGCGTTTTGAACTGAATCTGGTTATGGAACTGATCCAGCGTTATAAAGTTACCGTT

[0651] GTTCCGGTTGCGCCTCCGGTTGTTCTGGCATTTATCAAAAGTCCGGAAACGGAACGT

[0652] TATGATCTGAGCAGCGTTCGTATTATGCTGAGTGGTGCAGCAACCCTGAAGAAAGAA

[0653] CTGGAAGATGCAGTTCGTCTGAAATTTCCGAATGCAATTTTTGGCCAAGGTTATGGTA

[0654] TGACCGAAAGCGGCACCGTTGCCAAAAGCCTGGCATTTGCGAAAAACCCGTTTAAA

[0655] ACCAAAAGCGGTGCATGTGGCACCGTTATTCGTAATGCAGAAATGAAAGTTGTGGAT

[0656] ACCGAAACCGGTATTAGCCTGCCTCGTAATAAAAGTGGCGAAATTTGTGTTCGTGGT

[0657] CACCAGCTGATGAAAGGTTATCTGAATGATCCGGAAGCAACCGCACGTACCATTGAT

[0658] AAAGATGGTTGGCTGCATACCGGTGATATTGGTTTTGTTGATGATGACGACGAAATCT

[0659] TTATTGTGGATCGCCTGAAAGAGCTGATCAAATTCAAAGGCTATCAGGTTGCACCGG

[0660] CAGAGCTGGAAGCCCTGCTGATTAGCCATCCGAGCATTGATGATGCAGCAGTTGTTG

[0661] CAATGAAAGATGAAGTTGCCGATGAAGTTCCGGTGGCATTTGTTGCACGTAGCCAG

[0662] GGTAGCCAGCTGACCGAAGATGATGTTAAAAGCTATGTGAACAAACAGGTCGTGCA

[0663] CTACAAACGCATTAAGATGGTGTTTTTCATTGAGGTGATTCCGAAAGCAGTTAGCGG

[0664] CAAAATTCTGCGTAAAGACCTGCGTGCAAAACTGGAAACCATGTGTAGCAAATAA Nucleotide sequence of cinnamic acid acyltransferase (Os4CL) (SEQ ID NO.19)

[0665] ATGGGTAGCGTTGCAGCAGAAGAAGTTGTTGTTTTTCGTAGCAAACTGCCGGATATC

[0666] GAAATTGATAATAGCATGACCCTGCAAGAGTATTGTTTTGCACGTATGGCCGAAGTT

[0667] GGTGCACGTCCGTGTCTGATTGATGGTCAGACCGGTGAAAGCTATACCTATGCAGAA

[0668] GTTGAAAGCGCAAGCCGTCGTGCAGCAGCAGGTCTGCGTCGTATGGGTGTTGGTAA

[0669] AGGTGATGTTGTTATGAGCCTGCTGCGTAATTGTCCGGAATTTGCATTTAGCTTTCTG

[0670] GGTGCAGCACGTCTGGGTGCCGCAACCACCACCGCAAATCCGTTTTATACACCGCAT

[0671] GAAGTTCATCGTCAGGCCGAAGCAGCCGGTGCACGTGTTATTGTTACCGAAGCATGT

[0672] GCAGTTGAAAAAGTTCGTGAATTTGCAGCCGAACGTGGTGTTCCGGTTGTTACCGTT

[0673] GATGGTGCATTTGATGGTTGTGTTGAATTTCGTGAAGTTCTGGCAGCGGAAGAACTG

[0674] GATGCAGATGCCGATGTTCATCCTGATGATGTTGTGGCACTGCCGTATAGCAGCGGTA

[0675] CAACCGGTCTGCCGAAAGGTGTTATGCTGACCCATCGTAGCCTGATTACCAGCGTTG

[0676] CACAGCAGGTTGATGGCGAAAATCCGAATCTGTATTTCAGCAAAGATGATGTGATTC

[0677] TGTGTCTGCTGCCGCTGTTTCATATCTATAGCCTGAATAGCGTTCTGCTGGCTGGTCT

[0678] GCGTGCAGGTAGCACCATTGTTATTATGCGTAAATTTGATCTGGGTGCGCTGGTTGAT

[0679] CTGGTTCGTAAACATAACATTACCATTGCACCGTTTGTTCCGCCTATTGTTGTTGAAA

[0680] TTGCAAAAAGTCCGCGTGTTACCGCAGAGGATCTGGCAAGCATTCGTATGGTTATGA

[0681] GCGGTGCCGCACCGATGGGTAAAGATTTACAGGATGCATTTATGGCCAAAATTCCGA

[0682] ACGCAGTTTTAGGTCAAGGTTATGGTATGACCGAAGCAGGTCCGGTTCTGGCAATGT

[0683] GTCTGGCATTTGCAAAAGAACCGTTTAAAGTTAAAAGCGGTAGCTGTGGCACCGTT

[0684] GTTCGTAATGCAGAACTGAAAATTGTTGATCCGGATACCGGCACCAGCCTGGGTCGT

[0685] AATCAGAGTGGTGAAATTTGTATTCGTGGTGAGCAGATCATGAAAGGCTATCTGAAT

[0686] GATCCGGAAGCAACCAAAAATACATTGATGAAGATGGTTGGCTGCATACCGGTGAT

[0687] ATTGGTTTTGTGGATGATGATGACGAAATCTTTATTGTGGATCGCCTGAAAGAGATCA

[0688] TCAAATACAAAGGTTTTCAGGTTCCTCCGGCAGAACTGGAAGCACTGCTGATTACCC

[0689] ATCCGGAAATCAAAGATGCAGCAGTTGTTAGCATGAAAGATGATCTGGCAGGCGAA

[0690] GTTCCGGTGGCATTTATTGTTCGTACCGAAGGTAGCGAAATTACCGAAGATGAAATC

[0691] AAAAAGTTTGTGGCCAAAGAAGTGGTGTTCTACAAACGCATTAACAAGGTGTTTTT

[0692] CACCGATAGCATTCCGAAAAATCCGAGCGGTAAAATTCTGCGTAAAGACCTGCGTGC

[0693] CCGTCTGGCAGCAGGTATTCCGGATGCAGTTGCAGCCGCAGCAGCGGATGCACCGA

[0694] AAAGCAGCTAA

[0695] In the following examples and experimental examples, reagents and raw materials not specifically described are all commercially available products.

[0696] The present invention is directed to a non-naturally occurring recombinant microorganism that expresses genes encoding enzymes that catalyze the production of 1,2-pentanediol. The present invention has two pathways for producing 1,2-pentanediol: 1) 2-oxopentanoic acid is reduced to 2-hydroxypentanoic acid, which is then directly reduced to 2-hydroxypentanal and 1,2-pentanediol (e.g. Figure 1) and 2) 2-oxopentanoate is reduced to 2-hydroxypentanoate, which first forms 2-hydroxypentanoyl-CoA and then reduces to 2-hydroxypentanal and 1,2-pentanediol (e.g. Figure 2 ). Meanwhile, the synthesis of the intermediate product 2-oxopentanoic acid in the organism is based on two precursors: threonine and pyruvate.

[0697] The preferred embodiments of the present invention will be described in detail below in conjunction with the examples. It should be understood that the following examples are only for the purpose of illustration and are not intended to limit the scope of the present invention. Those skilled in the art may make various modifications and substitutions to the present invention without departing from the purpose and spirit of the present invention.

[0698] Example 1 Direct Reduction of 2-Oxopentanoic Acid to Form 1,2-Pentanediol

[0699] The conversion of 2-oxopentanoic acid to 1,2-pentanediol can be achieved through a three-step reaction: 2-oxopentanoic acid is converted to 2-hydroxypentanoic acid by 2-oxocarboxylic acid reductase, which is then reduced to 2-hydroxyvaleraldehyde by carboxylic acid reductase, and then reduced to 1,2-pentanediol by aldehyde reductase.

[0700] In this example, Escherichia coli was used as a host microorganism for strain transformation, and the nucleic acid encoding the above enzyme was expressed in Escherichia coli using existing molecular biological techniques. Specifically, lactate dehydrogenase ldh (SEQ ID NO.1, amino acid sequence BAN75276.1) of Lactobacillus casei was cloned into the pSTV28 vector to obtain the pSTV28-ldh plasmid ( Figure 6 At the same time, the gene encoding carboxylic acid reductase car (SEQ ID NO.2, amino acid sequence WP_005082584.1) and its activating factor gene sfp (EU882341.1), and the gene encoding aldehyde reductase yqhD (KEGG: JW2978) were cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain the plasmid pTrc99a-yqhD-sfp-car ( Figure 5 The two plasmids were then transformed into E. coli strain W3110 by electroporation to overexpress the enzymes involved in the conversion of 2-oxopentanoate to 1,2-pentanediol.

[0701] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the OD600 of the bacteria reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added at the same time. After continuing the fermentation for 48 hours, the 1,2-pentanediol product was detected by HPLC, and a yield of 698.8 mg / L was obtained.

[0702] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H 2 O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO 4 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8; Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0703] Example 2 Reduction of 2-oxopentanoic acid via acyl CoA to form 1,2-pentanediol

[0704] The conversion of 2-hydroxyvaleric acid to 1,2-pentanediol is achieved through a four-step reaction: 2-oxopentanoic acid is converted into 2-hydroxyvaleric acid by 2-oxocarboxylic acid reductase, the latter is catalyzed by CoA transferase to form 2-hydroxypentanoyl CoA, then reduced by acyl CoA reductase to form 2-hydroxyvaleraldehyde, and finally reduced by 2-hydroxyvaleraldehyde reductase to form 1,2-pentanediol.

[0705] In this example, Escherichia coli was used as a host microorganism for strain transformation, and the nucleic acid encoding the above enzymes was expressed in Escherichia coli using existing molecular biological techniques. The gene encoding CoA transferase fldA (SEQ ID NO.3, amino acid sequence WP_108429007.1), the gene encoding aldehyde reductase yqhD (KEGG: JW2978), and the gene encoding acyl CoA reductase maqu2507 (SEQ ID NO.4, amino acid sequence WP_011785966.1) were cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain the pTrc99a-yqhD-maqu2507-fldA plasmid ( Figure 8 ), which was co-transformed with plasmid pSTV28-ldh into Escherichia coli strain W3110 by electroporation to overexpress the enzymes involved in the conversion of 2-oxopentanoic acid to 1,2-pentanediol.

[0706] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added. After continuing the fermentation for 48 hours, 1,2-pentanediol product was detected by HPLC, and a yield of 299.2 mg / L was obtained (as shown in Figure 2A). Fig.13 ).

[0707] Example 3 Introducing cinnamic acylase (4CL) to improve the catalytic efficiency of fldA

[0708] Since the CoA donor of fldA is cinnamoyl-CoA, this example uses Escherichia coli as a host microorganism for strain modification, and uses existing molecular biology techniques to express multiple nucleic acids encoding cinnamic acylase in Escherichia coli. The catalytic efficiency of fldA is improved by screening and comparison.

[0709] Specifically, the yqhD-maqu2507 fragment was PCR amplified from pTrc99a-yqhD-maqu2507-fldA and cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain pTrc99a-yqhD-maqu2507. The cinnamic acylase Pc4CL (SEQ ID NO. 5, amino acid sequence P14912) gene was cloned into a Ptrc promoter by overlapping extension PCR with fldA, and then cloned into the linearized vector pTrc99a-yqhD-maqu2507 to obtain the pTrc99a-yqhD-maqu2507-Ptrc-fldA-Pc4CL plasmid ( Fig. 9 ). The plasmid was co-transformed into E. coli strain W3110 with plasmid pSTV28-ldh by electroporation to overexpress the enzymes related to the conversion of 2-oxopentanoate to 1,2-pentanediol.

[0710] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added. After continuing the fermentation for 48 hours, 1,2-pentanediol product was detected by HPLC, and a yield of 743.2 mg / L was obtained (as shown in Figure 2A). Fig.14 ).

[0711] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H2O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO 4 0.5, cinnamic acid 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8; Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0712] Example 4 Synthesis of 1,2-pentanediol from threonine

[0713] The conversion of threonine to 2-oxopentanoate can be achieved through multiple enzymatic steps (e.g. Figure 3Threonine is deaminated and dehydrated to generate 2-oxobutyrate, which is then converted to 2-oxopentanoate. 2-Oxopentanoate is converted to 2-hydroxypentanoate by 2-oxocarboxylic acid reductase, which is then catalyzed by CoA transferase to form 2-hydroxypentanoyl CoA, which is then reduced by acyl CoA reductase to form 2-hydroxypentanal, which is finally reduced by 2-hydroxypentanal reductase to form 1,2-pentanediol.

[0714] In this embodiment, Escherichia coli is used as a host microorganism for strain transformation, and the existing molecular biology technology is used to express the nucleic acid encoding the above enzyme in Escherichia coli. Specifically, the Escherichia coli gene cluster leuABCD (SEQ ID NO.6) is cloned into the pSTV28 vector to obtain pSTV28-leuABCD. At the same time, the coding genes of Escherichia coli threonine dehydratase ilvA (KEGG: JW3745) and Lactobacillus casei lactate dehydrogenase ldh (SEQ ID NO.1) are cloned into a Ptrc promoter by overlapping extension PCR, and then this fragment is inserted into the linearized plasmid pSTV28-leuABCD by one-step cloning to obtain the plasmid pSTV28-leuABCD-Ptrc-ilvA-ldh ( Fig.10 ), and this plasmid and plasmid pTrc99a-yqhD-maqu2507-Ptrc-fldA-Pc4CL were transformed into Escherichia coli strain W3110 by electroporation to overexpress the relevant enzymes from threonine to 1,2-pentanediol.

[0715] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the OD600 of the bacteria reached 0.8, 0.1 mM IPTG was added for induction, and 10 g / L threonine precursor was added at the same time. After continuing the fermentation for 48 hours, the 1,2-pentanediol product was detected by HPLC, and 2.3 g / L 1,2-pentanediol was obtained.

[0716] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H2O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO 4 0.5, cinnamic acid 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8; Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0717] Example 5 Synthesis of 1,2-pentanediol from pyruvic acid

[0718] The conversion of pyruvate to 2-oxopentanoate can be achieved through multiple enzymatic steps (e.g. Figure 4 Pyruvate is condensed with one molecule of acetyl-CoA to form 2-oxobutyrate, which is then converted to 2-oxopentanoate. 2-Oxopentanoate is converted to 2-hydroxypentanoate by 2-oxocarboxylic acid reductase, which is then catalyzed by CoA transferase to form 2-hydroxypentanoyl-CoA, which is then reduced by acyl-CoA reductase to form 2-hydroxypentanal, which is finally reduced by 2-hydroxypentanal reductase to form 1,2-pentanediol.

[0719] In this embodiment, Escherichia coli is used as a host microorganism for strain transformation, and the existing molecular biology technology is used to express the nucleic acid encoding the above enzyme in Escherichia coli. Specifically, the Escherichia coli gene cluster leuABCD is cloned into the pSTV28 vector to obtain pSTV28-leuABCD. At the same time, the coding genes of 2-methylmalate synthase cimA (SEQ ID NO.7, amino acid sequence WP_010870909.1) of Methanococcusjannaschii and lactate dehydrogenase ldh (SEQ ID NO.1) of Lactobacillus casei are cloned into a Ptrc promoter by overlapping extension PCR, and then this fragment is inserted into the linearized plasmid pSTV28-leuABCD by one-step cloning to obtain a plasmid.

[0720] pSTV28-leuABCD-Ptrc-cimA-ldh( Fig.11 ), the plasmid and the plasmid

[0721] pTrc99a-yqhD-maqu2507-Ptrc-fldA-Pc4CL was transformed into Escherichia coli strain W3110 by electroporation to overexpress the relevant enzymes from pyruvate to 1,2-pentanediol.

[0722] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the OD600 of the bacteria reached 0.8, 0.1 mM IPTG was added for induction. After continuing the fermentation for 48 hours, the 1,2-pentanediol product was detected by HPLC, and 671.3 mg / L 1,2-pentanediol was obtained.

[0723] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H 2 O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO4 0.5, cinnamic acid 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8; Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0724] Example 6 Increasing the production of 1,2-pentanediol by increasing the supply of NADPH

[0725] The pentose phosphate pathway was upregulated by overexpressing the Escherichia coli gene zwf, thereby increasing the NADPH supply. The coding gene of Escherichia coli glucose-6-phosphate 1-dehydrogenase zwf (KEGG: JW1841) was cloned into the linearized plasmid pSTV28-leuABCD-Ptrc-ilvA-ldh to obtain the plasmid

[0726] pSTV28-leuABCD-Ptrc-ilvA-ldh-zwf( Fig.12 ), the plasmid and the plasmid

[0727] pTrc99a-yqhD-maqu2507-Ptrc-fldA-Pc4CL was transformed into E. coli strain W3110 by electroporation to overexpress the enzymes involved in converting threonine to 1,2-pentanediol.

[0728] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 10 g / L threonine precursor was added. After continuing the fermentation for 48 hours, the 1,2-pentanediol product was detected by HPLC, and a 1,2-pentanediol yield of 3.2 g / L was obtained ( Fig.16 ).

[0729] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H2O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO 4 0.5, cinnamic acid 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8; Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0730] The technical solution of the present invention is further illustrated by experiments below.

[0731] Experimental Example 1 Screening experiment of acyl CoA transferase gene

[0732] The genes encoding the CoA transferase maqu2507 (WP_011785966.1), the gene encoding the aldehyde reductase yqhD (KEGG: JW2978), and five acyl-CoA transferases (CTs): fldA (SEQ ID NO.1), hadA (SEQ ID NO.8, amino acid sequence WP_095908939.1), pct (SEQ ID NO.9, amino acid sequence WP_014015705.1), Cat2 (SEQ ID NO.10, amino acid sequence WP_005873621.1), carB (SEQ ID NO.11, amino acid sequence WP_005873621.1), and dc (SEQ ID NO.12, amino acid sequence WP_005873622.1). NO.11, amino acid sequence WP_014355956.1), one of the five genes, was cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain five pTrc99a-yqhD-maqu2507-CT plasmids, which were co-transformed into the Escherichia coli strain W3110 with the plasmid pSTV28-ldh obtained in Example 1 by chemical transformation to overexpress the related enzymes from 2-oxopentanoic acid to 1,2-pentanediol.

[0733] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added. After continuing the fermentation for 48 hours, 1,2-pentanediol products were detected by HPLC. The highest yield strain carried the pTrc99a-yqhD-maqu2507-fldA plasmid and obtained a yield of 299.2 mg / L (as shown in Figure 2A). Fig.14 ).

[0734] Experimental Example 2 Screening experiment of acyl-CoA reductase gene

[0735] The gene encoding CoA transferase pct (SEQ ID NO.9), the gene encoding aldehyde reductase yqhD (KEGG: JW2978), and five acyl-CoA reductases (ACRs): maqu2507 (SEQ ID NO.4), maqu2220 (SEQ ID NO.12, amino acid sequence WP_011785687.1), bld (SEQ ID NO.13, amino acid sequence WP_077366605.1), ald (SEQ ID NO.14, amino acid sequence WP_077844109.1), pduP (SEQ ID NO.15, amino acid sequence WP_0778441109.1), and d-CoA reductase (SEQ ID NO.16, amino acid sequence WP_077844123.1) were cloned. NO.15, amino acid sequence WP_048265961.1), one of the five genes, was cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain five pTrc99a-yqhD-ACR-pct plasmids, which were transformed into the Escherichia coli strain W3110 by electroporation together with the plasmid pSTV28-ldh obtained in Example 1 to overexpress the related enzymes from 2-oxopentanoic acid to 1,2-pentanediol.

[0736] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added. After fermentation for 48 hours, HPLC was used to detect the 1,2-pentanediol product. The highest yield strain carried the pTrc99a-yqhD-maqu2507-pct plasmid ( Figure 7 ), and a yield of 153.4 mg / L was obtained (e.g. Fig.13 ).

[0737] Experimental Example 3 Screening experiment of cinnamic acylase gene

[0738] Since the CoA donor of fldA is cinnamoyl-CoA, this example uses Escherichia coli as a host microorganism for strain modification, and uses existing molecular biology techniques to express multiple nucleic acids encoding cinnamic acylase in Escherichia coli. The catalytic efficiency of fldA is improved by screening and comparison.

[0739] Specifically, the yqhD-maqu2507 fragment was PCR amplified from pTrc99a-yqhD-maqu2507-fldA and cloned into the pTrc99a vector under the control of the Ptrc promoter to obtain pTrc99a-yqhD-maqu2507. Five cinnamic acylase 4CL genes: Pt4CL (SEQ ID NO.16, amino acid sequence AAA92669.1), At4CL2 (SEQ ID NO.17, amino acid sequence NP_188761.1), At4CL5 (SEQ ID NO.18, amino acid sequence NP_188760), Os4CL (SEQ ID NO.19, amino acid sequence NP_001396278.1), and Pc4CL (SEQ ID NO.5) were cloned into a Ptrc promoter by overlapping extension PCR with fldA, and then cloned into the linearized vector pTrc99a-yqhD-maqu2507 to obtain five pTrc99a-yqhD-maqu2507-Ptrc-fldA-4CL plasmids. This group of plasmids were co-transformed into Escherichia coli strain W3110 with the plasmid pSTV28-ldh obtained in Example 1 by electroporation to overexpress the enzymes related to the conversion of 2-oxopentanoic acid to 1,2-pentanediol.

[0740] The obtained strain was cultured in a fermentation medium at a fermentation temperature of 37°C and a rotation speed of 200 rpm. When the bacterial OD600 reached 0.8, 0.1 mM IPTG was added for induction, and 1 g / L 2-oxopentanoic acid precursor was added. After continuing the fermentation for 48 hours, 1,2-pentanediol products were detected by HPLC. The highest yield strain carried the pTrc99a-yqhD-maqu2507-Ptrc-fldA-Pc4CL plasmid and obtained a yield of 743.2 mg / L (as shown in Figure 2A). Fig.15 ).

[0741] The fermentation medium formula is (g / L): glucose 20, Na 2 HPO 4 7H 2 O 12.8, KH 2 PO 4 3, NaCl 0.5, NH 4 Cl1,MgSO 4 0.5, cinnamic acid 0.5, yeast powder 5, CaCl 2 0.01, pH 6.8. Antibiotics (mg / L): ampicillin 100, chloramphenicol 35.

[0742] In summary, the present invention uses threonine as a raw material, and the enzymes involved include threonine dehydratase (ilvA), α-isopropylmalate synthase (leuA), α-isopropylmalate isomerase (leuC, D), β-isopropylmalate dehydrogenase (leuB), lactate dehydrogenase (ldh), acyl CoA transferase (fldA), cinnamic acylase (Pc4CL), acyl CoA reductase (maqu2507), aldehyde reductase (yqhD), and glucose-6-phosphate dehydrogenase (zwf). The recombinant microorganism obtained by using the above enzyme gene encoding plasmid can obtain 1,2-pentanediol with a yield of 3.2g / L through fermentation and culture. The present invention provides a biological method for synthesizing 1,2-pentanediol, and can significantly increase the yield of 1,2-pentanediol, which has broad application prospects.

Claims

1. A method for biosynthesizing 1,2-pentanediol, characterized in that: The method starts from 2-oxopentanoic acid, passes through 2-hydroxypentanoic acid and 2-hydroxypentanal intermediates, and synthesizes 1,2-pentanediol in three steps: Step 1: Reduction of 2-oxopentanoic acid to 2-hydroxypentanoic acid; Step 2: Reduction of 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde; Step 3: Reduction of 2-hydroxypentanal to 1,2-pentanediol.

2. The method according to claim 1, characterized in that Step 2 can obtain 2-hydroxyvaleraldehyde by a one-step reaction or a two-step reaction; the one-step reaction is: reducing 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde; the two-step reaction is: (1) converting 2-hydroxyvaleric acid to 2-hydroxyvaleryl-CoA; (2) reducing 2-hydroxyvaleryl-CoA to 2-hydroxyvaleraldehyde; The enzyme used to reduce 2-oxopentanoic acid to 2-hydroxypentanoic acid is 2-oxocarboxylic acid reductase; And / or, the enzyme used to convert 2-hydroxyvaleric acid into 2-hydroxyvaleryl-CoA is acyl CoA transferase or acyl CoA ligase; and / or, the enzyme used to reduce 2-hydroxyvaleryl-CoA to 2-hydroxyvaleraldehyde is acyl-CoA reductase; And / or, the enzyme used to reduce 2-hydroxyvaleric acid to 2-hydroxyvaleraldehyde is carboxylic acid reductase; And / or, the enzyme used to reduce 2-hydroxyvaleraldehyde to 1,2-pentanediol is aldehyde reductase; And / or, cinnamate acylase is overexpressed in the step of converting 2-hydroxyvalerate into 2-hydroxyvaleryl-CoA.

3. The method according to claim 2, characterized in that The method is to reduce and transform under at least one of the conditions of crude enzyme solution, a combination of purified enzymes, immobilized enzymes and whole cells.

4. The method according to claim 3, characterized in that The whole cell is a recombinant microorganism; the starting strain of the recombinant microorganism is selected from at least one of Escherichia coli, Corynebacterium glutamicum, Bacillus subtilis, Saccharomyces cerevisiae and Yarrowia lipolytica.

5. The method according to claim 2, characterized in that The 2-oxocarboxylate reductase is selected from the following gene encodings: one or more of ldh, ldhA, panE, ara, ara1, ara2, araDH, YGL039W, adh, and CXQ87_003259; And / or, the acyl-CoA transferase or acyl-CoA ligase is selected from one or more of the following genes encoding: fldA, hadA, pct, Cat2, carB, 4CL, PaaK, Cat1, PGN_0725, PGN_1341, PGN_1888, fadD; And / or, the acyl-CoA reductase is selected from one or more of the following genes encoding: maqu2220, maqu2507, bld, ald, pdcD, pduP, adhE, FAR, acr, hom, fadB, fadJ, hbd, phaB, phbB, mdh, ldh; And / or, the carboxylate reductase is encoded by a CAR gene; And / or, the aldehyde reductase is selected from the following gene encoding: one or more of yqhD, ADH2, adh, bdh, yahK, adhA, 4-hbd, dhaT, mmsb, P84067, adhI, alrA, sucD, and bld; And / or, the cinnamate acylase is selected from one or more of the following gene encodings: Pt4CL, At4CL2, At4CL5, Os4CL, Pc4CL.

6. The method according to any one of claims 1 to 5, characterized in that: The method also includes the step of increasing the yield of 1,2-pentanediol by adding precursors, cofactors or increasing the concentrations of precursors and cofactors based on the above method; the precursor is at least one of threonine and pyruvate, and the cofactor is NAD(P)H, NAD(P) + , ATP or at least one of them.

7. The method according to any one of claims 1 to 6, characterized in that: Taking the recombinant microorganism that has expressed the ldh, CAR, and yqhD genes, culturing them in a culture medium, inducing the expression of the recombinant microorganism, adding 2-oxopentanoic acid, and continuing the fermentation to obtain 1,2-pentanediol; Alternatively, a recombinant microorganism expressing the ldh, fldA, maqu2507, and yqhD genes is taken, cultured in a culture medium, the expression of the recombinant microorganism is induced, 2-oxopentanoic acid is added, and fermentation is continued to obtain 1,2-pentanediol; Alternatively, a recombinant microorganism expressing ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, 2-oxopentanoic acid is added, and fermentation is continued to obtain 1,2-pentanediol; Alternatively, a recombinant microorganism expressing ilvA, leuABCD, ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, threonine is added, and fermentation is continued to obtain 1,2-pentanediol; Alternatively, a recombinant microorganism expressing cimA, leuABCD, ldh, fldA, maqu2507, yqhD, and Pc4CL genes is cultured in a medium containing cinnamic acid to induce expression of the recombinant microorganism, pyruvic acid is added, and fermentation is continued to obtain 1,2-pentanediol; Alternatively, a recombinant microorganism that overexpresses ilvA, leuABCD, ldh, fldA, maqu2507, yqhD, Pc4CL, and zwf genes is cultured in a culture medium including cinnamic acid to induce expression of the recombinant microorganism, threonine is added, and fermentation is continued to obtain 1,2-pentanediol.

8. 1,2-Pentanediol is prepared according to the method according to any one of claims 1 to 7.

9. A recombinant microorganism, characterized in that The recombinant microorganism overexpresses genes for 2-oxocarboxylic acid reductase, carboxylic acid reductase, acyl-CoA reductase, acyl-CoA transferase, acyl-CoA ligase, cinnamate acylase and / or aldehyde reductase; Preferably, the starting strain of the recombinant microorganism is selected from one or more of Escherichia coli, Corynebacterium glutamicum, Bacillus subtilis, Saccharomyces cerevisiae and Yarrowia lipolytica.

10. The recombinant microorganism according to claim 9, characterized in that The 2-oxocarboxylate reductase is selected from the following gene encodings: one or more of ldh, ldhA, panE, ara, ara1, ara2, araDH, YGL039W, adh, and CXQ87_003259; And / or, the acyl-CoA transferase or acyl-CoA ligase is selected from one or more of the following genes encoding: fldA, hadA, pct, Cat2, carB, 4CL, PaaK, Cat1, PGN_0725, PGN_1341, PGN_1888, fadD; And / or, the acyl-CoA reductase is selected from one or more of the following genes encoding: maqu2220, maqu2507, bld, ald, pdcD, pduP, adhE, FAR, acr, hom, fadB, fadJ, hbd, phaB, phbB, mdh, ldh; And / or, the carboxylate reductase is encoded by a CAR gene; And / or, the aldehyde reductase is selected from the following gene encoding: one or more of yqhD, ADH2, adh, bdh, yahK, adhA, 4-hbd, dhaT, mmsb, P84067, adhI, alrA, sucD, and bld; And / or, the cinnamate acylase is selected from one or more of the following gene encodings: Pt4CL, At4CL2, At4CL5, Os4CL, Pc4CL.

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