KR20210038805A - Methylotroph with enhanced 1,2-propylene glycol productivity and method for producing 1,2-propylene glycol using the same - Google Patents

Abstract

The present invention relates to methylotrophic bacteria with enhanced 1,2-propylene glycol (propanediol; PDO) productivity and to a 1,2-PDO production method using the same. As a result of constructing a 1,2-propylene glycol (propanediol; PDO) biosynthetic pathway by introducing a foreign gene into Methylobacterium extorquens, the 1,2-PDO is produced, thereby effectively used in the production of 1,2-PDO.

Description

Methylotroph with enhanced 1,2-propylene glycol productivity and method for producing 1,2-propylene glycol using the methylotroph with enhanced 1,2-propylene glycol productivity the same}

The present invention relates to a methyl nutrient bacterium with enhanced productivity of 1,2-propylene glycol (PDO) and a method for producing 1,2-PDO using the same, and in more detail, 1,2-propylene glycol by itself The present invention relates to a method for producing 1,2-propylene glycol using methanol as a carbon source by establishing a biosynthetic pathway in methyl nutrient bacteria that cannot produce Cole.

1,2-Propanediol (PDO) is a building block in various fields such as antifreeze, polyester resin, pharmaceuticals, cosmetics, detergent, etc. It is the material used. Due to this high availability, over 1 trillion lbs per year are sold in the US alone, and demand is estimated at over 3 trillion lbs/yr worldwide.

Meanwhile, most 1,2-PDOs are produced through petrochemical processes. Propylene, a product of steam cracking, is converted to propylene oxide, which is hydrolyzed to produce 1,2-PDO, and harmful intermediates and by-products are produced. In order to replace this process with a more sustainable and eco-friendly bio process, attempts have been made to produce 1,2-PDO using various microorganisms.

Representative examples include Escherichia coli , Corynebacterium glutamicum , Saccharomyces cerevisiae , Clostridium thermosaccharolyticum , and most of them are glucose. ), rhamnose, fructose, such as sugar or glycerol was used as a substrate.

Due to the fact that the price of sugar continues to rise and it causes food problems, research and demand for raw materials that can replace sugar are actively being conducted. Methanol has been attracting attention from the past as a material that can be synthesized from shale gas or biogas whose main component is methane, and it is economical at a lower price than sugar, and its solubility in aqueous solution is close to infinity.

In addition, since methanol is toxic, contamination by other microorganisms that may occur during the fermentation process can be reduced, and since it is supplied to a mineral medium and used as a substrate, cost in the downstreaming process can be reduced. Therefore, an attempt was made to develop a strain that produces 1,2-PDO using methylotroph, which can use methanol as a carbon source.

Accordingly, the present inventors confirmed that 1,2-PDO was produced as a result of constructing a 1,2-propanediol (PDO) biosynthetic pathway by introducing a foreign gene into Methylobacterium extorquens. I did.

Accordingly, an object of the present invention is to provide a recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase. .

Another object of the present invention is to provide a methylotroph containing a nucleic acid sequence encoding Lactoylglutathione lyase in an inactivated state.

Another object of the present invention is to provide a composition for producing 1,2-propylene glycol (propanediol; PDO) comprising methyltrophic bacteria comprising a nucleic acid sequence encoding lactoyl glutathione lyase in an inactivated state will be.

Another object of the present invention is to provide a method for producing 1,2-PDO comprising a culturing step of culturing a methyltrophic bacterium containing a nucleic acid sequence encoding lactoyl glutathione lyase in an inactivated state.

Another object of the present invention relates to the use of a methyltrophic bacteria for the production of 1,2-PDO comprising a nucleic acid sequence encoding lactoyl glutathione lyase in an inactivated state.

The present invention relates to a composition for producing 1,2-propylene glycol (PDO) using methyl nutrient bacteria and a production method thereof, and the method according to the present invention produces 1,2-PDO using methanol as a carbon source. Show how to do it.

The present inventors confirmed that 1,2-PDO was produced as a result of constructing a 1,2-PDO biosynthetic pathway by introducing a foreign gene into Methylobacterium extorquens.

Hereinafter, the present invention will be described in more detail.

One aspect of the present invention is a recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase.

The term "formolease" herein refers to an enzyme that converts formaldehyde to dihydroxyacetone (DHA), and may be encoded by the fls gene consisting of a nucleic acid sequence represented by SEQ ID NO: 11, but this It is not limited.

The term "dihydroxyacetone kinase" as used herein refers to an enzyme that converts dihydroxyacetone (DHA) to dihydroxyacetone phosphate, and consists of a nucleic acid sequence represented by SEQ ID NO: 12. It may be encoded by the dak1 gene derived from Saccharomyces cerevisiae, but is not limited thereto.

The recombinant vector includes a nucleic acid sequence encoding methylglyoxal synthase, a nucleic acid sequence encoding methylglyoxal reductase or lactaldehyde reductase, and glycerol dehydrogenase. ) May additionally include one or more selected from the group consisting of a nucleic acid sequence encoding.

The term "methylglyoxal synthase" herein refers to an enzyme that converts dihydroxyacetone phosphate (DHAP) to methylglyoxal (MG), and consists of a nucleic acid sequence represented by SEQ ID NO: 2 It may be encoded by the mgsA gene, but is not limited thereto.

The term "glycerol dehydrogenase" as used herein refers to an enzyme that converts hydroxyacetone into 1,2-PDO, and may be encoded by a gldA gene consisting of a nucleic acid sequence represented by SEQ ID NO: 4, but is limited thereto. It is not.

The term "vector" means a means for expressing a gene of interest in a host cell. For example, plasmid vectors, cosmid vectors and bacteriophage vectors, adenovirus vectors, retroviral vectors, and viral vectors such as adeno-associated virus vectors. Vectors that can be used as the recombinant vector include plasmids often used in the art (e.g., pSC101, pGV1106, pACYC177, ColE1, pKT230, pME290, pBR322, pUC8/9, pUC6, pBD9, pHC79, pIJ61, pLAFR1, pHV14 , pGEX series, pET series, and pUC19, etc.), phage (eg, λgt4λ B, λ-Charon, λβ, and M13, etc.) or viruses (eg, SV40, etc.), but are not limited thereto. .

The recombinant vector can typically be constructed as a vector for cloning or as a vector for expression. The expression vector may be a conventional one used in the art to express foreign proteins in plants, animals, or microorganisms. The recombinant vector can be constructed through various methods known in the art.

The recombinant vector can be constructed using a prokaryotic cell or a eukaryotic cell as a host. For example, when the vector used is an expression vector and a prokaryotic cell is used as a host, a strong promoter capable of promoting transcription (e.g., pLλ promoter, CMV promoter, trp promoter, lac promoter, tac promoter, T7 Promoter, etc.), a ribosome binding site for initiation of translation, and a transcription/translation termination sequence are generally included. In the case of eukaryotic cells as a host, the origin of replication operating in eukaryotic cells included in the vector includes the f1 origin of replication, SV40 origin of replication, pMB1 origin of replication, adeno origin of replication, AAV origin of replication, BBV origin of replication, etc. It is not limited. In addition, a promoter derived from the genome of a mammalian cell (e.g., metallothionine promoter) or a promoter derived from mammalian virus (e.g., adenovirus late promoter, vaccinia virus 7.5K promoter, SV40 promoter, The cytomegalovirus promoter and the tk promoter of HSV) can be used and generally have a polyadenylation sequence as the transcription termination sequence.

In an example of the present invention, a transformant may be produced by inserting a recombinant vector into a host cell, and the transformant may be obtained by introducing the recombinant vector into an appropriate host cell.

The host cell is a cell capable of stably and continuously cloning or expressing the expression vector, and any host cell known in the art may be used.

Host cells used in the present invention may include E. coli, yeast, animal cells, plant cells, insect cells, and the like, and prokaryotic cells include, for example, E. coli JM109, E. coli BL21, E. strains of the genus Bacillus such as coli RR1, E. coli LE392, E. coli B, E. coli X 1776, E. coli W3110, Bacillus subtilis, Bacillus thuringensis, and Salmonella typhimurium, Serratia marsessons, and There are enterobacteriaceae strains such as various Pseudomonas species, and when transforming into eukaryotic cells, as host cells, yeast (Saccharomyce cerevisiae), insect cells, plant cells and animal cells such as Sp2/0, CHO (Chinese hamster ovary) K1, CHO DG44, PER.C6, W138, BHK, COS7, 293, HepG2, Huh7, 3T3, RIN, MDCK cell lines, etc. may be used, but are not limited thereto.

Transport (introduction) of the polynucleotide or a recombinant vector containing the same into a host cell may use a transport method well known in the art. For example, when the host cell is a prokaryotic cell, a CaCl2 method or an electroporation method can be used, and when the host cell is a eukaryotic cell, a microinjection method, calcium phosphate precipitation method, electroporation method, liposome Mediated transfection methods and gene bombardment may be used, but are not limited thereto.

The method of selecting the transformed host cell can be easily carried out according to a method well known in the art using a phenotype expressed by a selection label. For example, when the selection marker is a specific antibiotic resistance gene, the transformant can be easily selected by culturing the transformant in a medium containing the antibiotic.

Another aspect of the present invention is a methylotroph containing a nucleic acid sequence encoding a lactoylglutathione lyase in an inactivated state.

The nucleic acid sequence may be one or more selected from the group consisting of a nucleic acid sequence represented by SEQ ID NO: 17 and a nucleic acid sequence represented by SEQ ID NO: 18, but is not limited thereto.

The methylotrophic bacteria may be transformed with a recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding formolease and a nucleic acid sequence encoding dihydroxyacetone kinase.

The recombinant vector contains at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a methylglyoxal reductase or a lactaldehyde reductase, and a nucleic acid sequence encoding a glycerol dehydrogenase. It may be additionally included, but is not limited thereto.

The methylotrophic bacteria are at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a nucleic acid sequence encoding a methylglyoxal reductase or lactaldehyde reductase. It may be transformed with a recombinant vector containing.

The methylotrophic bacteria are Methylobacterium extorquens , Methylobacterium suomiense , Methylobacterium platani, Methylobacterium adhaesivum ), methylobacterium soli (Methylobacterium soli) and methylobacterium chloromethanicum (Methylobacterium chloromethanicum ) may be one or more selected from the group consisting of, for example, methylobacterium extorkens. However, it is not limited thereto.

Another aspect of the present invention is a composition for the production of 1,2-propylene glycol (PDO) comprising methyltrophic bacteria comprising a nucleic acid sequence encoding a lactoyl glutathione degrading enzyme in an inactivated state.

The methylotrophic bacteria may be transformed with a recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding formolease and a nucleic acid sequence encoding dihydroxyacetone kinase.

The recombinant vector contains at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a methylglyoxal reductase or a lactaldehyde reductase, and a nucleic acid sequence encoding a glycerol dehydrogenase. It may be additionally included, but is not limited thereto.

The methylotrophic bacteria are at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a nucleic acid sequence encoding a methylglyoxal reductase or lactaldehyde reductase. It may be transformed with a recombinant vector containing.

The methylotrophic bacteria are Methylobacterium extorkens, Methylobacterium suomiens, Methylobacterium platinum, Methylobacterium ad hashboom, Methylobacterium solai and Methylobacterium chloromethanicum. It may be one or more selected from the group consisting of, for example, may be methylobacterium extorcense, but is not limited thereto.

Another aspect of the present invention is a method for producing 1,2-PDO comprising a culturing step of culturing a methyl nutrient bacterium containing a nucleic acid sequence encoding a lactoyl glutathione degrading enzyme in an inactivated state.

The methylotrophic bacteria may be transformed with a recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding formolease and a nucleic acid sequence encoding dihydroxyacetone kinase.

The recombinant vector contains at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a methylglyoxal reductase or a lactaldehyde reductase, and a nucleic acid sequence encoding a glycerol dehydrogenase. It may be additionally included, but is not limited thereto.

The methylotrophic bacteria are at least one selected from the group consisting of a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a nucleic acid sequence encoding a methylglyoxal reductase or lactaldehyde reductase. It may be transformed with a recombinant vector containing.

The methylotrophic bacteria are Methylobacterium extorkens, Methylobacterium suomiens, Methylobacterium platinum, Methylobacterium ad hashboom, Methylobacterium solai and Methylobacterium chloromethanicum. It may be one or more selected from the group consisting of, for example, may be methylobacterium extorcense, but is not limited thereto.

The present invention relates to a methylotrophic bacteria with enhanced productivity of 1,2-propylene glycol (PDO) and a method for producing 1,2-PDO using the same, in the methylobacterium extorquens (Methylobacterium extorquens) As a result of constructing a 1,2-propanediol (PDO) biosynthetic pathway by introducing a foreign gene, 1,2-PDO is produced, which can be effectively used for the production of 1,2-PDO.

FIG. 1 shows methylglyoxal synthase (mgsA) and glycerol dehydrogenase (gldA), and methylglyoxal reductase or lactaldehyde reductase (fucO). It is a schematic diagram of the 1,2-propylene glycol (propanediol; PDO) biosynthetic pathway constructed by doing so.
2 is a schematic diagram of a 1,2-PDO biosynthetic pathway according to an embodiment of the present invention.
3 is a graph showing the culture results of M. extorquens AM1::pMEV:: mgsA :: yqhD :: gldA strain (MPG001) according to an embodiment of the present invention.
4 is a graph showing the culture results of M. extorquens AM1::pMEV:: mgsA :: yqhD :: gldA :: fls :: dak1 strain (MPG011) according to an embodiment of the present invention.
5 is a graph showing the culture results of M. extorquens AM1gloA::pMEV:: mgsA :: yqhD :: gldA strain (MPG101) according to an embodiment of the present invention.
6 is a graph showing the culture results of M. extorquens AM1gloA::pMEV:: mgsA :: yqhD :: gldA :: fls :: dak1 strain (MPG111) according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited by these examples.

Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight)% for solids/solids, (weight/volume)% for solids/liquids, and Liquid/liquid is (vol/vol)%.

Example 1: Construction of 1,2-propylene glycol biosynthetic pathway

Methylobacterium extorquens (Methylobacterium extorquens) AM1, which can use methanol as a carbon source, has a known metabolism, and has developed a genetic manipulation tool, was used as a recombinant host. M. extorquens AM1 wild type was unable to produce 1,2-PDO, so a foreign gene was introduced to construct a 1,2-PDO biosynthetic pathway as shown in FIG. 1. The blue arrow indicates the route taken.

There are two types of 1,2-PDO production pathways, one through hydroxyacetone and one through L-lactaldehyde. A recombinant strain having a pathway through hydroxyacetone or L-lacaldehyde was constructed.

1-1. Preparation of a recombinant strain having a hydroxyacetone pathway

The pathway through hydroxyacetone is methylglyoxal synthase, an enzyme that converts dihydroxyacetone phosphate (DHAP) derived from Bacillus subtilis into methylglyoxal (MG). MgsA gene encoding E. coli , yqhD gene encoding methylglyoxal reductase, an enzyme that converts MG derived from Escherichia coli to hydroxyacetone, and hydroxyacetone derived from E. coli as 1,2-PDO The gldA gene encoding the converting enzyme, glycerol dehydrogenase, was introduced into the pMEV vector and overexpressed using this vector. The two strains developed are M. extorquens AM1::pMEV:: mgsA :: yqhD :: gldA .

The sequence information of the pMEV vector is the same as the sequence of SEQ ID NO: 1.

The sequence information of related genes is shown in Table 1 below.

order
number designation order 2 mgsA coding nucleic acid sequence ATGAAAATTGCTTTGATCGCGCATGACAAGAAAAAACAGGATATGGTTCAATTTACGACTGCCTATCGGGATATTTTAAAGAATCATGATCTATACGCAACCGGAACCACAGGGTTGAAAATTCATGAGGCGACAGGTCTTCAAATTGAACGTTTTCAATCCGGCCCTTTAGGGGGAGACCAGCAAATCGGTGCACTGATCGCTGCCAATGCACTCGATCTTGTCATTTTTTTGCGCGACCCGCTGACCGCGCAGCCGCATGAACCGGATGTCTCGGCATTAATCCGTTTATGTGATGTGTATTCCATTCCGCTCGCCACAAATATGGGTACTGCGGAAATTCTTGTGCGCACACTTGATGAAGGTGTTTTCGAATTCCGTGACCTTCTTCGGGGAGAAGAGCCGAATGTATAA 3 yqhD coding nucleic acid sequence ATGAACAACTTTAATCTGCACACCCCAACCCGCATTCTGTTTGGTAAAGGCGCAATCGCTGGTTTACGCGAACAAATTCCTCACGATGCTCGCGTATTGATTACCTACGGCGGCGGCAGCGTGAAAAAAACCGGCGTTCTCGATCAAGTTCTGGATGCCCTGAAAGGCATGGACGTGCTGGAATTTGGCGGTATTGAGCCAAACCCGGCTTATGAAACGCTGATGAACGCCGTGAAACTGGTTCGCGAACAGAAAGTGACTTTCCTGCTGGCGGTTGGCGGCGGTTCTGTACTGGACGGCACCAAATTTATCGCCGCAGCGGCTAACTATCCGGAAAATATCGATCCGTGGCACATTCTGCAAACGGGCGGTAAAGAGATTAAAAGCGCCATCCCGATGGGCTGTGTGCTGACGCTGCCAGCAACCGGTTCAGAATCCAACGCAGGCGCGGTGATCTCCCGTAAAACCACAGGCGACAAGCAGGCGTTCCATTCTGCCCATGTTCAGCCGGTATTTGCCGTGCTCGATCCGGTTTATACCTACACCCTGCCGCCGCGTCAGGTGGCTAACGGCGTAGTGGACGCCTTTGTACACACCGTGGAACAGTATGTTACCAAACCGGTTGATGCCAAAATTCAGGACCGTTTCGCAGAAGGCATTTTGCTGACGCTAATCGAAGATGGTCCGAAAGCCCTGAAAGAGCCAGAAAACTACGATGTGCGCGCCAACGTCATGTGGGCGGCGACTCAGGCGCTGAACGGTTTGATTGGCGCTGGCGTACCGCAGGACTGGGCAACGCATATGCTGGGCCACGAACTGACTGCGATGCACGGTCTGGATCACGCGCAAACACTGGCTATCGTCCTGCCTGCACTGTGGAATGAAAAACGCGATACCAAGCGCGCTAAGCTGCTGCAATATGCTGAACGCGTCTGGAACATCACTGAAGGTTCCGATGATGAGCGTATTGACGCCGCGATTGCCGCAACCCGCAATTTCT TTGAGCAATTAGGCGTGCCGACCCACCTCTCCGACTACGGTCTGGACGGCAGCTCCATCCCGGCTTTGCTGAAAAAACTGGAAGAGCACGGCATGACCCAACTGGGCGAAAATCATGACATTACGTTGGATGTCAGCCGCCGTATATACGAAGCCGCCCGCTAA 4 gldA coding nucleic acid sequence ATGGACCGCATTATTCAATCACCGGGTAAATACATCCAGGGCGCTGATGTGATTAATCGTCTGGGCGAATACCTGAAGCCGCTGGCAGAACGCTGGTTAGTGGTGGGTGACAAATTTGTTTTAGGTTTTGCTCAATCCACTGTCGAGAAAAGCTTTAAAGATGCTGGACTGGTAGTAGAAATTGCGCCGTTTGGCGGTGAATGTTCGCAAAATGAGATCGACCGTCTGCGTGGCATCGCGGAGACTGCGCAGTGTGGCGCAATTCTCGGTATCGGTGGCGGAAAAACCCTCGATACTGCCAAAGCACTGGCACATTTCATGGGTGTTCCGGTAGCGATCGCACCGACTATCGCCTCTACCGATGCACCGTGCAGCGCATTGTCTGTTATCTACACCGATGAGGGTGAGTTTGACCGCTATCTGCTGTTGCCAAATAACCCGAATATGGTCATTGTCGACACCAAAATCGTCGCTGGCGCACCTGCACGTCTGTTAGCGGCGGGTATCGGCGATGCGCTGGCAACCTGGTTTGAAGCGCGTGCCTGCTCTCGTAGCGGCGCGACCACCATGGCGGGCGGCAAGTGCACCCAGGCTGCGCTGGCACTGGCTGAACTGTGCTACAACACCCTGCTGGAAGAAGGCGAAAAAGCGATGCTTGCTGCCGAACAGCATGTAGTGACTCCGGCGCTGGAGCGCGTGATTGAAGCGAACACCTATTTGAGCGGTGTTGGTTTTGAAAGTGGTGGTCTGGCTGCGGCGCACGCAGTGCATAACGGCCTGACCGCTATCCCGGACGCGCATCACTATTATCACGGTGAAAAAGTGGCATTCGGTACGCTGACGCAGCTGGTTCTGGAAAATGCGCCGGTGGAGGAAATCGAAACCGTAGCTGCCCTTAGCCATGCGGTAGGTTTGCCAATAACTCTCGCTCAACTGGATATTAAAGAAGATGTCCCGGCGAAAATGCGAATTGTGGCAGAAGCGGCATGTGCAGAAGGTG AAACCATTCACAACATGCCTGGCGGCGCGACGCCAGATCAGGTTTACGCCGCTCTGCTGGTAGCCGACCAGTACGGTCAGCGTTTCCTGCAAGAGTGGGAATAA

The strain was produced by multi-insert cloning using an infusion enzyme, and EcoR1 and Sac1 were used as restriction enzymes. Primers used for cloning of the M. extorquens AM1::pMEV::mgsA::yqhD::gldA strain (MPG001) are SEQ ID NOs: 5 to 10, as shown in Table 2 below. The underlined sequence is the ribosome binding site (RBS).

order
number designation order 5 Forward Primer 1 GAGAGACAGCGAATTC CCGCGAGGGAATTCCCAAAGGCCAGGTATTTT ATGAAAATTGCTTTGATCGC 6 Reverse primer 1 TGGGTCTATTGGGAGTTATACATTCGGCTCTTCTCCC 7 Forward primer 2 CTCCCAATAGACCCAAAGGCGGTAGAGAA ATGAACAACTTTAATCTGCACACCC 8 Reverse primer 2 TTGCGCTTAGTTGCCTTAGCGGGCGGCTTCGTATATA 9 Forward Primer 3 GGCAACTAAGCGCAAAGATCGAGGTAATAA ATGGACCGCATTATTCAATCA 10 Reverse primer 3 CGGGGCTGGCTTAAGAGCTCTTATTCCCACTCTTGCAGGAA

1-2. Preparation of recombinant strains with increased DHAP flow

DHAP is a metabolite of M. extorquens AM1 and a precursor of the previously inserted 1,2-PDO biosynthetic pathway, and is converted to methylglyoxal by methylglyoxal (MG) biosynthesis. M. extorquens To convert methanol, the carbon source of AM1, to DHAP, a complicated step is required.

Accordingly, a more direct DHAP synthesis route was introduced to enhance the DHAP flux. Since M. extorquens AM1 has a rich formaldehyde flow, Formolase, an enzyme that converts formaldehyde to dihydroxyacetone (DHA), and DHA, to dihydroxyacetone phosphate. The enzyme dihydroxyacetone kinase was overexpressed.

Although papers have been reported in terms of enabling a new carbon assimilation pathway, formolase has not been reported for the purpose of producing such a specific substance, and there has been no precedent of operating formolase in strains other than E. coli. Is estimated.

The gene fls encoding formolase was codon-optimized for AM1. There are various genes encoding dihydroxyacetone kinase, and in this study, dak1 derived from Saccharomyces cerevisiae was used. A plasmid pMEV build on existing :: mgsA :: yqhD :: gldA new plasmids by introducing additional fls and dak1 the pMEV :: mgsA :: yqhD :: gldA :: fls :: dak1 was built with the plasmid M extorquens AM1::pMEV:: mgsA :: yqhD :: gldA :: fls :: dak1 strain (MPG011) was developed.

Related gene sequences are shown in Table 3 below.

order
number designation order 11 Codon optimized fls coding
Nucleic acid sequence ATGGCCATGATCACGGGCGGCGAGCTGGTCGTGCGCACGCTGATCAAGGCGGGTGTCGAGCACCTCTTTGGTCTCCACGGTATCCACATCGACACCATCTTCCAGGCCTGCCTGGACCATGATGTCCCGATCATCGACACCCGCCACGAGGCGGCAGCCGGACATGCGGCGGAAGGCTACGCCCGCGCCGGCGCCAAGCTCGGCGTCGCGCTTGTGACGGCGGGCGGCGGCTTCACCAACGCGGTGACCCCGATCGCCAATGCCCGCACCGACCGCACGCCGGTCCTCTTCCTTACCGGCTCGGGCGCCCTCAGGGATGACGAGACGAACACCCTCCAGGCGGGGATCGACCAGGTCGCGATGGCCGCCCCTATCACCAAGTGGGCCCATCGGGTAATGGCCACCGAACACATCCCTCGGCTGGTCATGCAGGCGATTCGCGCCGCGCTGAGCGCCCCCCGCGGCCCAGTTCTGCTCGACCTCCCGTGGGACATCCTCATGAACCAGATTGACGAAGATAGCGTCATCATCCCGGACCTCGTGCTGTCTGCCCACGGCGCGCACCCCGATCCCGCGGACCTCGATCAGGCCCTGGCGCTGCTCCGCAAGGCCGAGCGTCCGGTTATCGTTCTGGGCAGTGAGGCCAGCCGCACGGCCCGAAAGACGGCCCTCTCGGCCTTCGTCGCGGCCACGGGCGTGCCGGTCTTCGCCGACTATGAAGGCCTCAGTATGCTGTCGGGGCTTCCGGACGCCATGCGGGGCGGCCTGGTCCAGAACCTGTACAGCTTCGCTAAAGCGGACGCCGCGCCCGACCTGGTGCTCATGCTGGGCGCGCGCTTCGGCCTGAACACGGGCCACGGCAGCGGCCAGCTGATCCCGCACTCGGCCCAGGTCATCCAGGTCGACCCGGACGCGTGCGAGCTGGGCCGGCTGCAGGGCATCGCGCTTGGCATCGTCGCCGACGTGGGCGGCACCATCGAGGCCCTCGCGCAGGCCACCG CCCAGGACGCGGCGTGGCCCGACCGCGGCGACTGGTGCGCCAAGGTCACCGACCTCGCCCAGGAGCGCTACGCGTCGATCGCGGCCAAGTCCTCGTCCGAGCACGCCCTGCACCCCTTCCACGCCTCGCAGGTCATCGCCAAGCACGTCGACGCGGGCGTGACGGTCGTGGCCGACGGCGGGCTGACCTATCTGTGGCTGTCCGAGGTGATGAGCCGTGTGAAGCCGGGCGGCTTCCTCTGCCACGGCTACCTGAACTCCATGGGCGTCGGCTTCGGGACCGCGCTCGGCGCCCAGGTCGCCGACCTGGAGGCCGGCCGGCGCACGATCCTCGTGACCGGCGACGGCTCGGTGGGCTACTCCATCGGCGAGTTCGACACCCTGGTCCGGAAGCAGCTGCCGCTGATCGTGATCATCATGAACAACCAGTCGTGGGGCTGGACCCTCCACTTCCAGCAGCTCGCGGTCGGCCCGAACCGCGTGACCGGCACGCGCCTCGAGAACGGCTCGTACCACGGCGTCGCCGCGGCCTTCGGCGCCGACGGCTACCACGTCGACTCGGTCGAGTCGTTCAGCGCCGCCCTCGCCCAGGCCCTGGCCCACAACCGCCCGGCCTGCATCAACGTCGCGGTCGCGCTCGACCCCATCCCGCCGGAGGAGCTCATCCTCATCGGCATGGACCCGTTCGCCGGCAGCACCGAGAACCTCTACTTCCAGTCCGGCGCCTAA 12 Dak1 coding nucleic acid sequence ATGTCCGCTAAATCGTTTGAAGTCACAGATCCAGTCAATTCAAGTCTCAAAGGGTTTGCCCTTGCTAACCCCTCCATTACGCTGGTCCCTGAAGAAAAAATTCTCTTCAGAAAGACCGATTCCGACAAGATCGCATTAATTTCTGGTGGTGGTAGTGGACATGAACCTACACACGCCGGTTTCATTGGTAAGGGTATGTTGAGTGGCGCCGTGGTTGGCGAAATTTTTGCATCCCCTTCAACAAAACAGATTTTAAATGCAATCCGTTTAGTCAATGAAAATGCGTCTGGCGTTTTATTGATTGTGAAGAACTACACAGGTGATGTTTTGCATTTTGGTCTGTCCGCTGAGAGAGCAAGAGCCTTGGGTATTAACTGCCGCGTTGCTGTCATAGGTGATGATGTTGCAGTTGGCAGAGAAAAGGGTGGTATGGTTGGTAGAAGAGCATTGGCAGGTACCGTTTTGGTTCATAAGATTGTAGGTGCCTTCGCAGAAGAATATTCTAGTAAGTATGGCTTAGACGGTACAGCTAAAGTGGCTAAAATTATCAACGACAATTTGGTGACCATTGGATCTTCTTTAGACCATTGTAAAGTTCCTGGCAGGAAATTCGAAAGTGAATTAAACGAAAAACAAATGGAATTGGGTATGGGTATTCATAACGAACCTGGTGTGAAAGTTTTAGACCCTATTCCTTCTACCGAAGACTTGATCTCCAAGTATATGCTACCAAAACTATTGGATCCAAACGATAAGGATAGAGCTTTTGTAAAGTTTGATGAAGATGATGAAGTTGTCTTGTTAGTTAACAATCTCGGCGGTGTTTCTAATTTTGTTATTAGTTCTATCACTTCCAAAACTACGGATTTCTTAAAGGAAAATTACAACATAACCCCGGTTCAAACAATTGCTGGCACATTGATGACCTCCTTCAATGGTAATGGGTTCAGTATCACATTACTAAACGCCACTAAGGCTACAAAGGCTTTGCAATCTGATT TTGAGGAGATCAAATCAGTACTAGACTTGTTGAACGCATTTACGAACGCACCGGGCTGGCCAATTGCAGATTTTGAAAAGACTTCTGCCCCATCTGTTAACGATGACTTGTTACATAATGAAGTAACAGCAAAGGCCGTCGGTACCTATGACTTTGACAAGTTTGCTGAGTGGATGAAGAGTGGTGCTGAACAAGTTATCAAGAGCGAACCGCACATTACGGAACTAGACAATCAAGTTGGTGATGGTGATTGTGGTTACACTTTAGTGGCAGGAGTTAAAGGCATCACCGAAAACCTTGACAAGCTGTCGAAGGACTCATTATCTCAGGCGGTTGCCCAAATTTCAGATTTCATTGAAGGCTCAATGGGAGGTACTTCTGGTGGTTTATATTCTATTCTTTTGTCGGGTTTTTCACACGGATTAATTCAGGTTTGTAAATCAAAGGATGAACCCGTCACTAAGGAAATTGTGGCTAAGTCACTCGGAATTGCATTGGATACTTTATACAAATATACAAAGGCAAGGAAGGGATCATCCACCATGATTGATGCTTTAGAACCATTCGTTAAAGAATTTACTGCATCTAAGGATTTCAATAAGGCGGTAAAAGCTGCAGAGGAAGGTGCTAAATCCACTGCTACATTCGAGGCCAAATTTGGCAGAGCTTCGTATGTCGGCGATTCATCTCAAGTAGAAGATCCTGGTGCAGTAGGCCTATGTGAGTTTTTGAAGGGGGTTCAAAGCGCCTTGTAA

The strain was prepared by multi-insert cloning using an infusion enzyme, and Sac1 was used as a restriction enzyme. RBS was inserted in front of each gene using primers, and the primers used for cloning are shown in Table 4 below. The underlined sequence is RBS.

order
number designation order 13 Forward Primer 1 CCTGCAAGAGTGGGAATAAGAGCTC CTTAGACGAAAAAGGAGGTATTTTT ATGGCCATGATCACGGGCGGC 14 Reverse primer 1 GGACATTCCTTTAGGCGCCGGACTGGAAG 15 Forward primer 2 CGGCGCCTAA AGGA ATGTCCGCTAAATCGTTTGAAGT 16 Reverse primer 2 AGCAGCGGGGCTGGCTTAAGTTACAAGGCGCTTTGAACCC

1-3. Preparation of recombinant strains that have deleted the competitive pathway to MG

M. extorquens AM1 has a path from MG through lactoylglutathione to lactate. Therefore, the flow of MG produced in M. extorquens AM1 is divided into hydroxyacetone and lactate in the overexpressed 1,2-PDO production pathway. It deleted genes gloA working enzyme for converting lactose to galacto MG as one encoding a glutathione glutathione decomposing enzyme (Lactoylglutathione lyase) by that the flow losses occur in lactate M. extorquens AM1β gloA pMEV :: :: :: mgsA yqhD :: gldA strain (MPG101) and M. extorquens AM1β gloA :: pMEV :: mgsA :: yqhD :: gldA :: fls :: dak1 strain (MPG111) were developed. Since the two genes Mex_1p0359 and Mex_1p4683 of M. extorquens AM1 correspond to gloA, Mex_1p0359 and Mex_1p4683 were deleted using the pk19mobsacB vector.

The sequence of the pk19mobsacB vector is the same as the sequence of SEQ ID NO: 19, and the sequence of the related gene is shown in Table 5 below.

order
number designation order 17 Mex_1p0359
Coding nucleic acid sequence ATGGTTCGCGTGGCGGACCTCGACCGCGCGCTCGCCTTCTACGTCGATGCCTTCGGGCTCAAGGAGGTCCGGCGCGTCGAGAATGAGAAGGGCCGCTTCACCCTCGTCTTCCTCGCCGCTCCGGGGGATGTCGAGCGGGCCGAGGCGACGAAATCGCCGCTGATCGAGCTGACCTACAATTGGGATCCCGAGACCTATTCCGGCGGGCGCAACTTCGGGCACCTCGCCTATCAGGTCGATGACATCTACGCCTTCTGCCAGCGGCTGAAGATGCGGGCGTGACCATCAACCGGCCGCCGCGCGACGGGTACATGGCCTTCGTGCGCTCGCCCGACGGCATTTCCATCGAGATCCTGCAGAAGGGCGGGGCGAAGCCCCCGCAGGAGCCGTGGGCCTCCATGGAGAACACCGGAACCTGGTAG 18 Mex_1p4683
Coding nucleic acid sequence ATGGCCAAGCCCGTCCACACCATGATCCGCGTCCGCGACGAGGCGCGCTCGCGGGACTACTATGCCCGCGCCTTCGGCCTGGAGCCGGCCGACCGGTTCGACTTTCCGGACTTCACGCTGCTCTACCTGCGCGACCCATCCTCGCCGTTCGAACTCGAACTGACGGTCAACAAGGACCGCGCCGAGCCCTACAATCTCGGCGACGGCTACGGTCACCTCGCCTTCGTGGTGGAGGATGCCGAGGCCGAGCATGCCCGCTTCGAGCGCGAGGGGCTTCCGGTCACGCCGGTCAAAACCCTCAAGCACGGCGACACCGCGCTCGCGACCTTCTTTTTCGCCACCGACCCGGACGGCTACAAGATCGAGGTGATCCAGAAGGGCGGCCGTTTCGCCTGA

The strain was prepared by multi-insert cloning using Infusion enzyme. When Mex_1p0359 was deleted, Hind² and EcoRI were used as restriction enzymes, and upstream and downstream were designated as shown in Table 6 below.

order
number designation order 20 Mex_1p0359_upstream ATGAACACGGACGATTTCGCCTACGATTTCGACGCGCCCGATTCGGACTGGAACTACGAGCCGTCGCGGGCCGATCTGTTCCGGCAGATCGACGCGCCACTCTATACCACTGATTCCAACGGCTGGCTGACCTACTACAACGAAGCCGCCGCACAGCTTTGGGGATTCCGCCCCGTGATCGGCAAGGCGCGCTGGTGCGGCGCTTGGCGGCTGTTCGAGGCGGACGGCGCGCCGCTGCCGCACGACCTGTCGCCGATGGCGCTCACCCTCAAAGGCGCGCGCCCAGTGCGCGGCGTCCAGATCGGCCTCGAACGGCCGGACGGAAGCCGGATGGCGTTCCTGCCTTACCCGACGGCGTTGCGCGACGGGACGGGCGCCGTGGTGGGGGGATGCAACATCCTCCTCGCCGTCGAGCGCTCAAGCCTGCGCGTTCCGCTCCGCGGGGCGCTCCAGGGCCGGCCGACGCTTCGGGCCGCCCAACTTGCGAGCATGCACAGATGTTCGGCGTGAGCTTGCCGCGTCTGCCTCGGCGATCCACCTCGTTCGAGCGAAGCGATCCGCGGGATGTCTCCCCGGATCGCCTTGCCGCCTGAGGAACGTGCCGCCTGAAAAACAGGAGGAAGCCATGTCGGTCGATACCAACTGGAGCCTCGACGCCGTCCAGAGCCTGCGCAGCATGGCCCGCGAGGGCATACCCCTCTCCGTCATCAGCCTGCGGCTCAAGCGGCCGGTCGATGCGGTCTGCGCCAAGCTCGCCGAACTCGGCATCACGCCCAAGCTCGAGCTTTGACCGGACGGGCGCCGGGGCATCGCGCCCGGGCGCCCACCCGCCGGAC 21 Mex_1p0359 _downstream CGTGTGCAAGTATTCCATGGCCACACCGGATGATGACAGGGATGTCCGGCTATCTGGCCCGGCACGCAAACCCGTCAACGGGGCGACCGATCAGGCTCCCGCGCGGCCAGTCAGCCAGCGGCGGCCAAACGGCCGTTCGACCGGCGGCGCCCGCCGGATTCGACCGGACTGCCGGTGAATAACATCCATTTGAAAAAAGCTTCTCTTGTGGCAATTGCTTGATCCAATTACGATCACTATCGCGTCTCGCAAGCTTTCCGTTAAATTATAGTGACATGGCATTGAAAAATAGAAGGCATTCCTTCGCAACTGACTTGGCGTCGATGCATATTATAGCATTTAAGAGCTTTTGTTTCCCGAAATGGCAACAGCTAAGATTCGCCGCGTCCCGGCCTGCGATGATCGTCGAATCGGGCATGCCGCGACCGGGAGACGGCGC

Primers used to insert upstream and downstream into pK19mobsacB (SEQ ID NOs: 22 to 25), primers used to select colonies in which a single recombinant has occurred in the upstream and downstream, respectively (SEQ ID NOs: 26 to 29 ) And primers (SEQ ID NOs: 30 and 31) used in PCR performed to distinguish between mutant and wild type are shown in Table 7 below.

order
number designation order 22 Forward Primer 1 GACCATGATTACGCCAAGCTTATGAACACGGACGATTTCGCC 23 Reverse primer 1 CTTGCACACGGTCCGGCGGGTGGGCGCC 24 Forward primer 2 CCCGCCGGACCGTGTGCAAGTATTCCATGGCC 25 Reverse primer 2 AAAACGACGGCCAGTGAATTCGCGCCGTCTCCCGGTCGC 26 Forward Primer 3 AACCCGCAGAAACACGCTTT 27 Reverse primer 3 GAAGCTAGCTTATCGCGCCA 28 Forward Primer 4 TTTCTGCGCGTAATCTGCTG 29 Reverse primer 4 ACGATGATGGCGAGGGTG 30 Forward Primer 5 GGAGCCTGATGAACACGGAC 31 Reverse primer 5 GCAATCATGCGCCGTCTC

For a single recombinant colony, the desired mutation was finally selected through PCR through sucrose selection and kanamycin selection. When Mex_1p4683 was deleted, Hind²EcoRI was used as a restriction enzyme, and the upstream and downstream were as follows: It was designated as in Table 8.

order
number designation order 32 Mex_1p0359_upstream GCTGGTATCTGCCGCCGCGCTGCCGGCAGAACCTCCGGAAAGACCCATTGTCGCCCCCTGTCGTTTCCGGCTTGACCGTGGACGAGATTCGCTCCCGAGAGCGTGAGGGAACGCACGCGACCTGTCGCGTCCGGCGCGATCGCCGGATCGGGCGGGATGCGGTGCCGTTCTGAAGTGCTGTGAGGGAGAACATTCGATGCGCTCAGGGGCCGCTTCCGTCTTCTTCCCCCTGTTCGGGCTCGTGCTCACGGTCGGCGCGGCCGATGCGCAGACGCCGCCCGAGCCAGGCGCTGCCCGGCCGCTGGTGATCCCCCAGGTCGAGGGCGAGCGCCAGGGCAAGGTGCTGGGCCGGGCGCTCGCCTGCGGCGCGGAGCGCGAGCGGGTCGATCGGGTGCTGAGAGCCGGCCGCGAGCGCATGATGGCGGCGGTCGGCCGGGCGCTCACGGAGGAGCGCTACGCCCTGGCGCTCGACGACGCGATGCGTCTGGAGACGAGCCTGCCTGCGCCCTCTTCGATCGCGTGCGAGAAGGCGCTGGCCGGGCTCGAACGCCTGGAGAAGGCGCCGTAGACACCCAAACGA 33 Mex_1p0359_downstream CGGCCCTCTCTCCCCGGTACGGATGTTGTGTGGATGAGCGCCCATCTGGCGCAGTCCGGCCGGGCGGAAAGGGTTTTTGCCGTTTACCCTTCGTCAACCTTACCGGGTGCTTGCTGCCGGCAATGATGCGCACCGCGTCCCTCACCGCCGTCGATCCCGAAGCCCTGCTGCCGCCGGGCGACGCCGCCCTGCGCGCGGCGATGCGGGGCTGGCGCGAGGCGCTGGCGCGGGAGCGGCGCATGGCCGCCAACACGGTCGAGGCCTACGAGCGTGACCTGCGCCAGTTTCTGATCCATCGCGCCGCCCGCTCCGGCACGCCCACCATCGCCGGGCTGATCGCCCTGAAGCCCCGCGACCTGCGCGCCTTCATGGCCGCGCGCCGGGCCGAGGGGATCGGTGGGCGCTCCCTGATGCGGATGCTGGCGGGTCTGCGCTCCTTCGCCCGCTTCCTCGAACGGGAGGGACACGGCAGCGTCGCGGCGCTCGGCGCGGTGCGCTCCCCCAAGGTCGAGCGCCGCCTGCCGCGCCCGCTTCCCATTTCCGCCGCGCTGGCGATGACCGCGCCCGAGACCCGCCCCGACGACGACCGCGCCCCGTGGGTGCTCGCCCGCGACGCGGCGGTGATCGCCCTCCTCTACGGCTCGGGCCTGCGCATCTCGGAAGCCTTAGGGCTCACCGCCCGCGACGCGCCGATGCCGGGCATCGACGAGGTGCGGGTGACGGGCAAGGGTGGGAAGGTCCGCGCCGTGCCGGTGCTGCCGGCGGTGGCCGAGGCGGTGGCCGCCTACCTGTCGCTGTGCCCGCACCCCCTCGATCCGGAGGGGCCGCTCTTCGTCGGCGTGAAGGGCGGGCCGCTCTCGCCGCGAGTGGTCCAGTACGCGGTTTCCGCGCTCCGCGGCGCGCTCGGCCTGCCCGAGAGCGCGACCCCGCACGCCCTGCGACACTCCTTCGCAACCCATCTGCTCGCCCGCCGGGGCGAGCTGCGGGCGATCCAGGAATT GCTCGGCCACGCCTCGCTCTCGACCACGCAAATCTACACCAAGGTCGATGCCGCCCGCCTGATGAGCGCCTTCGAGGACGCCCATCCCCGCGCCCGGCGGCTGCCGCCCTCGGAACCGCCGTCACCGCGTTCCGAAACCGTTGATGCAGAGAGCAAGGCACGAGCGCCCCCGATCCGGCTAGGCGCGCGAGGCCGGCTAGGCAGGT

The primers used to insert upstream and downstream into pK19mobsacB, the primers used to select colonies in which single recombination has occurred in each of the upstream and downstream, and the primers used in PCR to distinguish between mutant and wild type are shown in Table 9 below.

order
number designation order 34 Forward Primer 1 GACCATGATTACGCCAAGCTTGCTGGTATCTGCCGCCGC 35 Reverse primer 1 GAGAGGGCCGTCGTTTGGGTGTCTACGGCG 36 Forward primer 2 ACCCAAACGACGGCCCTCTCTCCCCGGTAC 37 Reverse primer 2 AAAACGACGGCCAGTGAATTCTCACGCATTTCTACGCTCCACC 38 Forward Primer 3 AGATCGAGACGAGAAGCG 39 Reverse primer 3 CCACACAACATCCGTACC 40 Forward Primer 4 GACCATGATTACGCCAAGC 41 Reverse primer 4 CCACACAACATCCGTACC 42 Forward Primer 5 CAGAACCTCCGGAAAGACCC 43 Reverse primer 5 TCCACACAACATCCGTACCG

Example 2: Confirmation of production amount from recombinant strain

The strains prepared in Example 1 (MPG001, MPG011, MPG101 and MPG111) contained 100 ml of NMS medium in a 500 ml baffle flask and were cultured at 30° C. rpm 200. The composition of the medium is shown in Table 10 below.

NMS stock solution recipe Adding amount for 1L MeOH 125mM MgSO ₄ 7H ₂ O Stock 1g KNO ₃ Stock 1g CaCl ₂ 2H ₂ O Stock 0.228g 3.8% (w/v) solution Fe-EDTA 0.0038g 0.1% (w/v) NaMo H ₂ O 0.0006g FeSO4·H2O 0.0005g ZnSO4·H2O 0.0004g MnCl2·H2O 0.00002g CoCl2·H2O 0.00005g NiCl2·H2O 0.00001g H3BO3 (boric acid) 0.000015g EDTA 0.00025g KH2PO4 0.26g Na2HPO4·(H2O) 0.62g Biotin 0.02mg Folic acid 0.02mg Thiamine HCl 0.05mg Ca pantothenate 0.05mg Vitamin B12 0.001mg Riboflavin 0.05mg Nicotiamide 0.05mg CuSO4 5H2O 2.5mg

Methylobacterium extorkens AM1 grows and forms a clump, making it difficult to accurately measure OD above a certain OD. Therefore, only some significant data were indicated for OD data. The 1,2-PDO produced from the flask fermentation result was confirmed and shown in FIGS. 3 to 6 and Table 11.

MPG001 MPG011 MPG101 MPG111 Average(mg/L) 8.6 9.8 41.1 58.8 Max(mg/L) 10.1 14.2 52.9 61.6

As can be seen in FIGS. 3 to 6 and Table 12, strain MPG111, which improved DHAP flow for 1,2-PDO-producing strain MPG001 and prevented the flow from MG to lactate, a competitive pathway, improved about 6.8 times. -PDO production was shown. Compared with the increase in the production of 1,2-PDO due to overexpression of formolease and dihydroxyacetone kinase, the production of 1,2-PDO due to deletion of the gloA coding gene It could be determined that the degree of increase was remarkably high.

<110> Industry-University Cooperation Foundation Sogang University <120> Methylotroph with enhanced 1,2-propylene glycol productivity and method for producing 1,2-propylene glycol using the same <130> PN190334 <160> 43 <170> KoPatentIn 3.0 <210> 1 <211> 5730 <212> DNA <213> Artificial Sequence <220> <223> pMEV vector <400> 1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg 60 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc 120 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca 180 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc 240 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt 300 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg 360 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca 420 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt 480 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac 540 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc 600 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc 660 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg 720 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg 780 tgagcgtggg tctcgcggta tcattgcagc accagtactg accccgtaga aaagatcaaa 840 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 900 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 960 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 1020 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 1080 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 1140 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 1200 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 1260 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 1320 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 1380 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 1440 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 1500 tttcctgcgt tatcccctga ttctgtggat aaccgtagca tgcgttgacg acaacggtgc 1560 gatgggtccc ggccccggtc aagacgatgc caatacgttg cgacactacg ccttggcact 1620 tttagaattg ccttatcgtc ctgataagaa atgtccgacc agctaaagac atcgcgtcca 1680 atcaaagcct agaaaatata ggcgaaggga cgctaatggg cccttcacac agaggagaga 1740 cagcgaattc tggtaccttc tagactatcg actagttggt acctctgcag aagcttccgg 1800 agctcttaag ccagccccgc tgctcccggc atccgcttac agataaaacg aaaggctcag 1860 tcgaaagact gggcctttcg ttttatacaa gccctccagg ggagatgcgt ggatccttag 1920 atctaggcct gaatcgcccc atcatccagc cagaaagtga gggagccacg gttgatgaga 1980 gctttgttgt aggtggacca gttggtgatt ttgaactttt gctttgccac ggaacggtct 2040 gcgttgtcgg gaagatgcgt gatctgatcc ttcaactcag caaaagttcg atttattcaa 2100 caaagccgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac caattaacca 2160 attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat 2220 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc 2280 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa 2340 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag 2400 tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa 2460 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc 2520 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag 2580 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat 2640 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc 2700 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca 2760 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt 2820 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt 2880 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta 2940 atcgcggcct cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac 3000 tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt 3060 aacatcagag attttgagac acaacgtggc tttccccccc ccccctgcag gtccgacacg 3120 gggatggatg gcgttcccga tcatggtcct gcttgcttcg ggtggcatcg gaatgccggc 3180 gctgcaagca atgttgtcca ggcaggtgga tgaggaacgt caggggcagc tgcaaggctc 3240 actggcggcg ctcaccagcc tgacctcgat cgtcggaccc ctcctcttca cggcgatcta 3300 tgcggcttct ataacaacgt ggaacgggtg ggcatggatt gcaggcgctg ccctctactt 3360 gctctgcctg ccggcgctgc gtcgcgggct ttggagcggc gcagggcaac gagccgatcg 3420 ctgatcgtgg aaacgatagg cctatgccat gcgggtcaag gcgacttccg gcaagctata 3480 cgcgccctag aattgtcaat tttaatcctc tgtttatcgg cagttcgtag agcgcgccgt 3540 gcgtcccgag cgatactgag cgaagcaagt gcgtcgagca gtgcccgctt gttcctgaaa 3600 tgccagtaaa gcgctggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg 3660 tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa 3720 ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc 3780 gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata 3840 gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt 3900 gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga 3960 cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg 4020 cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc 4080 ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa 4140 cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca 4200 caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt 4260 gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt 4320 ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg 4380 cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa 4440 cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt 4500 cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc 4560 ctgttcgaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc 4620 cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg agccgacgga 4680 ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc 4740 ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt 4800 cattcaccct ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc cttattcctg 4860 atttgacccg cctggtgcct tggtgtccag ataatccacc ttatcggcaa tgaagtcggt 4920 cccgtagacc gtctggccgt ccttctcgta cttggtattc cgaatcttgc cctgcacgaa 4980 taccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac 5040 gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca 5100 cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga 5160 ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca 5220 ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt 5280 ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc tacaggctca 5340 tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct 5400 tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat 5460 ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca 5520 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta 5580 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa 5640 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga 5700 agcagggtta tgcagcggaa aagatccgtc 5730 <210> 2 <211> 414 <212> DNA <213> Bacillus subtilis <400> 2 atgaaaattg ctttgatcgc gcatgacaag aaaaaacagg atatggttca atttacgact 60 gcctatcggg atattttaaa gaatcatgat ctatacgcaa ccggaaccac agggttgaaa 120 attcatgagg cgacaggtct tcaaattgaa cgttttcaat ccggcccttt agggggagac 180 cagcaaatcg gtgcactgat cgctgccaat gcactcgatc ttgtcatttt tttgcgcgac 240 ccgctgaccg cgcagccgca tgaaccggat gtctcggcat taatccgttt atgtgatgtg 300 tattccattc cgctcgccac aaatatgggt actgcggaaa ttcttgtgcg cacacttgat 360 gaaggtgttt tcgaattccg tgaccttctt cggggagaag agccgaatgt ataa 414 <210> 3 <211> 1164 <212> DNA <213> Escherichia coli <400> 3 atgaacaact ttaatctgca caccccaacc cgcattctgt ttggtaaagg cgcaatcgct 60 ggtttacgcg aacaaattcc tcacgatgct cgcgtattga ttacctacgg cggcggcagc 120 gtgaaaaaaa ccggcgttct cgatcaagtt ctggatgccc tgaaaggcat ggacgtgctg 180 gaatttggcg gtattgagcc aaacccggct tatgaaacgc tgatgaacgc cgtgaaactg 240 gttcgcgaac agaaagtgac tttcctgctg gcggttggcg gcggttctgt actggacggc 300 accaaattta tcgccgcagc ggctaactat ccggaaaata tcgatccgtg gcacattctg 360 caaacgggcg gtaaagagat taaaagcgcc atcccgatgg gctgtgtgct gacgctgcca 420 gcaaccggtt cagaatccaa cgcaggcgcg gtgatctccc gtaaaaccac aggcgacaag 480 caggcgttcc attctgccca tgttcagccg gtatttgccg tgctcgatcc ggtttatacc 540 tacaccctgc cgccgcgtca ggtggctaac ggcgtagtgg acgcctttgt acacaccgtg 600 gaacagtatg ttaccaaacc ggttgatgcc aaaattcagg accgtttcgc agaaggcatt 660 ttgctgacgc taatcgaaga tggtccgaaa gccctgaaag agccagaaaa ctacgatgtg 720 cgcgccaacg tcatgtgggc ggcgactcag gcgctgaacg gtttgattgg cgctggcgta 780 ccgcaggact gggcaacgca tatgctgggc cacgaactga ctgcgatgca cggtctggat 840 cacgcgcaaa cactggctat cgtcctgcct gcactgtgga atgaaaaacg cgataccaag 900 cgcgctaagc tgctgcaata tgctgaacgc gtctggaaca tcactgaagg ttccgatgat 960 gagcgtattg acgccgcgat tgccgcaacc cgcaatttct ttgagcaatt aggcgtgccg 1020 acccacctct ccgactacgg tctggacggc agctccatcc cggctttgct gaaaaaactg 1080 gaagagcacg gcatgaccca actgggcgaa aatcatgaca ttacgttgga tgtcagccgc 1140 cgtatatacg aagccgcccg ctaa 1164 <210> 4 <211> 1104 <212> DNA <213> Escherichia coli <400> 4 atggaccgca ttattcaatc accgggtaaa tacatccagg gcgctgatgt gattaatcgt 60 ctgggcgaat acctgaagcc gctggcagaa cgctggttag tggtgggtga caaatttgtt 120 ttaggttttg ctcaatccac tgtcgagaaa agctttaaag atgctggact ggtagtagaa 180 attgcgccgt ttggcggtga atgttcgcaa aatgagatcg accgtctgcg tggcatcgcg 240 gagactgcgc agtgtggcgc aattctcggt atcggtggcg gaaaaaccct cgatactgcc 300 aaagcactgg cacatttcat gggtgttccg gtagcgatcg caccgactat cgcctctacc 360 gatgcaccgt gcagcgcatt gtctgttatc tacaccgatg agggtgagtt tgaccgctat 420 ctgctgttgc caaataaccc gaatatggtc attgtcgaca ccaaaatcgt cgctggcgca 480 cctgcacgtc tgttagcggc gggtatcggc gatgcgctgg caacctggtt tgaagcgcgt 540 gcctgctctc gtagcggcgc gaccaccatg gcgggcggca agtgcaccca ggctgcgctg 600 gcactggctg aactgtgcta caacaccctg ctggaagaag gcgaaaaagc gatgcttgct 660 gccgaacagc atgtagtgac tccggcgctg gagcgcgtga ttgaagcgaa cacctatttg 720 agcggtgttg gttttgaaag tggtggtctg gctgcggcgc acgcagtgca taacggcctg 780 accgctatcc cggacgcgca tcactattat cacggtgaaa aagtggcatt cggtacgctg 840 acgcagctgg ttctggaaaa tgcgccggtg gaggaaatcg aaaccgtagc tgcccttagc 900 catgcggtag gtttgccaat aactctcgct caactggata ttaaagaaga tgtcccggcg 960 aaaatgcgaa ttgtggcaga agcggcatgt gcagaaggtg aaaccattca caacatgcct 1020 ggcggcgcga cgccagatca ggtttacgcc gctctgctgg tagccgacca gtacggtcag 1080 cgtttcctgc aagagtggga ataa 1104 <210> 5 <211> 68 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 5 gagagacagc gaattcccgc gagggaattc ccaaaggcca ggtattttat gaaaattgct 60 ttgatcgc 68 <210> 6 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 6 tgggtctatt gggagttata cattcggctc ttctccc 37 <210> 7 <211> 54 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 7 ctcccaatag acccaaaggc ggtagagaaa tgaacaactt taatctgcac accc 54 <210> 8 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 8 ttgcgcttag ttgccttagc gggcggcttc gtatata 37 <210> 9 <211> 51 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 9 ggcaactaag cgcaaagatc gaggtaataa atggaccgca ttattcaatc a 51 <210> 10 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 10 cggggctggc ttaagagctc ttattcccac tcttgcagga a 41 <210> 11 <211> 1728 <212> DNA <213> Artificial Sequence <220> <223> fls coding sequences <400> 11 atggccatga tcacgggcgg cgagctggtc gtgcgcacgc tgatcaaggc gggtgtcgag 60 cacctctttg gtctccacgg tatccacatc gacaccatct tccaggcctg cctggaccat 120 gatgtcccga tcatcgacac ccgccacgag gcggcagccg gacatgcggc ggaaggctac 180 gcccgcgccg gcgccaagct cggcgtcgcg cttgtgacgg cgggcggcgg cttcaccaac 240 gcggtgaccc cgatcgccaa tgcccgcacc gaccgcacgc cggtcctctt ccttaccggc 300 tcgggcgccc tcagggatga cgagacgaac accctccagg cggggatcga ccaggtcgcg 360 atggccgccc ctatcaccaa gtgggcccat cgggtaatgg ccaccgaaca catccctcgg 420 ctggtcatgc aggcgattcg cgccgcgctg agcgcccccc gcggcccagt tctgctcgac 480 ctcccgtggg acatcctcat gaaccagatt gacgaagata gcgtcatcat cccggacctc 540 gtgctgtctg cccacggcgc gcaccccgat cccgcggacc tcgatcaggc cctggcgctg 600 ctccgcaagg ccgagcgtcc ggttatcgtt ctgggcagtg aggccagccg cacggcccga 660 aagacggccc tctcggcctt cgtcgcggcc acgggcgtgc cggtcttcgc cgactatgaa 720 ggcctcagta tgctgtcggg gcttccggac gccatgcggg gcggcctggt ccagaacctg 780 tacagcttcg ctaaagcgga cgccgcgccc gacctggtgc tcatgctggg cgcgcgcttc 840 ggcctgaaca cgggccacgg cagcggccag ctgatcccgc actcggccca ggtcatccag 900 gtcgacccgg acgcgtgcga gctgggccgg ctgcagggca tcgcgcttgg catcgtcgcc 960 gacgtgggcg gcaccatcga ggccctcgcg caggccaccg cccaggacgc ggcgtggccc 1020 gaccgcggcg actggtgcgc caaggtcacc gacctcgccc aggagcgcta cgcgtcgatc 1080 gcggccaagt cctcgtccga gcacgccctg caccccttcc acgcctcgca ggtcatcgcc 1140 aagcacgtcg acgcgggcgt gacggtcgtg gccgacggcg ggctgaccta tctgtggctg 1200 tccgaggtga tgagccgtgt gaagccgggc ggcttcctct gccacggcta cctgaactcc 1260 atgggcgtcg gcttcgggac cgcgctcggc gcccaggtcg ccgacctgga ggccggccgg 1320 cgcacgatcc tcgtgaccgg cgacggctcg gtgggctact ccatcggcga gttcgacacc 1380 ctggtccgga agcagctgcc gctgatcgtg atcatcatga acaaccagtc gtggggctgg 1440 accctccact tccagcagct cgcggtcggc ccgaaccgcg tgaccggcac gcgcctcgag 1500 aacggctcgt accacggcgt cgccgcggcc ttcggcgccg acggctacca cgtcgactcg 1560 gtcgagtcgt tcagcgccgc cctcgcccag gccctggccc acaaccgccc ggcctgcatc 1620 aacgtcgcgg tcgcgctcga ccccatcccg ccggaggagc tcatcctcat cggcatggac 1680 ccgttcgccg gcagcaccga gaacctctac ttccagtccg gcgcctaa 1728 <210> 12 <211> 1755 <212> DNA <213> Artificial Sequence <220> <223> dak1 coding sequences <400> 12 atgtccgcta aatcgtttga agtcacagat ccagtcaatt caagtctcaa agggtttgcc 60 cttgctaacc cctccattac gctggtccct gaagaaaaaa ttctcttcag aaagaccgat 120 tccgacaaga tcgcattaat ttctggtggt ggtagtggac atgaacctac acacgccggt 180 ttcattggta agggtatgtt gagtggcgcc gtggttggcg aaatttttgc atccccttca 240 acaaaacaga ttttaaatgc aatccgttta gtcaatgaaa atgcgtctgg cgttttattg 300 attgtgaaga actacacagg tgatgttttg cattttggtc tgtccgctga gagagcaaga 360 gccttgggta ttaactgccg cgttgctgtc ataggtgatg atgttgcagt tggcagagaa 420 aagggtggta tggttggtag aagagcattg gcaggtaccg ttttggttca taagattgta 480 ggtgccttcg cagaagaata ttctagtaag tatggcttag acggtacagc taaagtggct 540 aaaattatca acgacaattt ggtgaccatt ggatcttctt tagaccattg taaagttcct 600 ggcaggaaat tcgaaagtga attaaacgaa aaacaaatgg aattgggtat gggtattcat 660 aacgaacctg gtgtgaaagt tttagaccct attccttcta ccgaagactt gatctccaag 720 tatatgctac caaaactatt ggatccaaac gataaggata gagcttttgt aaagtttgat 780 gaagatgatg aagttgtctt gttagttaac aatctcggcg gtgtttctaa ttttgttatt 840 agttctatca cttccaaaac tacggatttc ttaaaggaaa attacaacat aaccccggtt 900 caaacaattg ctggcacatt gatgacctcc ttcaatggta atgggttcag tatcacatta 960 ctaaacgcca ctaaggctac aaaggctttg caatctgatt ttgaggagat caaatcagta 1020 ctagacttgt tgaacgcatt tacgaacgca ccgggctggc caattgcaga ttttgaaaag 1080 acttctgccc catctgttaa cgatgacttg ttacataatg aagtaacagc aaaggccgtc 1140 ggtacctatg actttgacaa gtttgctgag tggatgaaga gtggtgctga acaagttatc 1200 aagagcgaac cgcacattac ggaactagac aatcaagttg gtgatggtga ttgtggttac 1260 actttagtgg caggagttaa aggcatcacc gaaaaccttg acaagctgtc gaaggactca 1320 ttatctcagg cggttgccca aatttcagat ttcattgaag gctcaatggg aggtacttct 1380 ggtggtttat attctattct tttgtcgggt ttttcacacg gattaattca ggtttgtaaa 1440 tcaaaggatg aacccgtcac taaggaaatt gtggctaagt cactcggaat tgcattggat 1500 actttataca aatatacaaa ggcaaggaag ggatcatcca ccatgattga tgctttagaa 1560 ccattcgtta aagaatttac tgcatctaag gatttcaata aggcggtaaa agctgcagag 1620 gaaggtgcta aatccactgc tacattcgag gccaaatttg gcagagcttc gtatgtcggc 1680 gattcatctc aagtagaaga tcctggtgca gtaggcctat gtgagttttt gaagggggtt 1740 caaagcgcct tgtaa 1755 <210> 13 <211> 71 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 13 cctgcaagag tgggaataag agctccttag acgaaaaagg aggtattttt atggccatga 60 tcacgggcgg c 71 <210> 14 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 14 ggacattcct ttaggcgccg gactggaag 29 <210> 15 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 15 cggcgcctaa aggaatgtcc gctaaatcgt ttgaagt 37 <210> 16 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 16 agcagcgggg ctggcttaag ttacaaggcg ctttgaaccc 40 <210> 17 <211> 422 <212> DNA <213> Methylobacterium extorquens <400> 17 atggttcgcg tggcggacct cgaccgcgcg ctcgccttct acgtcgatgc cttcgggctc 60 aaggaggtcc ggcgcgtcga gaatgagaag ggccgcttca ccctcgtctt cctcgccgct 120 ccgggggatg tcgagcgggc cgaggcgacg aaatcgccgc tgatcgagct gacctacaat 180 tgggatcccg agacctattc cggcgggcgc aacttcgggc acctcgccta tcaggtcgat 240 gacatctacg ccttctgcca gcggctgaag atgcgggcgt gaccatcaac cggccgccgc 300 gcgacgggta catggccttc gtgcgctcgc ccgacggcat ttccatcgag atcctgcaga 360 agggcggggc gaagcccccg caggagccgt gggcctccat ggagaacacc ggaacctggt 420 ag 422 <210> 18 <211> 396 <212> DNA <213> Methylobacterium extorquens <400> 18 atggccaagc ccgtccacac catgatccgc gtccgcgacg aggcgcgctc gcgggactac 60 tatgcccgcg ccttcggcct ggagccggcc gaccggttcg actttccgga cttcacgctg 120 ctctacctgc gcgacccatc ctcgccgttc gaactcgaac tgacggtcaa caaggaccgc 180 gccgagccct acaatctcgg cgacggctac ggtcacctcg ccttcgtggt ggaggatgcc 240 gaggccgagc atgcccgctt cgagcgcgag gggcttccgg tcacgccggt caaaaccctc 300 aagcacggcg acaccgcgct cgcgaccttc tttttcgcca ccgacccgga cggctacaag 360 atcgaggtga tccagaaggg cggccgtttc gcctga 396 <210> 19 <211> 5722 <212> DNA <213> Artificial Sequence <220> <223> pk19mobsacB vector <400> 19 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 60 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 120 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca 180 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag 240 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgcatgcct 300 gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcactggcc gtcgttttac 360 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc 420 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc 480 gcagcctgaa tggcgaatgg cgcgataagc tagcttcacg ctgccgcaag cactcagggc 540 gcaagggctg ctaaaggaag cggaacacgt agaaagccag tccgcagaaa cggtgctgac 600 cccggatgaa tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa 660 agcaggtagc ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag 720 caagcgaacc ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag 780 taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca agatctgatc 840 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 900 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 960 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 1020 acctgtccgg tgccctgaat gaactccaag acgaggcagc gcggctatcg tggctggcca 1080 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 1140 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 1200 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 1260 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 1320 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 1380 ccaggctcaa ggcgcggatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 1440 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1500 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1560 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1620 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc 1680 tagaggatcg atccttttta acccatcaca tatacctgcc gttcactatt atttagtgaa 1740 atgagatatt atgatatttt ctgaattgtg attaaaaagg caactttatg cccatgcaac 1800 agaaactata aaaaatacag agaatgaaaa gaaacagata gattttttag ttctttaggc 1860 ccgtagtctg caaatccttt tatgattttc tatcaaacaa aagaggaaaa tagaccagtt 1920 gcaatccaaa cgagagtcta atagaatgag gtcgaaaagt aaatcgcgcg ggtttgttac 1980 tgataaagca ggcaagacct aaaatgtgta aagggcaaag tgtatacttt ggcgtcaccc 2040 cttacatatt ttaggtcttt ttttattgtg cgtaactaac ttgccatctt caaacaggag 2100 ggctggaaga agcagaccgc taacacagta cataaaaaag gagacatgaa cgatgaacat 2160 caaaaagttt gcaaaacaag caacagtatt aacctttact accgcactgc tggcaggagg 2220 cgcaactcaa gcgtttgcga aagaaacgaa ccaaaagcca tataaggaaa catacggcat 2280 ttcccatatt acacgccatg atatgctgca aatccctgaa cagcaaaaaa atgaaaaata 2340 tcaagtttct gaatttgatt cgtccacaat taaaaatatc tcttctgcaa aaggcctgga 2400 cgtttgggac agctggccat tacaaaacgc tgacggcact gtcgcaaact atcacggcta 2460 ccacatcgtc tttgcattag ccggagatcc taaaaatgcg gatgacacat cgatttacat 2520 gttctatcaa aaagtcggcg aaacttctat tgacagctgg aaaaacgctg gccgcgtctt 2580 taaagacagc gacaaattcg atgcaaatga ttctatccta aaagaccaaa cacaagaatg 2640 gtcaggttca gccacattta catctgacgg aaaaatccgt ttattctaca ctgatttctc 2700 cggtaaacat tacggcaaac aaacactgac aactgcacaa gttaacgtat cagcatcaga 2760 cagctctttg aacatcaacg gtgtagagga ttataaatca atctttgacg gtgacggaaa 2820 aacgtatcaa aatgtacagc agttcatcga tgaaggcaac tacagctcag gcgacaacca 2880 tacgctgaga gatcctcact acgtagaaga taaaggccac aaatacttag tatttgaagc 2940 aaacactgga actgaagatg gctaccaagg cgaagaatct ttatttaaca aagcatacta 3000 tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa 3060 acgcacggct gagttagcaa acggcgctct cggtatgatt gagctaaacg atgattacac 3120 actgaaaaaa gtgatgaaac cgctgattgc atctaacaca gtaacagatg aaattgaacg 3180 cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc actgactccc gcggatcaaa 3240 aatgacgatt gacggcatta cgtctaacga tatttacatg cttggttatg tttctaattc 3300 tttaactggc ccatacaagc cgctgaacaa aactggcctt gtgttaaaaa tggatcttga 3360 tcctaacgat gtaaccttta cttactcaca cttcgctgta cctcaagcga aaggaaacaa 3420 tgtcgtgatt acaagctata tgacaaacag aggattctac gcagacaaac aatcaacgtt 3480 tgcgccgagc ttcctgctga acatcaaagg caagaaaaca tctgttgtca aagacagcat 3540 ccttgaacaa ggacaattaa cagttaacaa ataaaaacgc aaaagaaaat gccgatgggt 3600 accgagcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac 3660 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccc tcgcggacgt 3720 gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc aggtaggccg 3780 acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg tctggaaggc 3840 agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca gccatccgct 3900 tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga ttcccgttga 3960 gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg ggtgggccta 4020 cttcacctat cctgccccgc tgacgccgtt ggatacacca aggaaagtct acacgaaccc 4080 tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa atcgctataa 4140 tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt ttgtggaata 4200 tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac aggcgagaga 4260 cgatgccaaa gagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt 4320 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttttcgccaa 4380 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag 4440 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta 4500 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctc 4560 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa 4620 agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct 4680 tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag 4740 gtatttattc ggcgcaaagt gcgtcgggtg atgctgccaa cttactgatt tagtgtatga 4800 tggtgttttt gaggtgctcc agtggcttct gtttctatca gggctggatg atcctccagc 4860 gcggggatct catgctggag ttcttcgccc accccaaaag gatctaggtg aagatccttt 4920 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 4980 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 5040 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 5100 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 5160 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 5220 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 5280 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 5340 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt 5400 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 5460 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 5520 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 5580 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 5640 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 5700 cctttgagtg agctgatacc gc 5722 <210> 20 <211> 836 <212> DNA <213> Methylobacterium extorquens <400> 20 atgaacacgg acgatttcgc ctacgatttc gacgcgcccg attcggactg gaactacgag 60 ccgtcgcggg ccgatctgtt ccggcagatc gacgcgccac tctataccac tgattccaac 120 ggctggctga cctactacaa cgaagccgcc gcacagcttt ggggattccg ccccgtgatc 180 ggcaaggcgc gctggtgcgg cgcttggcgg ctgttcgagg cggacggcgc gccgctgccg 240 cacgacctgt cgccgatggc gctcaccctc aaaggcgcgc gcccagtgcg cggcgtccag 300 atcggcctcg aacggccgga cggaagccgg atggcgttcc tgccttaccc gacggcgttg 360 cgcgacggga cgggcgccgt ggtgggggga tgcaacatcc tcctcgccgt cgagcgctca 420 agcctgcgcg ttccgctccg cggggcgctc cagggccggc cgacgcttcg ggccgcccaa 480 cttgcgagca tgcacagatg ttcggcgtga gcttgccgcg tctgcctcgg cgatccacct 540 cgttcgagcg aagcgatccg cgggatgtct ccccggatcg ccttgccgcc tgaggaacgt 600 gccgcctgaa aaacaggagg aagccatgtc ggtcgatacc aactggagcc tcgacgccgt 660 ccagagcctg cgcagcatgg cccgcgaggg catacccctc tccgtcatca gcctgcggct 720 caagcggccg gtcgatgcgg tctgcgccaa gctcgccgaa ctcggcatca cgcccaagct 780 cgagctttga ccggacgggc gccggggcat cgcgcccggg cgcccacccg ccggac 836 <210> 21 <211> 439 <212> DNA <213> Methylobacterium extorquens <400> 21 cgtgtgcaag tattccatgg ccacaccgga tgatgacagg gatgtccggc tatctggccc 60 ggcacgcaaa cccgtcaacg gggcgaccga tcaggctccc gcgcggccag tcagccagcg 120 gcggccaaac ggccgttcga ccggcggcgc ccgccggatt cgaccggact gccggtgaat 180 aacatccatt tgaaaaaagc ttctcttgtg gcaattgctt gatccaatta cgatcactat 240 cgcgtctcgc aagcctccg ttaaattata gtgacatggc attgaaaaat agaaggcatt 300 ccttcgcaac tgacttggcg tcgatgcata ttatagcatt taagagcttt tgtttcccga 360 aatggcaaca gctaagattc gccgcgtccc ggcctgcgat gatcgtcgaa tcgggcatgc 420 cgcgaccggg agacggcgc 439 <210> 22 <211> 42 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 22 gaccatgatt acgccaagct tatgaacacg gacgatttcg cc 42 <210> 23 <211> 28 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 23 cttgcacacg gtccggcggg tgggcgcc 28 <210> 24 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 24 cccgccggac cgtgtgcaag tattccatgg cc 32 <210> 25 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 25 aaaacgacgg ccagtgaatt cgcgccgtct cccggtcgc 39 <210> 26 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 26 aacccgcaga aacacgcttt 20 <210> 27 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 27 gaagctagct tatcgcgcca 20 <210> 28 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 28 tttctgcgcg taatctgctg 20 <210> 29 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 29 acgatgatgg cgagggtg 18 <210> 30 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 30 ggagcctgat gaacacggac 20 <210> 31 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 31 gcaatcatgc gccgtctc 18 <210> 32 <211> 580 <212> DNA <213> Methylobacterium extorquens <400> 32 gctggtatct gccgccgcgc tgccggcaga acctccggaa agacccattg tcgccccctg 60 tcgtttccgg cttgaccgtg gacgagattc gctcccgaga gcgtgaggga acgcacgcga 120 cctgtcgcgt ccggcgcgat cgccggatcg ggcgggatgc ggtgccgttc tgaagtgctg 180 tgagggagaa cattcgatgc gctcaggggc cgcttccgtc ttcttccccc tgttcgggct 240 cgtgctcacg gtcggcgcgg ccgatgcgca gacgccgccc gagccaggcg ctgcccggcc 300 gctggtgatc ccccaggtcg agggcgagcg ccagggcaag gtgctgggcc gggcgctcgc 360 ctgcggcgcg gagcgcgagc gggtcgatcg ggtgctgaga gccggccgcg agcgcatgat 420 ggcggcggtc ggccgggcgc tcacggagga gcgctacgcc ctggcgctcg acgacgcgat 480 gcgtctggag acgagcctgc ctgcgccctc ttcgatcgcg tgcgagaagg cgctggccgg 540 gctcgaacgc ctggagaagg cgccgtagac acccaaacga 580 <210> 33 <211> 1220 <212> DNA <213> Methylobacterium extorquens <400> 33 cggccctctc tccccggtac ggatgttgtg tggatgagcg cccatctggc gcagtccggc 60 cgggcggaaa gggtttttgc cgtttaccct tcgtcaacct taccgggtgc ttgctgccgg 120 caatgatgcg caccgcgtcc ctcaccgccg tcgatcccga agccctgctg ccgccgggcg 180 acgccgccct gcgcgcggcg atgcggggct ggcgcgaggc gctggcgcgg gagcggcgca 240 tggccgccaa cacggtcgag gcctacgagc gtgacctgcg ccagtttctg atccatcgcg 300 ccgcccgctc cggcacgccc accatcgccg ggctgatcgc cctgaagccc cgcgacctgc 360 gcgccttcat ggccgcgcgc cgggccgagg ggatcggtgg gcgctccctg atgcggatgc 420 tggcgggtct gcgctccttc gcccgcttcc tcgaacggga gggacacggc agcgtcgcgg 480 cgctcggcgc ggtgcgctcc cccaaggtcg agcgccgcct gccgcgcccg cttcccattt 540 ccgccgcgct ggcgatgacc gcgcccgaga cccgccccga cgacgaccgc gccccgtggg 600 tgctcgcccg cgacgcggcg gtgatcgccc tcctctacgg ctcgggcctg cgcatctcgg 660 aagccttagg gctcaccgcc cgcgacgcgc cgatgccggg catcgacgag gtgcgggtga 720 cgggcaaggg tgggaaggtc cgcgccgtgc cggtgctgcc ggcggtggcc gaggcggtgg 780 ccgcctacct gtcgctgtgc ccgcaccccc tcgatccgga ggggccgctc ttcgtcggcg 840 tgaagggcgg gccgctctcg ccgcgagtgg tccagtacgc ggtttccgcg ctccgcggcg 900 cgctcggcct gcccgagagc gcgaccccgc acgccctgcg acactccttc gcaacccatc 960 tgctcgcccg ccggggcgag ctgcgggcga tccaggaatt gctcggccac gcctcgctct 1020 cgaccacgca aatctacacc aaggtcgatg ccgcccgcct gatgagcgcc ttcgaggacg 1080 cccatccccg cgcccggcgg ctgccgccct cggaaccgcc gtcaccgcgt tccgaaaccg 1140 ttgatgcaga gcaaggcacg agcgcccgat ccggctatgc tgtccggcag gacaggccgg 1200 tggagcgtag aaatgcgtga 1220 <210> 34 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 34 gaccatgatt acgccaagct tgctggtatc tgccgccgc 39 <210> 35 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 35 gagagggccg tcgtttgggt gtctacggcg 30 <210> 36 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 36 acccaaacga cggccctctc tccccggtac 30 <210> 37 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 37 aaaacgacgg ccagtgaatt ctcacgcatt tctacgctcc acc 43 <210> 38 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 38 agatcgagac gagaagcg 18 <210> 39 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 39 ccacacaaca tccgtacc 18 <210> 40 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 40 gaccatgatt acgccaagc 19 <210> 41 <211> 18 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 41 ccacacaaca tccgtacc 18 <210> 42 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 42 cagaacctcc ggaaagaccc 20 <210> 43 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> artificial primer <400> 43 tccacacaac atccgtaccg 20

Claims (17)

Recombinant vector comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase. The method of claim 1, wherein the recombinant vector is a nucleic acid sequence encoding methylglyoxal synthase, a nucleic acid sequence encoding glycerol dehydrogenase, and methylglyoxal reductase or lact. The recombinant vector further comprises one or more selected from the group consisting of nucleic acid sequences encoding aldehyde reductase (lactaldehyde reductase). Methylotroph containing a nucleic acid sequence encoding lactoylglutathione lyase in an inactive state. The method of claim 3, wherein the methylotrophic bacteria are recombinant comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase. Transformed with a vector, methyl tropism. The method of claim 4, wherein the recombinant vector is a nucleic acid sequence encoding methylglyoxal synthase, a nucleic acid sequence encoding glycerol dehydrogenase, and methylglyoxal reductase or lactate. The methyltrophic bacteria additionally comprising at least one selected from the group consisting of a nucleic acid sequence encoding an aldehyde reductase (lactaldehyde reductase). The method of claim 3, wherein the methyltrophic bacteria are a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a methylglyoxal reductase or Lactaldehyde reductase (lactaldehyde reductase) will be transformed with a recombinant vector containing at least one selected from the group consisting of a nucleic acid sequence encoding a methyl bacterium. The method of claim 3, wherein the methylotrophic bacteria are Methylobacterium extorquens , Methylobacterium suomiense , Methylobacterium platani, Methylobacterium Ad. Hash boom ( Methylobacterium adhaesivum ), methylobacterium soli (Methylobacterium soli) and methylobacterium chloromethanicum (Methylobacterium chloromethanicum ) that is one or more selected from the group consisting of, methyltrophic bacteria. A composition for producing 1,2-propylene glycol (PDO) containing methylotroph containing a nucleic acid sequence encoding a lactoylglutathione lyase in an inactivated state. The method of claim 8, wherein the methylotrophic bacteria are recombinant comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase. The composition for the production of 1,2-PDO, which was transformed with a vector. The method of claim 9, wherein the recombinant vector is a nucleic acid sequence encoding methylglyoxal synthase, a nucleic acid sequence encoding glycerol dehydrogenase, and methylglyoxal reductase or lactate. The composition for producing 1,2-PDO, which additionally comprises at least one selected from the group consisting of a nucleic acid sequence encoding an aldehyde reductase (lactaldehyde reductase). The method of claim 8, wherein the methylotrophic bacteria are a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a methylglyoxal reductase or The composition for production of 1,2-PDO, which is transformed with a recombinant vector comprising at least one selected from the group consisting of nucleic acid sequences encoding lactaldehyde reductase. The method of claim 8, wherein the methylotrophic bacteria are Methylobacterium extorquens , Methylobacterium suomiense , Methylobacterium platani, Methylobacterium Ad. Hash boom ( Methylobacterium adhaesivum ), methylobacterium soli (Methylobacterium soli) and methylobacterium chloromethanicum (Methylobacterium chloromethanicum ) is one or more selected from the group consisting of, 1,2-PDO production composition. 1,2-propylene glycol (propanediol; PDO) comprising a culture step of culturing methylotroph containing a nucleic acid sequence encoding lactoylglutathione lyase in an inactivated state. Production method. The method of claim 13, wherein the methylotrophic bacteria are recombinant comprising at least one selected from the group consisting of a nucleic acid sequence encoding Formolase and a nucleic acid sequence encoding dihydroxyacetone kinase. Transformed with a vector, 1,2-PDO production method. The method of claim 14, wherein the recombinant vector comprises a nucleic acid sequence encoding methylglyoxal synthase, a nucleic acid sequence encoding glycerol dehydrogenase, and methylglyoxal reductase or lactate. The method for producing 1,2-PDO further comprises at least one selected from the group consisting of a nucleic acid sequence encoding an aldehyde reductase (lactaldehyde reductase). The method of claim 3, wherein the methyltrophic bacteria are a nucleic acid sequence encoding a methylglyoxal synthase, a nucleic acid sequence encoding a glycerol dehydrogenase, and a methylglyoxal reductase or The composition for production of 1,2-PDO, which is transformed with a recombinant vector comprising at least one selected from the group consisting of nucleic acid sequences encoding lactaldehyde reductase. The method of claim 13, wherein the methylotrophic bacteria are Methylobacterium extorquens , Methylobacterium suomiense , Methylobacterium platani, Methylobacterium Ad. Hash boom ( Methylobacterium adhaesivum ), methylobacterium soli (Methylobacterium soli) and methylobacterium chloromethanicum (Methylobacterium chloromethanicum ) is one or more selected from the group consisting of, the production method of 1,2-PDO.

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