WO2012169819A2 - Procédé de préparation de polyhydroxyalcanoate contenant du 2-hydroxybutyrate en tant que monomère - Google Patents

Procédé de préparation de polyhydroxyalcanoate contenant du 2-hydroxybutyrate en tant que monomère Download PDF

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WO2012169819A2
WO2012169819A2 PCT/KR2012/004523 KR2012004523W WO2012169819A2 WO 2012169819 A2 WO2012169819 A2 WO 2012169819A2 KR 2012004523 W KR2012004523 W KR 2012004523W WO 2012169819 A2 WO2012169819 A2 WO 2012169819A2
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gene encoding
hydroxybutyrate
amino acid
acid sequence
coa
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WO2012169819A3 (fr
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이상엽
박시재
이승환
송봉근
이태우
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Korea Research Institute of Chemical Technology KRICT
Korea Advanced Institute of Science and Technology KAIST
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Korea Advanced Institute of Science and Technology KAIST
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    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
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    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C12P7/625—Polyesters of hydroxy carboxylic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters

Definitions

  • the present invention relates to a method for producing polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer, and more specifically, to the polyhydroxyalkanoate containing 2-hydroxybutyrate using metabolic engineering. It relates to a method for preparing from a recombinant strain.
  • PHAs Polyhydroxyalkanoates
  • PHA synthase which uses several kinds of hydroxyacyl-CoA as a substrate.
  • hydroxyacyl-CoA has an asymmetric center at the carbon position of the hydroxy group, it is all an (R) -configuration.
  • various hydroxyacyl-CoAs such as 3-hydroxyacyl-CoAs, 4-hydroxyacyl-CoAs, 5-hydroxyacyl-CoAs, 6-hydroxyacyl-CoAs, and the like, PHA synthase is the most preferred substrate.
  • Roxiasil-CoAs (3HA-CoAs).
  • PLA polylactic acid
  • (D) -Lactyl-CoA is propionyl-CoA propionyl of Clostridium propionicum which converts (D) -lactate to (D) lactyl-CoA using acetyl-CoA as a CoA donor in the recombinant E. coli. CoA transferase) was employed to enable production.
  • lactate polymer (PLA) with lactyl-CoA Pseudomonas sp. was genetically engineered to use lactyl-CoA as a substrate. 6-19 PHA synthase (PhaC1 ps6-19 ) was employed.
  • the present inventors earnestly endeavored to develop a method of producing 2-hydroxybutyl-CoA, which is a substrate of PhaC1 ps6-19 , using propionyl-CoA, in order to develop PHA synthesized by a new route, propionate propionyl.
  • Propionyl-CoA synthetase (PrpE) from Ralstonia eutropha converting to -CoA, pyruvate formate lyase (PflB) converting propionyl-CoA to 2-ketobutyrate and 2-hydroxybutyrate dehydrogenase converting 2-ketobutyrate to 2HB
  • PflB pyruvate formate lyase
  • An object of the present invention is to provide a method for producing polyhydroxyalkanoate containing 2-hydroxybutyrate, a biodegradable polymer as a monomer, and a recombinant microorganism producing the polymer.
  • the present invention is a microorganism having a pathway for biosynthesis of acetyl-CoA from a carbon source, the gene encoding the lacdate dehydrogenase is deleted, and the polyhydroxy alkanoate synthase is encoded
  • a gene encoding the propionyl-CoA transferase, a gene encoding the D-2 hydroxy acid dehydrogenase, a gene encoding the propionyl-CoA synthetase, and a gene encoding the pyruvate formate lyase Provided is a recombinant microorganism having a polyhydroxyalkanoate producing ability containing 2-hydroxybutyrate as a monomer.
  • the present invention also relates to a microorganism having a pathway for biosynthesis of acetyl-CoA from a carbon source, wherein the gene encoding lacdate dehydrogenase is deleted, and the gene encoding polyhydroxy alkanoate synthase and propionyl- 2-hydroxy, characterized by introducing a gene encoding CoA transferase, a gene encoding D-2 hydroxy acid dehydrogenase, a gene encoding propionyl-CoA synthetase, and a gene encoding pyruvate formate lyase
  • a method for producing a recombinant microorganism having a polyhydroxyalkanoate-generating ability containing butyrate as a monomer is provided.
  • the present invention also comprises the steps of culturing the recombinant microorganism in a medium containing glucose and propionic acid to produce polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer; And it provides a method for producing a polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer comprising the step of obtaining a polyhydroxyalkanoate containing the produced 2-hydroxybutyrate as a monomer. do.
  • 1 shows a biosynthetic pathway for synthesizing polyhydroxyalkanoate containing 2-hydroxybutyrate as monomer from glucose, propionic acid and 3HB.
  • the present invention is a microorganism having a pathway for synthesizing acetyl-CoA from a carbon source, the gene encoding the lacdate dehydrogenase is deleted, the gene encoding a polyhydroxy alkanoate synthase and A gene encoding propionyl-CoA transferase, a gene encoding D-2 hydroxy acid dehydrogenase, a gene encoding propionyl-CoA synthetase, and a gene encoding pyruvate formate lyase are introduced.
  • the present invention relates to a recombinant microorganism having a polyhydroxyalkanoate producing ability containing oxybutyrate as a monomer.
  • the propionyl -CoA transferase may be characterized in that the mutant enzyme of the transferase or propionyl propionyl -CoA -CoA transferase of C. propionicum of C. propionicum Pct540.
  • the polyhydroxy alkanoate synthetase is Pseudomonas sp. It may be characterized by the 6-19 PHA synthase or its mutant enzyme.
  • the mutant of the PHA synthase is one or more mutations selected from the group consisting of E130D, S325T, L412M, S477R, S477H, S477F, S477Y, S477G, Q481M, Q481K and Q481R in the amino acid sequence of SEQ ID NO: 14 It may be characterized in that it has an amino acid sequence comprising the amino acid sequence of E130D, S325T, L412M, S477G and Q481M mutation in the amino acid sequence of SEQ ID NO: 14 (C1335); Amino acid sequence (C1310) in which E130D, S477F and Q481K are mutated in the amino acid sequence of SEQ ID NO: 14; And in the amino acid sequence of SEQ ID NO: 14 E130D, S477F and Q481R may be characterized by having any one amino acid sequence selected from the group consisting of mutated amino acid sequence (C1312).
  • the recombinant microorganism may be introduced with a gene encoding ⁇ -keto thiolase and a gene encoding acetoacetyl-CoA reductase, gene encoding the ⁇ -keto thiolase Is phaA derived from Ralstonia eutropha, and the gene encoding acetoacetyl-CoA reductase may be phaB derived from Ralstonia eutropha .
  • the gene encoding the introduced ⁇ -ketothiolase and the gene encoding acetoacetyl-CoA reductase are expressed in recombinant microorganisms to convert acetyl-CoA to acetoacetyl-CoA and convert the acetoacetyl- Recombinant microorganisms in which CoA is converted to 3-hydroxybutyryl-CoA, wherein phaA and phaB are additionally introduced, can be extracted from glucose and propionic acid without the addition of 3-hydroxybutyrate from p (2HB-co -3HB) can be produced.
  • the mutated PhaC1 ps6-19 has various substrate specificities, lactate also because it is a type of 2-hydroxy acid, the 2HB-CoA by PhaC1 ps6-19 may be copolymerized in 2HB-containing polymer.
  • 3HB was chosen as the second monomer because 3-HB-CoA acts as an initiator in the synthesis of lactate containing polymers.
  • the variation PhaC1 ps6-19 and mutated Pct cp has a V193A mutation and four silent mutations of the nucleotide sequence (T78C, T669C, A1125G and T1158C) and Pct540 PhaC1 ps6-19d
  • Recombinant Escherichia coli containing PhaC1437 containing four variants of E130D, S325T, S477G and Q481K in E. coli efficiently produced P (3HB-co-LA) copolymer with high lactate fraction.
  • these enzymes were used for the biosynthesis of polyhydroxyalkanoates containing 2-hydroxybutyrate as monomer.
  • the gene encoding the D-2 hydroxy acid dehydrogenase is Lactobacillus lactis Origin panE It may be characterized in that, the gene encoding the propionyl-coA synthetase is Ralstonia eutropha Origin prpE It can be characterized as that the pyruvate formate lyase Clostridium difficile Origin pflB Can be characterized as being.
  • panE derived from Lactobacillus lactis was used as a gene encoding D-2 hydroxy acid dehydrogenase, but it was limited as long as it was expressed in the introduced host cell to express the activity of D-2 hydroxy acid dehydrogenase. Can be used without
  • a prpE derived from Ralstonia eutropha was used as a gene encoding a propionyl-CoA synthetase, it can be used without limitation as long as it is expressed in the introduced host cell and shows the activity of pyruvate formate lyase.
  • Ralstonia converts propionate to propionyl-CoA in recombinant E. coli metabolically engineered to express propionyl-CoA transferase gene.
  • the gene encoding the eutropha- derived propionyl-CoA synthetase (PrpE) was expressed, and the enzyme encoding pyruvate formate lyase (PflB), which converts propionyl-CoA to 2-ketobutyrate, and 2-ketobutyrate to 2HB.
  • PrpE eutropha- derived propionyl-CoA synthetase
  • PflB pyruvate formate lyase
  • the gene encoding 2-hydroxybutyrate dehydrogenase was expressed.
  • the introduction of phaA from Ralstonia eutropha, which is a gene encoding ⁇ -ketothiolase, and phaB, which is a gene encoding acetoacetyl-CoA reductase, to the recombinant microorganism of the present invention is pCnCAB.
  • phaA from Ralstonia eutropha which is a gene encoding ⁇ -ketothiolase
  • phaB which is a gene encoding acetoacetyl-CoA reductase
  • the present invention relates to a gene and a propie, which are deleted from a gene encoding lacdate dehydrogenase, and a polyhydroxy alkanoate synthase, in a microorganism having a pathway for biosynthesis of acetyl-CoA from a carbon source.
  • 2- characterized by introducing a gene encoding onyl-CoA transferase, a gene encoding D-2 hydroxy acid dehydrogenase, a gene encoding propionyl-CoA synthetase, and a gene encoding pyruvate formate lyase.
  • the present invention relates to a method for producing a recombinant microorganism having a polyhydroxyalkanoate producing ability containing hydroxybutyrate as a monomer.
  • the present invention comprises the steps of culturing the recombinant microorganism in a medium containing glucose and propionic acid to produce a polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer; And a process for producing polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer, comprising obtaining polyhydroxyalkanoate containing the produced 2-hydroxybutyrate as a monomer.
  • the polyhydroxyalkanoate containing 2-hydroxybutyrate as a monomer may be 2-hydroxybutyrate-co-3-hydroxybutyrate-co-lactate.
  • the medium may be characterized as further containing 3-hydroxybutyrate.
  • the E. coli XLdh strain from which the ldhA gene was removed from the chromosome of E. coli XL1-Blue was used as a host cell for P (2HB-co-3HB) copolymer synthesis.
  • the present inventors adopted a route for producing 2-ketobutyrate from propionyl-CoA to produce the 2-ketobutyrate.
  • a gene encoding the pyruvate formate lyase of Clostridium difficile and a gene encoding the propionyl-CoA synthetase of R. eutropha in a host cell through gene recombination. was expressed.
  • D-2-hydroxy acid dehydrogenases have selectivity for the optical isomers and use NADH as a cofactor to reduce 2-keto acid to D-2-hydroxy acid.
  • the D-2-hydroxy acid dehydrogenase is 2-ketoisocaproate, 2-ketoisovalerate, 2-ketovalorate, 2-ketobutyrate and It has a wide range of substrate specificities, including mandelate.
  • L. lactissub sp. Among various D-2-hydroxy acid dehydrogenases . 2HB was synthesized using D-2 hydroxy acid dehydrogenase, which is encoded by the panE gene of lactis Il1403.
  • 2HB when synthesizing p (2HB-co-3HB) polymer using recombinant E. coli, 2HB must be produced intracellularly from glucose, and 2HB is converted to 2HB-CoA by Pct540. do. In order to produce 3HB-CoA in cells, 3HB had to be added to the medium, and as the concentration of 3HB in the medium increased, the 2HB monomer fraction in the synthesized polymer decreased.
  • the present invention relates to a (2-hydroxybutyrate-co-3-hydroxybutyrate) polymer.
  • vector refers to a DNA preparation containing a DNA sequence operably linked to a suitable regulatory sequence capable of expressing DNA in a suitable host.
  • the vector may be a plasmid, phage particles, or simply a potential genomic insert. Once transformed into the appropriate host, the vector can replicate and function independently of the host genome, or in some cases can be integrated into the genome itself. Since plasmids are the most commonly used form of current vectors, “plasmid” and “vector” are sometimes used interchangeably in the context of the present invention. For the purposes of the present invention, it is preferred to use plasmid vectors.
  • Typical plasmid vectors that can be used for this purpose include (a) a replication initiation point that allows for efficient replication to include hundreds of plasmid vectors per host cell, and (b) host cells transformed with the plasmid vector. It has a structure comprising an antibiotic resistance gene and (c) a restriction enzyme cleavage site into which foreign DNA fragments can be inserted. Although no appropriate restriction enzyme cleavage site is present, the use of synthetic oligonucleotide adapters or linkers according to conventional methods facilitates ligation of the vector and foreign DNA.
  • the vector should be transformed into the appropriate host cell.
  • preferred host cells are prokaryotic cells.
  • Suitable prokaryotic host cells include E. coli DH5 ⁇ , E. coli JM101, E. coli K12, E. coli W3110, E. coli X1776, E. coli XL-1Blue (Stratagene), E. coli B, E. coli B21, etc. It includes.
  • E. coli strains such as FMB101, NM522, NM538 and NM539 and other prokaryotic species and genera may also be used. In addition to the aforementioned E.
  • strains of the genus Agrobacterium such as Agrobacterium A4, bacilli , such as Bacillus subtilis , Salmonella typhimurium or Serratia marghesen another variety of enteric bacteria and Pseudomonas (Pseudomonas) in strains such as marcescens) may be used as host cells.
  • an expression vector known in the art may be used, and it is preferable to use a pET family vector (Novagen).
  • a pET family vector Novagen
  • histidine groups are bound to the ends of the expressed protein, and thus the protein can be effectively purified.
  • a general method known in the art may be used, and specifically, it may be separated by a chromatographic method using Ni-NTA His-binding resin (Novagen).
  • the recombinant vector may be characterized in that the pET-SLTI66, the host cell may be characterized in that E. coli or Agrobacterium.
  • control sequence refers to a DNA sequence essential for the expression of a coding sequence operably linked in a particular host organism.
  • regulatory sequences include promoters for performing transcription, any operator sequence for regulating such transcription, sequences encoding suitable mRNA ribosomal binding sites, and sequences that control the termination of transcription and translation.
  • suitable control sequences for prokaryotes include promoters, optionally operator sequences, and ribosomal binding sites.
  • Eukaryotic cells include promoters, polyadenylation signals, and enhancers. The factor that most influences the amount of gene expression in the plasmid is the promoter.
  • an SR ⁇ promoter As the promoter for high expression, an SR ⁇ promoter, a promoter derived from cytomegalovirus, and the like are preferably used.
  • any of a wide variety of expression control sequences can be used in the vector.
  • Useful expression control sequences include, for example, early and late promoters of SV40 or adenovirus, lac system, trp system, TAC or TRC system, T3 and T7 promoters, major operator and promoter regions of phage lambda, fd code protein Regulatory region of, promoter for 3-phosphoglycerate kinase or other glycolysis enzymes, promoters of the phosphatase such as Pho5, promoter of yeast alpha-crossing system and gene expression of prokaryotic or eukaryotic cells or viruses Other sequences known to modulate and various combinations thereof.
  • the T7 promoter can be usefully used to express the proteins of the invention in E. coli.
  • Nucleic acids are "operably linked” when placed in a functional relationship with other nucleic acid sequences. This may be genes and regulatory sequence (s) linked in such a way as to allow gene expression when appropriate molecules (eg, transcriptional activating proteins) bind to regulatory sequence (s).
  • the DNA for a pre-sequence or secretion leader is operably linked to the DNA for the polypeptide when expressed as a shear protein that participates in the secretion of the polypeptide;
  • a promoter or enhancer is operably linked to a coding sequence when it affects the transcription of the sequence;
  • the ribosomal binding site is operably linked to a coding sequence when it affects the transcription of the sequence;
  • the ribosomal binding site is operably linked to a coding sequence when positioned to facilitate translation.
  • "operably linked” means that the linked DNA sequence is in contact, and in the case of a secretory leader, is in contact and present within the reading frame.
  • enhancers do not need to touch. Linking of these sequences is performed by ligation (linking) at convenient restriction enzyme sites. If such sites do not exist, synthetic oligonucleotide adapters or linkers according to conventional methods are used.
  • expression vector generally refers to a fragment of DNA that is generally double stranded as a recombinant carrier into which fragments of heterologous DNA have been inserted.
  • heterologous DNA refers to heterologous DNA, which is DNA not naturally found in host cells.
  • the gene must be operably linked to transcriptional and translational expression control sequences that function in the selected expression host.
  • the expression control sequence and the gene of interest are included in one expression vector including the bacterial selection marker and the replication origin. If the host cell is a eukaryotic cell, the expression vector must further comprise an expression marker useful in the eukaryotic expression host.
  • Host cells transformed or transfected with the expression vectors described above constitute another aspect of the present invention.
  • transformation means introducing DNA into a host so that the DNA is replicable as an extrachromosomal factor or by chromosomal integration.
  • transfection means that the expression vector is accepted by the host cell whether or not any coding sequence is actually expressed.
  • the relative strength of the sequence, the controllability, and the compatibility with the DNA sequences of the present invention should be considered, particularly with regard to possible secondary structures.
  • Single cell hosts may be selected from a host for the selected vector, the toxicity of the product encoded by the DNA sequence of the invention, the secretory properties, the ability to accurately fold the protein, culture and fermentation requirements, the product encoded by the DNA sequence of the invention from the host. It should be selected in consideration of factors such as the ease of purification.
  • one skilled in the art can select a variety of vector / expression control sequence / host combinations capable of expressing the DNA sequences of the invention in fermentation or large scale animal culture.
  • a binding method binding method
  • a panning method a panning method
  • a film emulsion method and the like can be applied.
  • Table 1 shows the strains and plasmids used in the examples below.
  • Example 1 (2-hydroxybutyrate-co-3-hydroxybutyrate-co-lactate) polymer from glucose, propionic acid and 3HB using recombinant strain [P (2HB-co-3HB-co-LA) Biosynthesis
  • E. coli XL1-Blue (Stratgene Cloning System, USA) was used as a host cell for gene cloning, and cultured in 37 ° C. LB medium.
  • E. coli XLdh strain was incubated for 72 h at 250 rpm at 30 ° C using MR medium containing 20 g / L glucose, propionic acid and the desired concentration of 3HB, and the concentrations of 2HB, 3HB and LA depending on the culture conditions. Adjusted.
  • Plasmid p619C1437-pct540 under the PHA synthase operon promoter of Ralstonia eutropha , contains four variants of E130D, S325T, S477G and Q481K in PHA synthase of Pseudomonas sp.6-19 and propionyl-CoA transferase of C. propionicum .
  • PanE gene of lactis Il1403 was introduced and shaken cultured for 30 hours at 30 ° C. and 250 rpm using MR medium containing 20 g / L glucose and the desired concentration of propionate and 3HB, depending on the culture conditions of 2HB and 3HB. The concentration was adjusted to prepare p (2HB-co-3HB-co-LA).
  • the primers used in the preparation of the recombinant plasmid in this example are shown in Table 2.
  • the expression vector pZE12-MCS (EXPRESSYS, USA) is a PHA biosynthetic gene transcription terminator of muti-cloning sites (MCS) and Ralstonia eutropha of pTacLac (Lee et al . Appl. Microbiol. Biotechnol. 79,633-641,2008) . Transformed to have.
  • the pTacLac vector is a pTrc99A (SspI digested gene fragment containing a Tac promoter and transcription terminator obtained by cleaving the pTac99A (Park and Lee, J. Bacteriol. 185, 5391-5397, 2003) vector with SspI). Pharmacia Biotech, Sweden), and pTac99A was prepared by replacing the trc promoter of pTrc99A (Pharmacia Biotech, Sweden) with a tac promoter obtained by cleaving pKK223-3 (Pharmacia Biotech, Sweden) with PvuII and EcoRI.
  • a PHA biosynthetic gene transcription terminator gene fragment was obtained by PCR using the primers of SEQ ID NOS: 1 to 2 from p619C1437-pct540, and the obtained gene fragment was used as a template. PCR was performed using primers of SEQ ID NO: 2 and SEQ ID NO: 3 to obtain PHA biosynthetic gene transcription terminators of RBS, pTacLac MCS and R. eutropha . The PCR product was digested with MfeI and AvrII and inserted into pZE12-MCS digested with EcoRI / AvrII to prepare pKE12-MCS.
  • the pKA32-MCS was PCR using primers SEQ ID NOs: 6 to 7 to amplify a gene fragment including a P LlacO-1 promoter, MCS, and a PHA biosynthetic gene transcription terminator of R. eutropha , and amplified the gene fragment.
  • pBBR1MCS2 ⁇ GenBank no. U23751; Kovach et al .
  • PKM22-MCS was prepared by insertion into the SspI site of Gene , 166,175-176,1995).
  • pKM22-MCS was inserted with lpanE gene amplified by PCR using primers of SEQ ID NOs: 8-9, pKM22PanE was prepared, and chromosomal DNA of C. difficile in pKM22PanE, In 630 hadA and C. difficile chromosomal DNA amplified by PCR using the primers of SEQ ID NOS: 10-11, pKM22PanEHadAPflB was prepared by inserting the pflB gene amplified by PCR using primers SEQ ID NOs: 12-13.
  • the hadA gene is an enzyme that converts 2-hydroxyisovalerate and 2-hydroxyisocaproate into 2-hydroxyisovaleryl-CoA and 2-hydroxyisocaproyl-CoA, but the role of making 2HB-CoA in the present invention was much lower than that of pct540.
  • Recombinant E. coli XLdh transformed with p619C1437-pct540, pKA32-RePrpE, pKM22PanEHadAPflB plasmids, expressing the phaC437, pct540, prpE, pflB and panE genes is glucose, at least 6.3% by weight of P (73 mol%) from propionic acid, 3HB 2HB-co-27 mol% LA) polymer was prepared (Table 3).
  • the present invention provides a recombinant microorganism capable of producing a polyhydroxyalkanoate containing 2-hydroxybutyrate, a new biodegradable polymer as a monomer, and a 2-hydroxybutyrate monomer, characterized by culturing the recombinant microorganism. There is an effect of providing a method for producing a polyhydroxyalkanoate containing.

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Abstract

La présente invention concerne un procédé de préparation de polyhydroxyalcanoate contenant du 2-hydroxybutyrate en tant que monomère et, plus précisément, un procédé de préparation, à partir de souches recombinantes et par génie métabolique, de polyhydroxyalcanoate contenant du 2-hydroxybutyrate. La présente invention concerne un microorganisme recombinant capable de générer du polyhydroxyalcanoate contenant du 2-hydroxybutyrate, un polymère biodégradable inédit, en tant que monomère, ainsi qu'un procédé de préparation de polyhydroxyalcanoate contenant du 2-hydroxybutyrate en tant que monomère, impliquant la culture dudit microorganisme recombinant.
PCT/KR2012/004523 2011-06-08 2012-06-08 Procédé de préparation de polyhydroxyalcanoate contenant du 2-hydroxybutyrate en tant que monomère Ceased WO2012169819A2 (fr)

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KR1020110055113A KR101273599B1 (ko) 2011-06-08 2011-06-08 2-하이드록시부티레이트를 모노머로 함유하고 있는 폴리하이드록시알카노에이트의 제조방법

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WO2017171260A1 (fr) * 2016-03-28 2017-10-05 주식회사 엘지화학 Biopolymère liquide, son utilisation et procédé de préparation
CN110382699A (zh) * 2017-02-28 2019-10-25 韩国科学技术院 使用2-羟基异己酸-CoA转移酶生成聚羟基链烷酸酯的方法
JP2020508665A (ja) * 2017-02-28 2020-03-26 コリア アドバンスト インスティチュート オブ サイエンス アンド テクノロジー ２−ヒドロキシイソカプロエート−ＣｏＡ転移酵素を用いたポリヒドロキシアルカノエートの製造方法
EP3594350A4 (fr) * 2017-02-28 2020-12-09 Korea Advanced Institute of Science and Technology Procédé de production de polyhydroxyalcanoate à l'aide de 2-hydroxyisocaproate-coa transférase
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CN110382699B (zh) * 2017-02-28 2024-04-05 韩国科学技术院 使用2-羟基异己酸-CoA转移酶生成聚羟基链烷酸酯的方法
CN114561333A (zh) * 2022-02-23 2022-05-31 山东第一医科大学(山东省医学科学院) 一种转化支链氨基酸的工程菌及在制备治疗枫糖尿症产品中的应用
CN114561333B (zh) * 2022-02-23 2023-08-25 山东第一医科大学(山东省医学科学院) 一种转化支链氨基酸的工程菌及在制备治疗枫糖尿症产品中的应用
CN115851510A (zh) * 2022-10-13 2023-03-28 深圳中科翎碳生物科技有限公司 盐单胞菌及其在联产四氢嘧啶和聚羟基脂肪酸酯的应用
CN115851510B (zh) * 2022-10-13 2023-09-12 深圳中科翎碳生物科技有限公司 盐单胞菌及其在联产四氢嘧啶和聚羟基脂肪酸酯的应用

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