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Recombinant microorganism having a producing ability of polylactate or its copolymers and method for preparing polylactate or its copolymers using the same

A technology of recombinant microorganisms and polylactate, which is applied in biochemical equipment and methods, botanical equipment and methods, recombinant DNA technology, etc., and can solve problems such as unavailability and difficulty in synthesizing PLA copolymers.

Inactive Publication Date: 2010-03-24
LG CHEM LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the synthesis of PLA by chemical synthesis of lactate, PLA homopolymers are readily available, but PLA copolymers composed of various types of monomers are difficult to synthesize and are not commercially available

Method used

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  • Recombinant microorganism having a producing ability of polylactate or its copolymers and method for preparing polylactate or its copolymers using the same
  • Recombinant microorganism having a producing ability of polylactate or its copolymers and method for preparing polylactate or its copolymers using the same
  • Recombinant microorganism having a producing ability of polylactate or its copolymers and method for preparing polylactate or its copolymers using the same

Examples

Experimental program
Comparison scheme
Effect test

preparation Embodiment 1

[0054] Preparation Example 1: Cloning of PHA Synthase Gene from Pseudomonas sp.6-19 and Construction of Table delivery vehicle

[0055] To isolate the PHA synthase gene (phaC1) derived from Pseudomonas sp.6-19 (KCTC 11027BP) used in the present invention Ps6-19 ), total DNA of Pseudomonas sp.6-19 was extracted. Based on phaC1 Ps6-19sequence (Ae-jin Song, Master's Thesis, Department of Chemical and Biomolecular Engineering, KAIST, 2004) to prepare primers (SEQ ID NO: 1 and SEQ ID NO: 2), and use the primers to perform polymerase chain reaction (PCR) , so as to get phaC1 Ps6-19 .

[0056] 5'-GAG AGA CAA TCA AAT CAT GAG TAA CAA GAG TAA CG-3' (SEQ ID NO: 1)

[0057] 5'-CAC TCA TGC AAG CGT CAC CGT TCG TGC ACG TAC-3' (SEQ ID NO: 2)

[0058] When the PCR product was analyzed by electrophoresis on an agarose gel, it was identified to correspond to phaC1 Ps6-19 1.7-kbp gene fragment of the gene. To express phaC1 Ps6-19 Synthase, a constitutive expression system of the opero...

preparation Embodiment 2

[0071] Preparative Example 2: Substrate-specific variants of PHA synthase from Pseudomonas sp.6-19 preparation of

[0072] Among the various PHA synthases, type II PHA synthases are known as medium-chain-length PHA (MCL-PHA) synthases for polymerizing substrates with a relatively large number of carbon atoms, and it is expected that MCL-PHA synthases can Applied to the production of PLA copolymer. Although the phaC1 synthase derived from Pseudomonas sp.61-3 is a type II PHA synthase, it differs from the phaC1 obtained according to the present invention Ps6-19 Synthases have a high degree of homology, but it has been reported that type II PHA synthases have a relatively wide range of substrate specificity (Matsusaki et al., J. Bacteriol., 180:6459, 1998), and have also been reported to be suitable for the production of Results of studies in mutations of short-chain-length PHA (SCL-PHA) (Takase et al., Biomacromolecules, 5:480, 2004). Based on the above studies, the presen...

preparation Embodiment 3

[0084] Preparation Example 3: Recombinant vector capable of expressing propionyl-CoA transferase from Clostridium propionicum build

[0085]To provide lactyl-CoA as a monomer required for the synthesis of PLA or PLA copolymers, propionyl-CoA transferase (cp-pct) from Clostridium propionicum was used. It is known that cp-pct is toxic to microorganisms. Typically, in an isopropyl-β-D-thiogalactopyranoside (IPTG)-inducible expression system using the tac or T7 promoter, which is widely used to express recombinant proteins, all recombinant Microorganisms die quickly. For this reason, it is considered suitable to produce PLA or PLA copolymers using a constitutive expression system in which cp-pct is expressed weakly but continuously as the microorganism grows. A fragment obtained by performing PCR on the chromosomal DNA of Clostridium propionicum using primers (SEQ ID NO: 17 and SEQ ID NO: 18) was used as cp-pct. In this regard, the NdeI site present in wild-type cp-pct was ...

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Abstract

Provided are a recombinant microorganism capable of producing polylactate (PLA) or hydroxyalkanoate-lactate copolymers and a method of preparing PLA or hydroxyalkanoate-lactate copolymers using the same. The recombinant microorganism has both a gene encoding a propionyl-CoA transferase from Megasphaera elsdenii and a gene encoding a polyhydroxyalkanoate (PHA) synthase using lactyl-CoA as a substrate. A propionyl-CoA transferase from Megasphaera elsdenii is introduced into the recombinant microorganism to effectively provide lactyl-CoA, thereby enabling efficient preparation of PLA or PLA copolymers.

Description

technical field [0001] The present invention relates to a recombinant microorganism and a method for producing PLA or a copolymer thereof by using the recombinant microorganism, the recombinant microorganism having both a gene encoding propionyl-CoA transferase from Megasphaera elsdenii and an encoding method for use Lactyl-CoA is a substrate for the polyhydroxyalkanoate (PHA) synthase gene and is capable of producing polylactate (PLA) or hydroxyalkanoate-lactate copolymers. Background technique [0002] Polylactate (PLA) is a common biodegradable polymer derived from lactate, which is widely used in commercial and biomedical fields. Currently, although the preparation of PLA involves the polymerization of lactic acid esters produced by microbial fermentation, only PLA with a low molecular weight of about 1000˜5000 Daltons is produced through the direct polymerization of lactic acid esters. In order to synthesize PLA with a molecular weight above 100,000 Daltons, PLA with a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31
CPCC12N9/10C12N9/1029C12P7/625C12N15/11C12N5/14C12N1/205A01H1/06
Inventor 朴时载梁宅镐李相贤李恩政姜惠玉金泰完
Owner LG CHEM LTD
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