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Copolymer containing 3-hydroxyalkanoate unit and lactate unit, and its manufacturing method

A technology of lactate terpolymer and hydroxyalkanoate, applied in the field of copolymers, can solve the problems of weak PHA synthase activity, difficult to remove solvent or chain coupling agent, complicated preparation method of high molecular weight PLA, etc.

Active Publication Date: 2009-12-30
LG CHEM LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has the following disadvantages: as the preparation method of high molecular weight PLA is complicated due to the addition of solvent or chain coupling agent, and it is not easy to remove the solvent or chain coupling agent
So far, it has been reported that the PHA synthase activity to lactyl-CoA has been detected in vitro, but the PHA synthase activity to lactyl-CoA is very weak (Zhang et al., Appl. Microbiol. Biotechnol., 56: 131, 2001 ; Valentinand Steinbuchel, Appl. Microbiol. Biotechnol., 40:699, 1994)

Method used

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  • Copolymer containing 3-hydroxyalkanoate unit and lactate unit, and its manufacturing method
  • Copolymer containing 3-hydroxyalkanoate unit and lactate unit, and its manufacturing method
  • Copolymer containing 3-hydroxyalkanoate unit and lactate unit, and its manufacturing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Embodiment 1 comprises the construction of the recombinant plasmid of pct gene and PHA synthase gene

[0040] Recombinant plasmids containing pct gene and PHA synthase gene, pPs619C1300-CPPCT and pTacCpPctNCvEC, were constructed to prepare copolymers containing 3-hydroxyalkanoate units and lactate units.

[0041] (1) Construction of pPs619C1300-CPPCT plasmid The propionyl-CoA transferase (CP-PCT) gene derived from Clostridium propionicum was used as the pct gene, and the PHA derived from Pseudomonas 6-19 was used to synthesize Enzyme gene as PHA synthase gene.

[0042] Such as figure 1 Construct the operon of the constitutive expression system that simultaneously expresses PHA synthase and CP-PCT. CP-PCT is known to be toxic to host microorganisms. That is, in the tac promoter or T7 promoter expression system induced with IPTG (this system is widely used for the expression of recombinant proteins), all the microorganisms died shortly after adding the inducer. For th...

Embodiment 2

[0075] Example 2: Preparation of MCL 3-hydroxyalkanoate-lactate copolymer

[0076] Transform Escherichia coli WB101 (Park and Lee, J.Bacteriol.185, 5391-5397, 2003) with the recombinant plasmid pPs619C1300-CPPCT comprising pct gene and PHA synthase gene constructed in Example 1, to obtain Escherichia coli WB101 / pPs619C1300-CPPCT. WB101 is reported to be a fadB E. coli mutant effective in producing MCL-PHA (Korean Patent No. 10-0447531).

[0077] Transformants were cultivated in the following two steps to obtain MCL 3-hydroxyalkanoate-lactate copolymers: first, in 100 mL LB medium containing 100 mg / L ampicillin (Bacto TM Tryptone (BD) 10g / L, Bacto TM The transformed recombinant Escherichia coli WB101 / pPs619C1300-CPPCT was cultured in yeast extract (BD) 5g / L, NaCl(amresco) 10g / L) for 24 hours, and then the culture medium was centrifuged at 1000g for 15 minutes at 4°C to collect the cells .

[0078] In the LB medium (Bacto TM Tryptone (BD) 10g / L, Bacto TM The collected cel...

Embodiment 3

[0081] Example 3: Preparation of 3-hydroxybutyrate-MCL 3-hydroxyalkanoate-lactate terpolymer

[0082] Transform Escherichia coli WB101 [W3110(fadB::Km) Park and Lee, J.Bacteriol.185:5391,2003] with the recombinant plasmid pPs619C1300-CPPCT comprising pct gene and PHA synthase gene constructed in Example 1, To obtain Escherichia coli WB101 / pPs619C1300-CPPCT. WB101 is reported to be a fadB E. coli mutant effective in producing MCL-PHA (Korean Patent No. 10-0447531).

[0083] The transformants were cultivated to obtain 3-hydroxybutyrate-MCL 3-hydroxyalkanoate-lactate terpolymers by the following two steps: first, in 100 mL LB medium containing 100 mg / L ampicillin ( Bacto TM Tryptone (BD) 10g / L, Bacto TM The transformed recombinant Escherichia coli WB101 / pPs619C1300-CPPCT was cultured in yeast extract (BD) 5g / L, NaCl(amresco) 10g / L) for 24 hours, and then the culture medium was centrifuged at 1000g for 15 minutes at 4°C to collect the cells .

[0084] In the LB medium (Bacto ...

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Abstract

The present invention relates to a copolymer comprising 3-hydroxyalkanoate monomer unit and lactate monomer unit, or their preparing method. More specifically, the present invention relates to a method for preparing a copolymer comprising lactate monomer and 3-hydroxyalkanoate monomer, wherein the method comprises culturing a cell or plant comprising the gene of enzyme converting lactate and 3-hydroxyalkanoate into lactyl-CoA and 3-hydroxyalkanoyl-CoA, respectively, and polyhydroxyalkanoate synthase gene together, and the copolymer made by the method. The copolymer of the present invention is a biodegradable polymer being able to be usefully used instead of conventional synthetic plastic, and the copolymer can be used also for medical use.

Description

technical field [0001] The present invention relates to a copolymer comprising 3-hydroxyalkanoate monomer units and lactate monomer units, and a method for preparing such a polymer. Background technique [0002] Polylactate (PLA) is a typical biodegradable polymer derived from lactate, which has various uses as a conventional or medical polymer. Currently, PLA is produced by polymerizing lactate produced by fermenting microorganisms, however, only low molecular weight (1000-5000 Dalton) PLA can be produced by direct polymerizing lactate. For the synthesis of high molecular weight (>100,000 Daltons) PLA, a method of polymerizing low molecular weight PLA obtained by direct polymerization of lactate via a chain coupling agent can be used. However, it has disadvantages such as complicating the preparation method of high-molecular-weight PLA due to the addition of a solvent or a chain coupling agent, and also not being easy to remove the solvent or chain coupling agent. Curr...

Claims

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Application Information

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IPC IPC(8): C08G63/91C08G63/08
CPCC08G63/00C08G63/06C08G63/08C08G63/78C08L67/00C08L2203/02C12N15/52C12P7/56
Inventor 朴时载李相贤李恩政姜惠玉金泰完梁宅镐李相烨
Owner LG CHEM LTD
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