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Method for producing PHA (polyhydroxyalkanoate) by using pseudomonas corrugata type II synthase

A technology of Pseudomonas and unicellular, which is applied in the field of producing PHA by using Pseudomonas wrinkle type II synthase, and can solve the problems of low production volume and high production cost of PHA

Pending Publication Date: 2021-06-04
LIAONING UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to solve the above problems, the present invention provides a method for Pseudomonas rugosa to construct type II chimeric enzymes to produce PHA, to solve the problems of low PHA production and high production costs

Method used

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  • Method for producing PHA (polyhydroxyalkanoate) by using pseudomonas corrugata type II synthase
  • Method for producing PHA (polyhydroxyalkanoate) by using pseudomonas corrugata type II synthase
  • Method for producing PHA (polyhydroxyalkanoate) by using pseudomonas corrugata type II synthase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1 uses Pseudomonas rugosa type II synthase to produce PHA with sodium oleate as carbon source

[0031] Strain: Pseudomonas rugosa pBBR1MCS2-phaC2

[0032] Medium:

[0033] 1. LB medium: 5g / l yeast extract, 10g / l tryptone, 10g / l NaCl, sterilized at 121°C for 15-20min after aliquoting.

[0034] 2. LB agar medium (pH 7.0-7.2): After adding the above ingredients, add 15g / l agar powder, subpackage and extinguish at 121°C for 15-20min.

[0035] 3. Mineral salt fermentation medium:

[0036] Table 1 Mineral salt fermentation medium component table

[0037]

[0038] Culture method: Culture Pseudomonas rugosa on LB agar medium at 30°C for about 12 hours, pick a single colony and inoculate it in 10ml of LB medium, shake at 30°C, 200rpm, 8-12h, as seed solution. Then add 1% sodium oleate to component II to prepare a mineral salt fermentation medium as a carbon source, inoculate the recombinant bacteria of pBBR1MCS2-phaC2 (preserved from our laboratory) with a 5% i...

Embodiment 2

[0062] Example 2 proves that Pseudomonas rugosa type II synthase produces PHA with sodium oleate as carbon source

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Abstract

The invention relates to the field of PHA biosynthesis, in particular to a novel PHA (polyhydroxyalkanoate) production method. A specific implementation scheme of the PHA production method comprises the following steps: taking pseudomonas corrugata as an original strain; verifying functional activity of pseudomonas corrugata pBBR1MCS2-phaC2 synthase through expression plasmids of pseudomonas corrugata pBBR1MCS2-phaC1 and pBBR1MCS2-phaC2 by using selected different carbon sources such as fructose, glycerol, lauric acid, sodium gluconate and sodium oleate; and confirming that the PHA can be generated by the recombinant strain by using the sodium oleate as the carbon source according to corresponding results. Therefore, successful construction of the chimeric enzyme in the future is ensured; and moreover, the universality of the substrate of the chimeric PHA synthase can be expanded by the chimeric PHA synthase, thereby increasing the yield of the PHA so as to greatly promote wide application of the PHA.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a method for producing PHA by using type II synthase of Pseudomonas rugosa. Background technique [0002] Hydroxy fatty acid ester (PHA) is an intracellular energy and carbon storage substance synthesized by many bacteria under nutrient imbalance conditions. PHA has good biocompatibility, biodegradability, hydrophobicity and unique properties brought by different functional groups. It is used as a tissue engineering material drug sustained release material. It is a biological material with good development potential, but At present, biosynthesized PHA has problems such as low yield, high cost, and single monomer component, which cannot meet the needs of new biomaterials in various fields. Researchers in academia and industry are trying to solve this key problem through various methods. technical problem. PHA synthesis requires the participation of many kinds of enz...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C12N1/20C12R1/38
CPCC12P7/625C12N1/20
Inventor 胡风庆赵小慧辛知纯
Owner LIAONING UNIVERSITY
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