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Method for extracting and purifying polyhydroxyalkanoate

A technology of polyhydroxyalkanoate and microbial bacteria, which is applied in the downstream post-processing field of bioengineering, can solve the problems of complicated extraction steps, high cost of chemical substances, difficult downstream sewage treatment, etc., achieve simple and safe production environment, reduce extraction cost, The effect of reducing processing difficulty

Active Publication Date: 2020-04-17
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] The present inventors found unexpectedly, by treating the microbial thalline that produces polyhydroxyalkanoate (PHA) with hot water alone, especially superheated water, can obtain thalline cell wall broken lysate, and avoided organic solvent and / or The use of surfactants overcomes the problems of complicated traditional PHA extraction steps, high cost of adding chemical substances, and difficult treatment of downstream sewage.

Method used

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  • Method for extracting and purifying polyhydroxyalkanoate
  • Method for extracting and purifying polyhydroxyalkanoate
  • Method for extracting and purifying polyhydroxyalkanoate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: Superheated water method extracts and purifies PHA from the fermented liquid containing PHA thalline

[0050] method of extraction:

[0051] (1) Centrifugation and resuspension of bacteria for desalting

[0052] Halomonas containing 60% PHA (P3HB, P34HB can be) 1 250mL of the fermentation broth of the bacteria was placed symmetrically in a high-speed centrifuge after balancing, and the centrifugation conditions were: speed 6000rpm, time 20min. After the centrifugation, carefully remove the supernatant, pay attention not to lose the precipitate as much as possible, then add water to restore the volume, stir well and continue centrifuging, the centrifugation conditions are: 6000rpm, time 20min. After centrifugation, discard the supernatant, restore the volume of the bacteria to 250mL, stir evenly, pour the washed bacteria solution into a 500mL beaker, and prepare to break the wall.

[0053] (2) Cell wall breaking and PHA release of bacterial liquid

[00...

Embodiment 2

[0061] Embodiment 2: Superheated water protease method extracts and purifies PHA from the fermented liquid containing PHA thallus

[0062] method of extraction:

[0063] (1) Centrifugation and resuspension of bacteria for desalting

[0064] Halomonas containing 60% PHA (P3HB, P34HB can be) 1 250mL of the fermentation broth of the bacteria was placed symmetrically in a high-speed centrifuge after balancing, and the centrifugation conditions were: speed 6000rpm, time 20min. After the centrifugation, carefully remove the supernatant, pay attention not to lose the precipitate as much as possible, then add water to restore the volume, stir well and continue centrifuging, the centrifugation conditions are: 6000rpm, time 20min. After centrifugation, discard the supernatant, restore the volume of the bacteria to 250mL, stir evenly, pour the washed bacteria solution into a 500mL beaker, and prepare to break the wall.

[0065] (2) Cell wall breaking and PHA release of bacterial liqui...

Embodiment 3

[0077] Embodiment 3: superheated water, protease plus bleach method extract and purify PHA from the fermented liquid containing PHA thallus

[0078] method of extraction:

[0079] (1) Centrifugation and resuspension of bacteria for desalting

[0080] Halomonas containing 60% PHA (P3HB, P34HB can be) 1 250mL of the fermentation broth of the bacteria was placed symmetrically in a high-speed centrifuge after balancing, and the centrifugation conditions were: speed 6000rpm, time 20min. After the centrifugation, carefully remove the supernatant, pay attention not to lose the precipitate as much as possible, then add water to restore the volume, stir well and continue centrifuging, the centrifugation conditions are: 6000rpm, time 20min. After centrifugation, discard the supernatant, restore the volume of the bacteria to 250mL, stir evenly, pour the washed bacteria solution into a 500mL beaker, and prepare to break the wall.

[0081] (2) Cell wall breaking and PHA release of bacte...

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Abstract

The existing extraction and purification method of polyhydroxyalkanoate (PHA) comprises: adding hydrolase and the like into an organic solvent or a surfactant, and is complex and expensive in process.According to the invention, according to the method, a PHA-rich bacterial fermentation broth is treated with hot water or superheated water with a temperature of more than 100 DEG C so as to completewall breaking and purification of bacterial cells, and surfactant, protease and / or bleaching agent treatment and the like can also be combined to further improve the purity of PHA; and the method hasthe advantages of low cost, simple operation and high product recovery rate, and overcomes the problems of complex steps, high cost of addition of chemical substances, difficult treatment of downstream sewage and the like of the traditional PHA extraction.

Description

technical field [0001] The invention belongs to the downstream post-treatment field of bioengineering, in particular to the production and downstream extraction and purification process of polyhydroxyalkanoate (PHA). Background technique [0002] Polyhydroxyalkanoates (PHA) is a degradable polyester that can be accumulated by a variety of bacteria. According to its monomer composition, it can be divided into short-chain / or medium- and long-chain PHA, and there are also homopolymers and copolymers. . PHA is an environmentally friendly material with broad application prospects as an environmentally friendly substitute for traditional petroleum-based plastics. Among them, poly-3-hydroxybutyrate (poly-3-hydroxybutyrate, PHB) is currently the most clearly studied among the members of the PHA family, but its application is limited by its high crystallinity, low elasticity, and poor flexibility. . At present, the method of biological fermentation is often used in industry 7,8 ,...

Claims

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Application Information

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IPC IPC(8): C08G63/90C08G63/08
CPCC08G63/08C08G63/90
Inventor 陈国强张李湛叶健文吴赴清司徒卫
Owner TSINGHUA UNIV
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