Method for extracting intracellular poly hydroxy fatty acid in microbe

A technology for polyhydroxyalkanoates and microorganisms, which is applied in the field of extracting intracellular polyhydroxyalkanoates of microorganisms, can solve the problems of difficult extraction, more impurities, difficult separation and drying, etc., and achieves broad application prospects and shortens production. time, good extraction effect

Inactive Publication Date: 2006-10-11
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above-mentioned extraction methods all have relatively large defects, such as low recovery rate, low product purity, and reduced product molecular weight after extraction. In addition, many organic solvents used are more toxic and have lower boiling points. The extraction process requires high pressure, which brings pressure to production. to safety hazard
For PHA with medium and long chains, the existing extraction methods usually result in the PHA precipitate being a very viscous substance, which is difficult to separate and dry, which brings certain difficulties to the extraction, and the purity is low, with more impurities, which limits the Its application in the field of medicine

Method used

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  • Method for extracting intracellular poly hydroxy fatty acid in microbe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1, extracting Aeromonas hydrophila 4AK4 intracellular polyhydroxyalkanoate

[0023] Now use n-butyl acetate from Aeromonas hydrophila 4AK4 (Chen GQ, G.Zhang, S.J.Park, S.Y.Lee, Industrial scale Production of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate).Appl Microbiol Biotechnol, 57(2001) 57 : 50-55) in dry thalline, extract short-chain and medium-long-chain copolymerization PHA (PHBHHx), wherein the structural formula of breaking the wall extraction solvent n-butyl acetate is as follows:

[0024] Formula 2

[0025] The specific extraction process includes the following steps:

[0026] 1) Weigh 20 g of fermented Aeromonas hydrophila 4AK4 dried thallus powder;

[0027] 2) Add 100mL of n-butyl acetate, mix well, stir at 150 rpm, and reflux heat treatment at 110°C for 1 hour;

[0028] 3) Centrifuge at 5000g for 10 minutes to remove the bacterial residue and keep the supernatant;

[0029] 4) Add 500mL 95% (v / v) alcohol to the supernatant, mix well, and let ...

Embodiment 2

[0033] Embodiment 2, extracting Aeromonas hydrophila 4AK4 intracellular polyhydroxyalkanoate

[0034] A mixed solvent of n-butyl acetate and isobutyl acetate is now used to extract short-chain, medium- and long-chain copolymerized PHA (PHBHHx) from the dry cells of Aeromonas hydrophila 4AK4, wherein the structural formula of isobutyl acetate in the wall-breaking extraction solvent is as follows:

[0035] Formula 3

[0036] The specific extraction process includes the following steps:

[0037] 1) Weigh 20 g of fermented Aeromonas hydrophila 4AK4 dried thallus powder;

[0038] 2) Add 60 mL of mixed solvent of n-butyl acetate and isobutyl acetate (volume ratio 1:1), mix well, stir at 200 rpm, and reflux heat treatment at 110°C for 1 hour;

[0039] 3) Centrifuge at 5000g for 10 minutes to remove the bacterial residue and keep the supernatant;

[0040] 4) Add 900mL 95% (v / v) alcohol to the supernatant, mix well, and let it stand for 30 minutes, white flocculent or powdery prec...

Embodiment 3

[0044] Embodiment 3, extracting Aeromonas hydrophila 4AK4 intracellular polyhydroxyalkanoate

[0045]Now use n-butyl acetate to extract short-chain and medium- and long-chain copolymerized PHA (PHBHHx) from the dried thallus of Aeromonas hydrophila 4AK4. The specific extraction process includes the following steps:

[0046] 1) Weigh 20 g of fermented Aeromonas hydrophila 4AK4 dried thallus powder;

[0047] 2) Add 300mL n-butyl acetate, mix well, stir at 130 rpm, and reflux heat treatment at 120°C for 1 hour;

[0048] 3) Centrifuge at 5000g for 10 minutes to remove the bacterial residue and keep the supernatant;

[0049] 4) Add 900mL 95% (v / v) alcohol to the supernatant, mix well, and let it stand for 30 minutes, white flocculent or powdery precipitates precipitate out;

[0050] 5) Centrifuge at 5000g for 2 minutes to collect the precipitate, soak and wash it with 60mL of absolute ethanol for 10 minutes, and centrifuge at 5000g for 5 minutes to collect the precipitate;

[00...

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Abstract

This invention is extraction method of cellular hydroxy fatty acid esters from microorganism. The procedure is as followings: 1) ferment to get enough microorganisms, collect it. 2) Dry to powder, add esterlike organic solvents. React at 80-120deg C for 1-4hours. Extract hydroxy fatty acid esters. 3) Restore the organic phase. 4) Precipitate hydroxy fatty acid esters from the organic phase with esterlike organic solvents. 5) Separate, wash and precipitate. 6) dry to obtain hydroxy fatty acid esters. This invention could be used in high quality PHA industrialization. It's time-reduced, energy-saving, environment-friendly and highly-efficient. It has a large application scale.

Description

technical field [0001] The invention relates to a method for extracting microbial intracellular polyhydroxyalkanoate. Background technique [0002] Polyhydroxyalkanoates (Poly-β-Hydroxyalkanoates, referred to as PHA) is a kind of intracellular polyester synthesized by microorganisms, which exists in the form of granular inclusions in microbial cells. The physical properties of PHA are similar to plastics from petrochemical sources, and it has biodegradable properties. It is considered as a potential "biodegradable plastic"; at present, research in the field of tissue engineering shows that PHA materials have good biocompatibility It is a very potential tissue engineering material. PHA exists in microorganisms in the form of inclusions. According to the PHA monomer (see formula 1 for the general structure, R1 and R2 can be different substituent groups. Typical structures of R1 and R2 groups are: CH 3 , (CH 2 )nCH 3 (n=1-8)) The composition is different, divided into short...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08G63/06C12P7/64
Inventor 陈国强欧阳少平
Owner TSINGHUA UNIV
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