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
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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;
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