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A technology of polyhydroxybutyrate and seeds is applied in the field of preparing PHB by copper bacteria strain sh-1, which can solve the problems of low economic benefit and the like
Inactive Publication Date: 2011-10-26
BIOCHEM ENG COLLEGE OF BEIJING UNION UNIV
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Problems solved by technology
Existing production strains include Eutropha rosenbergii, Alcaligenes giantis, Azotobacter victoria, etc., which can be used in the industrial production of PHB, but the economic benefits of production are relatively low
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Embodiment 1
[0068] The formula of culture medium used in the present embodiment is:
[0071] Fermentation medium: Na 2 HPO 4 ·7H 2 O 2g / L, KH 2 PO 4 2g / L, MgSO 4 ·7H 2 O 0.2g / L, ferric ammonium citrate 50mg / L, CaCl 2 2H 2 O 10mg / L, (NH 4 ) 2 SO 4 2g / L, glucose 60g / L, pH7.
[0072] 1. Production of PHB:
[0073] a. Inoculate the strains on the slant medium and culture at 30°C for 24h;
[0074] b. Inoculate a ring of well-grown slant culture into a 500mL Erlenmeyer flask containing 100mL seed medium, shake the flask at a rotational speed of 180r / min, and culture at 30°C for 24h;
[0075] c. Insert 10% of the inoculum into the fermentation medium, the capacity of the fermenter is 2L, the liquid volume is 1.5L, the stirring speed in the fermenter is 400r / min, the ventilation rate is 100L / min...
Embodiment 2
[0089] The formula of culture medium used in the present embodiment is:
[0092] Fermentation medium: Na 2 HPO 4 ·7H 2 O 6g / L, KH 2 PO 4 4g / L, MgSO 4 ·7H 2 O 0.5g / L, ferric ammonium citrate 75mg / L, CaCl 2 2H 2 O 25mg / L, (NH 4 ) 2 SO 4 7g / L, glucose 100g / L, pH7.
[0093] 1. Production of PHB:
[0094] a. Inoculate the strains on the slant medium and culture at 30°C for 24h;
[0095] b. Inoculate a ring of well-grown slant culture into a 500mL Erlenmeyer flask containing 100mL seed medium, shake the flask at a rotational speed of 180r / min, and culture at 30°C for 24h;
[0096] c. Insert 10% of the inoculum into the fermentation medium, the capacity of the fermenter is 2L, the liquid volume is 1.5L, the stirring speed in the fermenter is 400r / min, the ventilation rate is 180L / ...
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Abstract
The invention discloses a method for preparing polyhydroxybutyrate (PHB), which is implemented by taking cupriavidus strains sh-1 (preservation No.: CGMCC No. 4815) as strains for preparing PHB. The method comprises the following steps: inoculating the strains in a slant culture medium, then culturing the strains for 24 hours at a temperature of 30 DEG C; taking slant cultures, then inoculating the slant cultures in a conical flask filled with a seed culture medium, and under the condition of shaking the conical flask at a rotating speed of 180 r/min, culturing the slant cultures for 24-36 hours at a temperature of 30 DEG C; inoculating the obtained seed liquid in a fermentation medium according to an inoculation quantity of 10%, and stirring the obtained product at a rotating speed of 300-500 r/min, then after fermenting the seed liquid 72 hours at a ventilatory capacity of 50-200 L/min and a fermentation temperature of 28-35 DEG C, stopping the operation of fermentation; and in the process of fermentation, when the dissolved oxygen content of the fermentation medium is increased to 80% of saturated dissolved oxygen content, adding a glucose solution (concentration: 10-300 g/L) into the fermentation medium, then when the dissolved oxygen content of the obtained solution drops to 50%, stopping the operation of adding. By using the method disclosed by the invention, the high-density fermentation and high yield of polyhydroxybutyrate can be realized.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a method for preparing PHB with Cupriavidus sp. strain sh-1. Background technique [0002] Polyhydroxyalkanoates (PHA) is an in-package polyester that can be synthesized by many kinds of microorganisms, and is used as a carbon source and energy storage material in microbial cells. Under nutrient starvation, microorganisms degrade PHA for use as a carbon source and energy. PHA is a high molecular polymer that can be biosynthesized and biodegraded. PHA has similar material properties to traditional petroleum-based plastics such as polyethylene and polypropylene, but can be synthesized with renewable energy sources (such as carbohydrates and fatty acids produced by plant photosynthesis), and can It is completely degraded and enters the ecological cycle of nature, so it is considered a kind of "green plastic", which can replace non-degradable traditional plastics, and has attracted ext...
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