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Method for preparing polyhydroxybutyrate (PHB)

A technology of polyhydroxybutyrate and bacteria, applied in the biological field, can solve the problems of low economic benefit and the like

Inactive Publication Date: 2013-08-14
BIOCHEM ENG COLLEGE OF BEIJING UNION UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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

Method used

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  • Method for preparing polyhydroxybutyrate (PHB)
  • Method for preparing polyhydroxybutyrate (PHB)
  • Method for preparing polyhydroxybutyrate (PHB)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The formula of the culture medium used in this example is:

[0069] Slant medium: yeast extract 10g / L, peptone 10g / L, (NH 4 ) 2 SO 4 5g / L, agar 15g / L, pH7.

[0070] Seed medium: yeast extract 10g / L, peptone 10g / L, (NH 4 ) 2 SO 4 5g / L, pH7.

[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 cultivate at 30°C for 24h;

[0074] b. Inoculate a ring of well-grown slanted culture into a 500 mL conical flask containing 100 mL of seed medium, the shake flask rotating speed is 180 r / min, and the temperature is 30 °C for 24 hours;

[0075] c. According to 10% of the inoculum, it is inserted into the fermentation medium, the capacity of the fermenter is 2L, the volume of liquid filling is 1.5L, the stirring speed in the fermenter is 40...

Embodiment 2

[0089] The formula of the culture medium used in this example is:

[0090] Slant medium: yeast extract 10g / L, peptone 10g / L, (NH 4 ) 2 SO 4 5g / L, agar 15g / L, pH7.

[0091] Seed medium: yeast extract 10g / L, peptone 10g / L, (NH 4 ) 2 SO 4 5g / L, pH7.

[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 cultivate at 30°C for 24h;

[0095] b. Inoculate a ring of well-grown slanted culture into a 500 mL conical flask containing 100 mL of seed medium, the shake flask rotating speed is 180 r / min, and the temperature is 30 °C for 24 hours;

[0096] c. According to 10% of the inoculum, it is inserted into the fermentation medium, the capacity of the fermenter is 2L, the volume of liquid filling is 1.5L, the stirring speed in the fermenter is...

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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 present invention relates to the field of biotechnology, in particular, to a method for preparing PHB by using Cupriavidus sp. strain sh-1. Background technique [0002] Polyhydroxyalkanoates (PHA) is an encapsulated polyester that can be synthesized by many kinds of microorganisms, and acts as a carbon source and energy storage material in microbial cells. Under nutrient deprivation, microorganisms degrade PHA for use as a carbon source and energy. PHA is a biosynthetic and biodegradable polymer. PHA has similar material properties to traditional plastics synthesized from petroleum, such as polyethylene and polypropylene, but can be synthesized with renewable energy (such as carbohydrates, fatty acids, etc. produced by plant photosynthesis), and can Completely degraded into the ecological cycle of nature, so it is considered a "green plastic", which can replace non-degradable traditional plastics, and has attracted widespread attention from the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 赵有玺龚平秦京花田恒奇姜林
Owner BIOCHEM ENG COLLEGE OF BEIJING UNION UNIV
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