Method for producing copolymer PHBHHx by metabolism control of aeromonas hydrophila

A technology of Aeromonas hydrophila and metabolic regulation, applied in the field of microbiology and bioengineering, to achieve the effect of solving the problem of high production cost, low cost, and superior material properties

Inactive Publication Date: 2010-04-21
铁岭鹿宝堂药业集团有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention aims at the problem that wild-type Aeromonas hydrophila cannot use glucose to synthesize PHA, and through metabolic regulation, realizes that wild-type Aeromonas hydrophila uses glucose to synthesize PHBHHx

Method used

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  • Method for producing copolymer PHBHHx by metabolism control of aeromonas hydrophila

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Metabolic regulation of Aeromonas hydrophila to synthesize PHBHH with different 3HHx monomer composition

[0016] Strain: Aeromonas hydrophila CGMCC 0911 (purchased from CGMCC)

[0017] Medium: 1. Seed LB medium:

[0018] Yeast powder 5.0g / L,

[0019] Peptone 10.0g / L,

[0020] NaCl 10.0g / L.

[0021] 2. Microbial mineral salt medium (MM):

[0022] Component I (20×): Na 2 HPO 4 ·12H 2 O(180g / L) and KH 2 PO 4 (30g / L)

[0023] Component II: (NH 4 ) 2 SO 4 (1g / L) and MgSO 4 ·7H 2 O(0.41g / L)

[0024] Component III (100×): Fe(III)-NH 4 -Citrate (17%) (5g / L) and CaCl 2 ·2H 2 O(2g / L)

[0025] Component IV (1000×): ZnSO 4 ·7H 2 O(100mg / L), MnCl 2 ·4H 2 O(30mg / L), H 3 BO 3 (300mg / L), CoCl 2 ·6H 2 O(200mg / L), CuSO 4 ·5H 2 O(10mg / L), NiCl 2 ·6H 2 O(20mg / L) and NaMoO 4 ·2H 2 O(30mg / L).

[0026] Sterilization method: use component II as the substrate during sterilization, and sterilize component I, component III and component IV with mother liquor respectively, and mix before culturing. Ir...

Embodiment 2

[0039] Example 2 Comparative test

[0040] Strain: Aeromonas hydrophila CGMCC 0911 (purchased from CGMCC)

[0041] Medium: Same as Example 1.

[0042] Culture conditions: the same as in Example 1.

[0043] PHA detection and analysis method: same as Example 1.

[0044] Fermentation tank: Preparation of fermentation medium: Take the configuration of 100ml of microbial mineral salt medium as an example, add component I (5ml), component III (1ml) and component IV to component II (93.9ml) after sterilization. 0.1ml), mix well, and use it as a fermentation medium. Inoculate Aeromonas hydrophila into the fermentation medium, the inoculation amount is 5-10% (v / v), and use dilute H 2 SO 4 Adjust pH 7.0 with ammonia water (or 1N sodium hydroxide), continue fermentation at 30°C for 48-72 hours, dissolved oxygen D.O. is automatically controlled by stirring, and the stirring speed control range is 200-500r / min. The carbon source is only lauric acid or glucose.

[0045] After the incubation, the co...

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Abstract

The invention relates to a method for producing a copolymer PHBHHx by metabolism control of aeromonas hydrophila, which comprises the steps of: inoculating the aeromonas hydrophila into a fermentation medium, adding a carbon source and acyl CoA, regulating the pH value to 7.0-7.2, keeping the temperature at 30 DEG C, controlling dissolved oxygen through stirring, controlling the stirring speed at 200-500r/min to enable the dissolved oxygen to be kept 10-20 percent, continuously fermenting for 48-72h, and synthesizing the copolymer PHBHHx containing different proportions of 3HHx according to demands. The carbon source comprises 5-20 percent of glucose and 0.5-2 percent of lauric acid in volume of the fermentation medium. By the proportion regulation of different carbon sources and the metabolism control of the aeromonas hydrophila, the PHBHH x containing 3HHx in different proportions can be synthesized according to the demands of different fields.

Description

Technical field [0001] The invention belongs to the field of microorganisms and bioengineering, and specifically relates to a method for producing a copolymer PHBHHx containing different proportions of 3HHx by metabolic regulation of Aeromonas hydrophila. Background technique [0002] Biomedical materials, also known as Biomaterials, are special functional materials that can diagnose, treat, replace, repair, induce, regenerate or enhance the functions of cells, tissues and organs in the body . Today, biomedical materials have evolved from the first and second generation biomedical materials in the 20th century to the third-generation biomedical materials based on the cellular and molecular levels. The third-generation biomedical materials can be degraded in the body and eventually absorbed by the body. At the same time, the materials themselves are biologically active. They can participate in the physiological activities of the body, activate genes at the molecular level, stimul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/62C12R1/01
Inventor 胡风庆回晶侯萧
Owner 铁岭鹿宝堂药业集团有限公司
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