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Actinobacillus succinate producing strain and method for its screening and fermentative production of succinic acid

An actinomycetes, succinic acid-producing technology, applied in the field of bioengineering, can solve the problems of unreachable yield, difficult to obtain strains, low yield and so on

Inactive Publication Date: 2017-06-20
GUANGXI ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] To sum up, at present, due to foreign technical blockade, it is difficult to obtain foreign strains. Even if foreign strains are purchased, the specific technical conditions are difficult to know, and the yield is not high, which cannot reach the yield reported in the literature; There is still a certain gap between the production of domestic succinic acid strains and foreign countries. At present, the highest domestic production is about 50g / L succinic acid

Method used

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  • Actinobacillus succinate producing strain and method for its screening and fermentative production of succinic acid
  • Actinobacillus succinate producing strain and method for its screening and fermentative production of succinic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Screening and identification of strains of Actinobacillus succinate:

[0059] 1. Preparation of enrichment medium: a medium containing fumarate as the sole carbon source and adding nitrogen sources, inorganic salts and monensin. The detailed formula is as follows (g / L): sodium fumarate 10-30, yeast powder 5-20, peptone 5-20, K 2 HPO 4 1-5, CaCl 2 0.1-1, NaCl 0.5-2, (NH 4 ) 2 SO 4 0.5-2, MgCl 2 0.1-1, monensin 0.1-0.2, mix the aforementioned raw materials to obtain an enriched medium. It is verified by experiments that the enrichment medium prepared by any combination of concentrations in the aforementioned enrichment medium raw material formula can screen the GXAS-137 strain of the present invention.

[0060] 2. Preparation of plate screening medium: Add sodium succinate and agar powder on the basis of enriched medium. The detailed formula is as follows (g / L): sodium fumarate 10-30, yeast powder 5-20, peptone 5-20, K 2 HPO 4 1-5, CaCl 2 0.1-1, NaCl0.5-2, (NH 4 ) 2 SO 4 0.5-...

Embodiment 2

[0110] According to the method of fermentative production of succinic acid by Actinobacillus succinate-producing strain in Example 1, the plate of the strain with the deposit number CCTCCNO:M2011399 was cultured in an anaerobic box for 48 hours, and then a loop was connected to 30mL seed culture medium with an inoculation loop. Cultivate for 20 hours at 37°C, and access fermentation medium (250 mL Erlenmeyer flask with a liquid volume of 150 mL) of different carbon sources (65 g / L based on glucose) according to the 5% inoculum volume, and statically culture for 48 hours. The content of diacid and main by-products, the results are shown in Table 2.

[0111] Table 2 Effects of different carbon sources on the production of succinic acid by CCTCC NO: M2011399

[0112] Carbon source Succinic acid content (g / L) Acetic acid content (g / L) Lactic acid content (g / L) glucose47.9810.681.63 sucrose50.318.360.91 fructose40.349.271.39 maltose49.6710.831.50 Arabic candy54.767.241.37 Xyl...

Embodiment 3

[0115] According to the method for fermentative production of succinic acid by the strain of Actinobacillus succinate in Example 1, the plate of the strain with the deposit number CCTCCNO: M2011399 was cultured in an anaerobic box for 48 hours, and then a loop was connected to 30 mL of seed medium with an inoculum loop, 37 Cultivate for 20h at ℃, insert fermentation medium (250mL Erlenmeyer flask with a liquid volume of 150mL) of different raw materials (75g / L based on glucose) according to the 5% inoculum, static culture for 48h, and determine succinic acid by HPLC And the main by-product content, the results are shown in Table 3.

[0116] Table 3 Effects of different raw materials on the production of succinic acid by CCTCC NO: M2011399

[0117] Carbon source Succinic acid content (g / L) Acetic acid content (g / L) Lactic acid content (g / L) Tapioca flour70.1710.771.46 corn flour53.8112.342.01 Cane molasses49.377.261.17 Sugar cane juice58.678.911.64

[0118] It can be seen fr...

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Abstract

The invention discloses an actinobacillus succinogenes GXAS-137, and a method for producing succinic acid by screening and fermentation of the strain; the strain is obtained by enrichment and screening from a cattle rumen; the method comprises the following steps: inoculating the actinobacillus succinogenes strains in a seed medium, culturing for 24-36 hours in an anaerobic incubator or a common incubator at 30-38 DEG C in a static manner, inoculating into a fermentation medium with an inoculation amount of 1-10%, adjusting the pH to 6.0-7.0, and performing fermentation at 30-38 DEG C for 24-120 hours to obtain succinic acid. Fermentation research is performed with different raw materials and in different media; the succinic acid yield is up to 70 g / L; and the method has potential for industrial production.

Description

Technical field [0001] The invention relates to the technical field of bioengineering, in particular to the screening and application of a strain of Actinobacillus succinate-producing. Background technique [0002] Succinic acid, also known as succinic acid (succsinic acid), is a common natural organic acid that is widely found in animals, plants and microorganisms, and is one of the intermediate products of biological TCA cycle. It is an important organic synthesis intermediate, mainly used in food, medicine, biodegradable plastics, surfactants, detergents, and green solvents. In addition, succinic acid can also be used as animal feed additives and plant growth stimulators. At present, the production of succinic acid mainly uses the method of chemical synthesis. The production of succinic acid from butane through maleic anhydride is mainly by paraffin oxidation, light oil oxidation, butane oxidation, and succinonitrile hydrolysis. , Catalytic hydrogenation and other methods. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/46C12R1/01
CPCY02P20/141
Inventor 申乃坤黄日波王青艳秦艳黎贞崇朱婧王成华廖思明
Owner GUANGXI ACAD OF SCI
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