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Actinobacillus and method for producing succinic acid

A technology of actinomycetes and production methods, applied in the field of bioengineering, to achieve the effect of improving biomass utilization and increasing yield

Inactive Publication Date: 2010-09-29
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most bacterial strains cannot meet the economic requirements of industrial production in production, and the lack of excellent succinic acid fermentation strains has always been a serious bottleneck restricting the development of succinic acid fermentation industry. microbial strains

Method used

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  • Actinobacillus and method for producing succinic acid
  • Actinobacillus and method for producing succinic acid
  • Actinobacillus and method for producing succinic acid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1 Screening of succinic acid-producing bacterial strains of the present invention

[0045] This embodiment is a screening method for succinic acid-producing strains of the present invention, and the specific operations are as follows:

[0046] (1) Sampling: Gastric juice was sampled from the rumens of two adult bulls with permanent fistulas in Beijing Institute of Animal Husbandry and Veterinary Medicine.

[0047] (2) Enrichment: Take rumen gastric fluid and inoculate it into 100ml of enrichment medium supplemented with 100mg / L amphotericin B and 80mg / L salinomycin, at 37°C in CO 2 Anaerobic culture in an environment for 24-48 hours, enriched twice according to the inoculum size of 5%; wherein, the enrichment medium used consists of: glucose 10g / L, yeast extract 10g / L, Na 2 HPO 4 12H 2 O 3.3g / L, NaH 2 PO 4 2H 2 O4.4g / L, CaCl 2 2H 2 O 1g / L, MgCl 2 ·6H 2 O 1g / L, MnCl 2 0.1g / L, MgCO 3 10g / L, medium pH3-8, sterilized at 115°C for 30 minutes, and cooled...

Embodiment 2

[0053] Example 2 Physiological and biochemical identification of CGMCC2650 strain

[0054] The 1600 times microscope photo of Actinobacillus succinogenes CGMCC 2650 is as follows figure 1 shown, from figure 1 It can be seen from the figure that the bacteria are short rod-shaped, the cells are cohesive, and there are no spores.

[0055] In this embodiment, according to the physiological and biochemical identification method in "Bergey's Bacteria Identification Handbook" (Ninth Edition) (Baltimore: TheWilliams and Wilkins Co. Baltimore Md publication, May 2004), the physiological and biochemical identification of the CGMCC2650 bacterial strain is carried out, The results are shown in Table 1 and Table 2.

[0056] Table 1 Physiological and biochemical identification results of CGMCC2650

[0057]

[0058]

[0059] Table 2 Carbon source distribution of CGMCC2650

[0060]

[0061] "+" is a positive response, "-" is a negative response

Embodiment 3

[0062] Example 3 Alignment of 16S rDNA sequences of CGMCC2650 strain

[0063] In this example, the total DNA of BE-1 bacteria was extracted, primers were designed to amplify its 16S rDNA gene, and then sequenced, and the homology data were compared in the NCBI database.

[0064] 16S rDNA gene amplification and PCR product analysis template preparation: absorb an appropriate amount of bacteria and centrifuge, take out the bacteria pellet, and use a genome extraction kit (for genomic DNA extraction methods, please refer to "Bacterial Molecular Genetics Classification and Identification Method" (Shanghai Science and Technology Co., Ltd.) Publishing House, 1990) to extract the total DNA of the bacteria, the fragment size is about 20k bp, and the electrophoresis results are as follows: figure 2 shown.

[0065] 16S rDNA gene primers were designed using universal primer 1, F27 (SEQ.ID.NO.1, 5'-AGAGTTTGATCATGGCTCAG-3') and primer 2, R1492 (SEQ.ID.NO.2, 5'-TACGGTTACCTTGTTACGACTT-3'...

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Abstract

The invention provides actinobacillus and a method for producing succinic acid. The Actinobacillus succinogenes CGMCC2560 for producing the succinic acid by fermenting saccharine materials can produce the succinic acid through fermentation, or joint production is performed through succinic acid fermentation and cellulosic ethanol fermentation. Under the anaerobic condition of introducing CO2 and / or N2 and in the environment that pH is maintained 5.5-7.5, 10-110g / L of succinic acid can be produced from 10-100g / L of saccharine materials by anaerobic fermentation. In the joint production method, CO2 generated in the cellulosic ethanol fermentation process is used for the fermentation production of the succinic acid so as to improve the yield of the succinic acid and the cellulosic ethanol and realize zero emission of the CO2. In the joint production, the yield of the succinic acid can reach 92g / L, and the yield of the cellulosic ethanol reaches 108g / L.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to an actinobacillus for producing succinic acid, a method for producing succinic acid by using the actinobacillus, and a combined production method and device for succinic acid and ethanol. Background technique [0002] Succinic acid (Succinic Acid, Butanedioic Acid), also known as succinic acid, is recognized by the US FDA as "generally recognized as safe" (GRAS), so it can be used for a variety of purposes. A 2004 report by the U.S. Department of Energy pointed out that succinic acid is a platform chemical substance for the synthesis of bulk daily necessities and important chemicals, and has huge economic value. With the shortage of energy, the prospect of producing succinic acid by biological fermentation is very broad. [0003] Succinic acid is a metabolic intermediate of many strict anaerobic bacteria and facultative anaerobic bacteria. Microorganisms that produce succin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/46C12P7/10C12M1/00C12R1/04
CPCC12M21/12Y02E50/16C12M43/04C12M23/58Y02E50/10
Inventor 邢建民李强李望良苏志国
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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