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Succinic acid-producing strain and its screening method and uses

A technology for producing succinic acid and succinic acid, which is applied in the field of bioengineering and can solve problems such as difficult cultivation

Inactive Publication Date: 2006-12-27
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Typical gastrointestinal bacteria: E.coli, Pectinatus sp., Bacteroides sp.; rumen bacteria such as: Mannheimia succiniciproducens MBEL55E, Prevotella ruminicola (ATCC 19188), Succinimonas amylolytica (ATCC 19716), Ruminobacter amylophilus (DSM 1361), can accumulate D Diacid strains, but some of these strains are strictly anaerobic bacteria, which are relatively strict to equipment and culture conditions, and are not easy to cultivate. Some acid production levels are too low, and the concentration of substrates and products is too high. There are some deficiencies in industrialized production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Weigh 1.0 g of the strain-containing mixture obtained from the rumen of the cow, and add it to 50 ml of sterile water to prepare a rumen bacteria suspension. Take 1ml of rumen bacteria suspension into a 100ml serum bottle, the amount of enrichment medium in the serum bottle is 50ml, and the composition of the enrichment medium is: NaCl 1.0g / L, K 2 HPO 4 4.0g / L, MgCl 2 ·6H 2 O 0.05g / L, CaCl 2 ·H 2 O0.05g / L, (NH 4 ) 2 SO 4 0.2g / L, yeast extract 10g / L, peptone 5.0g / L, glucose 50.0g / L, monensin 0.2mg / L, disodium fumarate 20.0g / L, disodium succinate 60.0 g / L, magnesium carbonate 50g / L; after enrichment culture at 37°C for 18h, the culture sample was diluted 10 -6 , take a 0.1ml droplet spread on the flat plate containing the separation medium, and cultivate it in an anaerobic incubator. 12 20mg / L, lipoic acid 20mg / L, niacin 20mg / L, pantothenic acid 20mg / L, thiamine 20mg / L, vitamin B 6 20mg / L, agar 20g / L; mixed gas N in the anaerobic incubator 2 :CO 2 :H 2 =8:1...

Embodiment 2

[0049] Seed medium: yeast extract 5g / L, peptone 5g / L, NaCl 1g / L, NaHCO 3 10g / L, Na 2 HPO 4 9~15g / L,KH 2 PO 4 4~12g / L, add agar 20g / L when cultivating on a slant surface, and inject N 2 :CO 2 = 4:1 mixed gas for 2 minutes, sterilized at 121°C for 20 minutes, and cooled.

[0050] Fermentation medium: glucose 100g / L, MnCl 2 0.05g / L, FeCl 2 0.05g / L, CaCl 2 0.05g / L, K 2 HPO 4 2g / L, MgCO 3 100g / L, vitamins added folic acid 20mg / L, biotin 20mg / L, vitamin B 12 20mg / L, lipoic acid 20mg / L, niacin 20mg / L, pantothenic acid 20mg / L, thiamine 20mg / L, vitamin B 6 20mg / L, yeast extract and peptone respectively use the following dosages:

[0051] (1) 10g / L yeast extract, 5g / L peptone;

[0052] (2) 15g / L yeast extract, 5g / L peptone;

[0053] (3) 10g / L yeast extract, 15g / L peptone;

[0054] The volume of 100ml serum bottle is 50ml, and N 2 :CO 2 = 4:1 mixed gas for 2 minutes, sterilized at 121°C for 20 minutes, and cooled.

[0055] After streaking the strain with the pre...

Embodiment 3

[0059] Seed medium: yeast extract 5g / L, peptone 5g / L, NaCl 1g / L, NaHCO 3 10g / L, Na 2 HPO 4 9~15g / L,KH 2 PO 4 4~12g / L, (add agar 20g / L when cultivating on a slant), pass through N 2 :CO 2 = 4:1 mixed gas for 2 minutes, sterilized at 121°C for 20 minutes, and cooled.

[0060] Fermentation medium: glucose 100g / L, yeast extract 10-50g / L, peptone 10-50g / L, MnCl 2 0.05g / L, FeCl 2 0.05g / L, CaCl 2 0.05g / L, K 2 HPO 4 2.0g / L, MgCO 3 100g / L, vitamins added folic acid 5mg / L, biotin 5mg / L, vitamin B 12 5mg / L, lipoic acid 5mg / L, niacin 5mg / L, pantothenic acid 5mg / L, thiamine 5mg / L, vitamin B 6 5mg / L, pH 7.0.

[0061]When cultivating in a 5L fermenter, the liquid volume of the medium is 3L, with 100g / L MgCO 3 As a pH buffer, the fermenter gas mixture (N 2 :CO 2 =4:1) the ventilation rate is 0.6L / min, to maintain the anaerobic environment of the fermentation system. The strain with the preservation number CGMCC No.1716 was streaked on the seed slant medium, and then c...

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Abstract

The invention discloses a strain which can produce amber acid and its sieve method and its application. The said strain is named by classification as Actinobacillus succinogenes NJ113 and has been preserved in China microbial bacterial preservation management committee common microorganism center with CGMCC No.1716 as its preserved number. The invention prepares the ray bacillus which can produce amber acid with a high yield by enrichment culture of strain in animal cud in culture medium containing disodium fumarate and the following classification. The amber acid with higher concentration can be produced by performing fermentation by using the said strain, which is suitable for the commercial manufacture of amber acid.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a strain producing succinic acid. The invention also relates to a screening method for the strain and a method for fermenting and producing succinic acid using the strain. Background technique [0002] As an important chemical raw material, succinic acid (also known as succinic acid) is widely used in industries such as medicine, food and surfactant. However, the traditional chemical synthesis of succinic acid has limited the wide application of succinic acid as a basic chemical raw material due to reasons such as cost and environmental pollution. Because of this, the research and development of succinic acid production by microbial fermentation is being carried out in depth in the United States, Japan and other countries. [0003] The production of succinic acid by fermentation has many advantages. First of all, the new microbial fermentation production process and the ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/46
Inventor 姜岷韦萍陈可泉苏溧蔡婷王倩楠欧阳平凯
Owner NANJING UNIV OF TECH
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