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Strain for production of L-serine and method for production of L-serine by using same

A technology of serine and Corynebacterium glutamicum, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of difficult process amplification and industrialization, etc., and achieve increased income and stable yield traits , good effect of genetic stability

Active Publication Date: 2007-05-23
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using such recombinant bacteria to produce L-serine, it is difficult to scale up the process and realize industrialization

Method used

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  • Strain for production of L-serine and method for production of L-serine by using same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Screening of strains with L-serine synthesis ability.

[0028] Collect soil samples below 15cm below the surface around the sugar factory. 1g soil samples are enriched in a conical flask (30mL / 250mL) of enrichment medium containing 50μg / mL nystatin in a reciprocating shaker at 30°C 110rpm / min. The sets were cultured for 3 days. Take 0.1 mL of the enriched bacterial suspension and spread it on the screening medium, use a sterilized filter paper sheet to copy the coated plate to a copy plate, and cultivate for 3 days. Use 0.3% ninhydrin in acetone solution to infiltrate the plate, and dry it at 105°C for 10 minutes to develop the color. Those that produce purple color circles are amino acid producing bacteria.

[0029] Pick the corresponding amino acid-producing bacteria from the copy plate, and perform preliminary screening in the fermentation medium by shaking tubes, one tube for each strain. After the cultivation, the content of L-serine in the fermentation...

Embodiment 2

[0034] Embodiment 2 selects the optimum carbon source type and concentration of the fermentation medium.

[0035] Using the fermentation medium in Example 1 as the basal medium, the glucose therein was replaced with carbon sources such as 80 g / L sucrose, fructose, lactose, maltose, starch, etc. The results are shown in Table 2. When glucose, sucrose or fructose were used as carbon sources, the yield of L-serine was higher; when lactose, maltose, and soluble starch were used as the only carbon sources, almost no L-serine was produced. Among them, when sucrose was used as carbon source, the yield of L-serine was the highest, which was 5.1g / mL.

[0036] Carbon source (80g / L)

L-serine (g / L)

Residual sugar (g / L)

Conversion rates(%)

glucose

sucrose

fructose

lactose

Maltose

soluble starch

2.9

5.1

3.3

0

0

0

21.6

17.2

19.3

43.2

41.2

19.4

5.87

8.12

5.4

0

0

0

...

Embodiment 4

[0039] Example 4 VB 1 It has a great influence on bacterial growth and L-serine accumulation.

[0040] In the fermentation medium: sucrose 60g / L, corn steep liquor 10-30g / L, cottonseed powder 10-30g / L, (NH 4 ) 2 SO 4 20~50g / L,KH 2 PO 4 1.0~2.0g / L, MgSO 4 ·7H 2 O 0.5~1.0g / L, FeSO 4 ·7H 2 O 0.01~0.05g / L, MnSO 4 ·H 2 O 0.01~0.05g / L, biotin 0.08~0.15mg / L, CaCO 3 On the basis of 20g / L, add 50μg / L, 200μg / L, 500μg / L, 800μg / L of VB 1 , the ventilation rate of the fermenter is 2-3vvm, the stirring speed is 400-600 rpm, and the pH of the culture is within the range of 6-8. The results showed that 500μg / L was the optimal concentration, and the yield of L-serine could reach 6.0-6.4g / L. with and without VB 1 A comparative increase of about 20%.

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Abstract

A method for producing L-serine with a strain capable of producing L-serine belongs to the field of producing serine with biological method. The invention comprises the steps of: enriching, separating, detecting with discoloration halos of ninhydrin, preliminarily screening, and screening to obtain Corynebacterium glutamicum SYPS-062 (CGMCC No.1843) capable of producing L-serine, and the method for producing L-serine by directly fermenting saccharine material with the strain. The level of producing L-serine can reach to 6g / L at optimum conditions.

Description

technical field [0001] A bacterial strain producing L-serine and a method for producing L-serine by using the bacterial strain. The invention relates to a method for producing L-serine by using the bacterial strain to ferment and produce L-serine with sucrose and ammonium sulfate as main raw materials. It belongs to the technical field of amino acid preparation by biological method. Background technique [0002] L-serine is a non-essential amino acid and a sugar-producing amino acid. It has important biological significance and is a precursor for the synthesis of various amino acids, purine, thymine, and choline, and its derivative phosphatidylserine is one of the main components of phospholipids. It is widely used in the fields of food, medicine and cosmetics, as the main component of amino acid infusions, and as anticancer drugs, anti-AIDS drugs, etc. has attracted people's attention in the field of medicine; in the field of cosmetics, as a natural moisturizing Factors a...

Claims

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Application Information

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IPC IPC(8): C12P13/06C12N1/20C12R1/15
Inventor 许正宏张晓娟许泓瑜窦文芳
Owner JIANGNAN UNIV
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