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Method for improving yield of fermentation of epsilon-polylysine

A polylysine and yield technology, applied in the field of increasing the fermentation yield of ε-polylysine, can solve the problems of restricting further research and utilization, low capacity, etc., and achieve the effect of increasing positive mutation rate and improving fermentation level

Inactive Publication Date: 2011-08-17
ANHUI BBCA FERMENTATION TECH ENG RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Japan is the main country in the production of ε-polylysine by biological fermentation. Mutagenesis breeding is an effective method for breeding high-yield mutant strains. Mutagenesis breeding includes two steps of mutagenesis and screening. Mutations occur, and the strains screened from the mutant population are subjected to mutagenesis, and then screened again. Through the alternation of mutagenesis and screening, high-yield strains are selected, and the strains of Streptomyces albicans isolated by natural screening produce ε-polylysine Acid capacity is very low, thus limiting its further research and utilization

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] 1) Plate screening culture: Streptomyces albulus (CGMCC4.121) was inoculated on a plate screening medium under sterile conditions, and cultured at a constant temperature of 28°C for 7-8 days.

[0015] 2) Select 80 strains of the strains cultivated in step 1) and inoculate them into fermentation bottles under aseptic conditions, and culture them at a constant temperature for 8 days at 28° C. and 300 r / min.

[0016] The screening plate medium formula (g / L) used in step 1) is: glucose 10, yeast extract 4, wheat germ 1, agar powder 20.

[0017] Wherein step 2) the fermentation medium formula (g / L) that uses: glucose 50, (NH 4 ) 2 SO 4 10,K 2 HPO 4 1.6, KH 2 PO 4 0.8, MgSO 4 0.5, yeast paste 5, pH 6.8.

[0018] Fermentation culture process: under the condition of 28°C, 300r / min, constant temperature culture for 8 days, the average fermentation level reached 0.60g / L.

Embodiment 2

[0020] 1) Streptomyces albicans was mixed with normal saline to form a concentration of 10 6 cells / mL. Vibrated at 200r / min for 1 hour, then microwaved for 20s, treated with diethyl sulfate for 60min, and immediately spread on a screening plate for cultivation.

[0021] 2) Plate screening culture: Inoculate Streptomyces albulus (CGMCC 4.121) undergoing multiple mutagenesis on the screening plate medium under aseptic conditions, and culture at a constant temperature of 28°C for 7-8 days.

[0022] 3) Randomly select 80 strains of the strains cultivated in step 2) and inoculate them into fermentation bottles under aseptic conditions, and culture them at constant temperature for 8 days at 28° C. and 300 r / min.

[0023] Wherein step 2) the formula (g / L) of screening plate medium used: glucose 10, yeast extract 4, wheat germ 1, agar powder 20.

[0024] Wherein step 3) the fermentation medium formula (g / L) that uses: glucose 50, (NH 4 ) 2 SO 4 10,K 2 HPO 4 1.6, KH 2 PO 4 ...

Embodiment 3

[0027] 1) Streptomyces albicans was mixed with normal saline to form a concentration of 10 8 cells / mL. Vibrated at 200 rpm for 1 hour, then microwaved for 40 seconds, treated with diethyl acetate for 80 minutes, and immediately spread on a screening plate for cultivation.

[0028] 2) Plate screening culture: Streptomyces albulus (CGMCC 4.121) undergoing compound mutagenesis was inoculated on a plate screening medium under aseptic conditions, and cultured at a constant temperature of 28°C for 78 days.

[0029] 3) Randomly select 80 strains of the strains cultivated in step 2) and inoculate them into fermentation bottles under aseptic conditions, and culture them at constant temperature for 8 days at 28° C. and 300 r / min.

[0030] The formula (g / L) of the plate screening medium used in step 2): glucose 10, yeast extract 4, wheat germ 1, agar powder 20.

[0031] Wherein step 3) the fermentation medium formula (g / L) that uses: glucose 50, (NH 4 ) 2 SO 4 10,K 2 HPO 4 1.6, ...

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Abstract

The invention provides a method for improving yield of fermentation of epsilon-polylysine. In the method, a microwave and dithyl sulfate combined mutation induction method is used to obviously improve the forward mutation of Streptomyces albulus CGMCC4.121 to obtain a high-yield strain for producing epsilon-polylysine by fermentation. Results show that the shake flask fermentation level of the epsilon-polylysine is improved from 0.60g / L to 0.85g / L.

Description

technical field [0001] The invention relates to the field of mutation breeding and microbial fermentation, in particular to a method for increasing the fermentation yield of ε-polylysine. Background technique [0002] ε-polylysine (ε-polylysine, ε-PL) is a new, safe and efficient antiseptic preservative first discovered by Japan in the 1980s. It has inhibitory effect on Gram-positive bacteria and Gram-negative bacteria, yeast and other fungi and viruses under neutral and slightly acidic conditions, and has characteristics such as broad antibacterial spectrum. In developed countries such as Europe, America and Japan, ε-polylysine has been approved as a food preservative, mainly used in meat products, high-salt food, fast food, salad, cake and pastry food. Since ε-polylysine is a nutritional antibacterial agent with a wide range of antibacterial, it has become a high value-added biological antiseptic preservative. In addition, ε-polylysine is rich in cations and is widely us...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N15/01C12N13/00C12R1/47
Inventor 李荣杰张海涛
Owner ANHUI BBCA FERMENTATION TECH ENG RES
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