Method for breeding epsilon-poly-L-lysine tolerant high-producing strain

A polylysine, high-yielding strain technology, applied in microorganism-based methods, biochemical equipment and methods, hybrid cell preparation, etc., can solve the problem of consuming a lot of time and energy

Inactive Publication Date: 2015-07-22
JIANGNAN UNIV
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  • Summary
  • Abstract
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Problems solved by technology

[0004] Mutants of strains can be obtained by classical chemical mutagenesis, but this method usually requires a lot of time and effort
Although genetic engi...

Method used

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Embodiment 1

[0017] 1) Construction of mutant strain library: at 1-5×10 8 Streptomyces spores / mL solution was added DES with a volume ratio concentration of 2.0%, treated at 30°C for 30 minutes, coated with a plate containing 0.48g / Lε-PL, cultured for 7-9 days, and selected spore formation speed, color The strains with darker, thicker spore layer and darker back color are numbered and screened first, and 10-20 strains with high yields are re-screened, and the strains with the highest yields are selected for the next round of DES mutagenesis. At the same time, the ε -PL concentration rose to 0.54g / L. By analogy, three rounds of chemical mutagenesis were carried out, and the concentration of ε-PL was increased to 0.60g / L. The 10 strains with higher yield than the parent were obtained for re-screening, and the 4 strains with the highest ε-PL yield were selected as the starting strains for protoplast fusion.

[0018] 2) Protoplast preparation, fusion and regeneration: the bacterial strain ob...

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Abstract

The invention belongs to the field of microbial breeding and relates to a method for screening an epsilon-poly-L-lysine tolerant high-producing strain through restructuring a genome. The method is characterized in that 16g/L epsilon-poly-L-lysine is added in a restructured strain regeneration medium by using a 4-5 cycle progressive protoplast fusion method, and finally, the yield-increased restructured strain capable of growing at high concentration is obtained.

Description

technical field [0001] The invention belongs to the field of microbial breeding, and relates to a method for genome shuffling and screening of ε-polylysine-tolerant high-yield strains. Background technique [0002] ε-polylysine (ε-poly-l-lysine, ε-PL) is a microbial secondary metabolite formed by linking α-carboxyl and ε-amino groups in lysine monomers. Compared with chemically synthesized α-poly-l-lysine (α-poly-l-lysine, α-PL), ε-PL has the advantages of wider antibacterial spectrum, good biocompatibility and non-toxic and harmless. As a food preservative, it has been approved for use in Japan, the United States, South Korea and China, and the demand is increasing day by day. In addition, as a gene carrier and cell culture material, it can inhibit the activity of pancreatic lipase in the small intestine, hinder the absorption of fat in the diet, and reduce obesity and other fields of medicine and biotechnology. It has potential application prospects. [0003] However, ac...

Claims

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Application Information

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IPC IPC(8): C12N15/03C12R1/465
Inventor 唐蕾周永鹏张建华张宏建毛忠贵
Owner JIANGNAN UNIV
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