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Streptomyces sp. for producing epsilon-polylysine and application of streptomyces sp.

A technology of polylysine and Streptomyces parvus, applied in the field of bioengineering, can solve the problems of unsuitable breeding work, general screening effect, expensive price, etc., and achieve the effect of excellent passage stability

Active Publication Date: 2020-01-07
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these "sieves" have mediocre screening effects and are very expensive, so they are not suitable for large-scale application in breeding work.

Method used

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  • Streptomyces sp. for producing epsilon-polylysine and application of streptomyces sp.
  • Streptomyces sp. for producing epsilon-polylysine and application of streptomyces sp.
  • Streptomyces sp. for producing epsilon-polylysine and application of streptomyces sp.

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: the breeding of Streptomyces parvus M-Z18

[0049] Bettner solid medium: 2% glucose, 0.2% yeast extract, 0.4% peptone, 2% agar, all in mass-volume fraction, pH 7.5, sterilized at 115°C for 20min.

[0050] Solid medium with succinic acid as carbon source: 2% succinic acid, 0.2% yeast extract, 0.4% peptone, 2% agar, all in mass-volume fraction, pH 7.5, sterilized at 115°C for 20 minutes.

[0051] Specific steps are as follows:

[0052] (1) Evenly spread 10-100 μL of spore suspension prepared by the wild fungus Streptomyces albicans Z-18 screened in the soil on a metal slide cooled by flame burning, and place in a normal temperature and pressure plasma (ARTP) mutagenesis system Operation (instrument power 100W, gas flow 10SLM) for 50-400s;

[0053] (2) Inoculate the spores of the wild fungus Z-18 treated in step (1) on the Bethner solid slant medium, and after cultivating at a constant temperature of 30°C for 5-8 days, suspend the spores in 5mL of phosphate...

Embodiment 2

[0067] Embodiment 2: Morphological characteristics of starting bacterium Z-18 and mutagenic Streptomyces albicans M-Z18

[0068] Through the identification of microbial morphological characteristics and physical and chemical characteristics, the characteristics of the starting strain Z-18 and the mutagenic strain M-Z18 were compared, as follows:

[0069] 1. Morphological characteristics

[0070] The aerial hyphae of the strain Z-18 grew well on the medium containing yeast extract or peptone, the spores were curvy, the spores were light gray chains, and were oval under the microscope; the colonies and their backs were yellow.

[0071] The aerial hyphae of the strain M-Z18 grow well on the peptone yeast extract medium, the spore filaments are spiral or wavy, and the spores are dark gray chains, which are oval under the microscope; the colonies and their backs are yellow or yellow brown.

[0072] 2. Cultivate characteristics

[0073] Culture at a constant temperature of 30°C f...

Embodiment 3

[0087] M3G seed medium: glucose 5%, (NH 4 ) 2 SO 4 1%, Na 2 HPO 4 0.14%, KH 2 PO 4 0.1%, MgSO 7 ·H 2 O 0.025%, ZnSO 7 ·H 2 O 0.005%, FeSO 7 ·H 2 O 0.001%, yeast powder 0.5%, pH 6.8, sterilized at 115°C for 20min.

[0088] RSM liquid fermentation medium: glycerol 6%, (NH 4 ) 2 SO 4 0.5%, beef extract 1%, KH 2 PO 4 0.4%, MgSO 4 0.08%, FeSO 4 0.005%, all are mass-volume fractions, sterilized at 115°C for 20 minutes, and fermented at natural pH.

[0089] The purified M-Z18 spores were inoculated into the seed medium filled with M3G, and cultured in shake flasks at 30° C.×200 rpm for 24 hours to prepare seed liquid. The seed solution was inoculated in the RSM fermentation medium with an inoculation amount of 8%, and the shake flask was fermented for 72 hours. The content of ε-PL in the fermentation supernatant of the mutant strain detected by methyl orange detection method was 1.8g / L, and the production efficiency of the strain was 0.45g / L / d. The cell mo...

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Abstract

The invention discloses streptomyces sp. for producing epsilon-polylysine and an application of the streptomyces sp., and belongs to the field of biotechnology. A streptomyces albulus (streptomyces albulus) M-Z18 bacterial strain is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO: M 2019589. The streptomyces albulus M-Z18 bacterialstrain can quickly grow on a culture medium using succinic acid as a carbon source, and has the capacity for high efficiency synthesis of epsilon-polylysine having sulfaguanidine resistance. The bacterial strain is subjected to fed-batch fermentation under a pH impact strategy, the accumulation concentration of the epsilon-polylysine can achieve 54.7g / L, the production efficiency can achieve 6.83g / L / d, and the streptomyces sp. is an epsilon-polylysine industrial production bacterial strain having great potential.

Description

technical field [0001] The invention relates to a strain of streptomyces producing ε-polylysine and its application, belonging to the technical field of bioengineering. [0002] technical background [0003] As a new type of natural biological food preservative, ε-polylysine (abbreviated as ε-PL) has a broad antibacterial spectrum, is easily soluble in water, has strong thermal stability, high safety, and is suitable for a wide range of pH values. In the 1980s, Japan first approved ε-PL as a food preservative. Subsequently, the United States, South Korea, Europe and China successively approved ε-PL as a new food additive. On the other hand, ε-PL, as a kind of cationic biopolymer, is widely used in the fields of biodegradable materials, drug carriers, biochip coatings, emulsifiers, highly absorbable hydrogels, and anticancer enhancers. and apply. Therefore, ε-PL is a new type of industrial biotechnology product with a wide range of applications and high added value, and has...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/465
CPCC12N1/20C12P13/02C12N1/205C12R2001/465
Inventor 陈旭升毛忠贵王靓张建华张宏建
Owner JIANGNAN UNIV
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