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Streptomyces albidoflavus and application thereof in production of epsilon-polylysine

A technology of Streptomyces albicans and polylysine, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of general screening effect, high price, unsuitable for large-scale application in breeding work, etc., to avoid The effects of bacterial contamination, strong product tolerance, and excellent passage stability

Active Publication Date: 2021-02-12
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A common method for breeding ε-PL high-yielding strains is to screen mutant strains with resistance to L-lysine structural analogues and product ε-PL resistance. However, these "sieves" have general screening effects and are very expensive. Not suitable for large-scale application in breeding work

Method used

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  • Streptomyces albidoflavus and application thereof in production of epsilon-polylysine
  • Streptomyces albidoflavus and application thereof in production of epsilon-polylysine
  • Streptomyces albidoflavus and application thereof in production of epsilon-polylysine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Embodiment 1: Breeding of Streptomyces parvus GS114

[0073] Bethner solid medium (g / 100mL): glucose 2, yeast extract 0.2, peptone 0.4, agar 2, pH 7.5, sterilized at 115°C for 20 minutes.

[0074] Antibiotic resistance medium (g / 100mL): glucose 2, yeast extract 0.2, peptone 0.4, agar 2, pH 7.5, sterilized at 115°C for 20 minutes. When the temperature of the culture medium dropped to 60°C, the stock solutions of streptomycin, gentamicin and rifamycin prepared in advance and sterilized by filtration through a 0.22 μm membrane were added thereto.

[0075] Specific steps are as follows:

[0076] (1) Construction of mutation library: 2% (v / v) DES was used to test the spores of Streptomyces albicans M-Z18 (the deposit number is CCTCC NO: M 2019589, and the strain is recorded in the patent application with application number 201911020454.4) The suspension was treated for 30min (28°C water bath), and NaS was added 2 o 3 Terminate the reaction, dilute and smear on a Bethner-...

Embodiment 2

[0078]Example 2: Morphological analysis of the starting bacterium M-Z18 and the high-yielding bacterium GS114

[0079] The morphology of the strains was observed with naked eyes and microscopic analysis.

[0080] Depend on figure 1 It can be seen that there are obvious differences in morphology between the two strains. On the peptone yeast extract medium, the colony of the starting strain M-Z18 was larger, the aerial hyphae grew well, and the colony and its back were yellow or tan. In YP liquid medium, the mycelium of M-Z18 intertwined to form larger solid balls.

[0081] The colonies of the strain GS114 are small. On the peptone yeast extract medium, the aerial hyphae grow well, and the spores form gray-green chains, which are oval under the microscope; the colonies and their backs are yellow-green or yellow-brown. In YP liquid medium, the hyphae of GS114 were relatively loose and the balls were small.

Embodiment 3

[0082] Example 3: Resistance of Streptomyces parvus GS114 to a single antibiotic

[0083] Bettner solid medium: glucose 20g / L, yeast extract 2g / L, peptone 4g / L, agar 20g / L, pH7.5, sterilized at 115°C for 20min.

[0084] Preparation of single spore suspension: Streptomyces albicans GS114 was streak-inoculated on Bethner's medium, cultured at 30°C for 8 days, and after the spores matured, the spores were placed in an Erlenmeyer flask containing glass beads and sterile water. Shake at 200r / min on a shaker at 30°C for 20min, filter through 8 layers of sterile gauze to obtain a single spore suspension, and count with a hemocytometer under an optical microscope to determine the concentration of the prepared single spore suspension, using an order of magnitude of 10 8 ~10 9 cells / mL was used as the spore suspension.

[0085] Streptomycin and gentamicin can bind to the ribosomal S12 protein and L6 protein, respectively, and the strain will die due to the inhibition of protein transl...

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Abstract

The invention discloses streptomyces albidoflavus and application thereof in production of epsilon polylysine, and belongs to the field of microbial fermentation engineering. The Streptomyces albus GS114 provided by the invention can grow on an agar culture medium containing more than 10 [mu] g / ml of one or more of streptomycin, gentamycin and rifamycin, shows very strong antibiotic resistance, can normally grow in a liquid culture medium with a pH value of 2.5-3.0 and can synthesize a large amount of epsilon PL, and after fermentation for 200 h, the Streptomyces albus GS114 can be used as anantibiotic. The concentration of epsilon polylysine in the fermentation liquor is 65.7 g / L, and the yield of epsilon polylysine reaches 80.0 g / L or above after fermentation is conducted for 240 h. Thestrain also has good passage stability, and is an epsilon PL industrial production strain with great potential.

Description

technical field [0001] The invention relates to a strain of Streptomyces albicans and its application in producing ε-polylysine, belonging to the field of microbial fermentation engineering. [0002] technical background [0003] ε-polylysine (abbreviated as ε-PL) is a homotype amino acid polymer mainly secreted by Streptomyces albulus through submerged aerobic fermentation. The degree of polymerization is generally 25-35. As a biological preservative, ε-PL has the characteristics of broad antibacterial spectrum, easy solubility in water, strong thermal stability, and high safety. It has been approved by Japan, South Korea, the United States, Europe, China and other countries and regions to be used in food and cosmetics, etc. As a biopolymer, ε-PL is widely used in biodegradable materials, drug carriers, biochip coatings, emulsifiers, high-absorbency hydrogels, and anticancer enhancers. It can be seen that ε-PL is a new type of industrial biotechnology product with excellent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P13/02C12R1/465
CPCC12P13/02C12R2001/465C12N1/205
Inventor 陈旭升毛忠贵王靓张建华张宏建
Owner JIANGNAN UNIV
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