Streptomyces albulus (GS-114) and method of the same to prepare epsilon-polylysine

A technology of Streptomyces parvus and microbial strains, applied in the field of Streptomyces and its preparation of ε-polylysine, can solve the problems of not being suitable for large-scale application in breeding work, general screening effect, and high price, and achieve The effect of excellent passage stability

Inactive Publication Date: 2020-02-18
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 albulus (GS-114) and method of the same to prepare epsilon-polylysine
  • Streptomyces albulus (GS-114) and method of the same to prepare epsilon-polylysine
  • Streptomyces albulus (GS-114) and method of the same to prepare epsilon-polylysine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Embodiment 1: the breeding of Streptomyces parvus GS-114

[0050] 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.

[0051] 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.

[0052] Specific steps are as follows:

[0053] (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 Bet...

Embodiment 2

[0055] Example 2: Morphological characteristics of the starting bacterium M-Z18 and the high-yield bacterium GS-114

[0056] The morphology of the strains was observed with naked eyes and microscopic analysis.

[0057] 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.

[0058] The colony of strain GS-114 is 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 the YP liquid medium, the hyphae of GS-114 were loose and the balls were small.

Embodiment 3

[0059] Example 3: The difference in the shake flask output of high-yielding bacteria GS-114 and starting bacteria M-Z18 under different medium conditions

[0060] M3G seed medium (by g / 100mL): 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.

[0061] RSM liquid fermentation medium (by g / 100mL): glucose 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, sterilized at 115°C for 20 minutes, fermented at natural pH.

[0062] YP liquid fermentation medium (by g / 100mL): glucose 3, (NH 4 ) 2 SO 4 0.5, yeast powder 0.8, KH 2 PO 4 0.4, MgSO 4 0.08, FeSO 4 0.005, sterilized at 115°C for 20 minutes, fermented at natural pH.

[0063] The purified GS-114 spores were inoculated into the medium filled with M3G, and cultured in shake flasks at 30°C×200rpm for 24h to prepare seed liquid. The seed so...

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Abstract

The invention discloses streptomyces albulus (GS-114) and a method of the same to prepare epsilon-polylysine, and belongs to the field of microbial fermentation engineering. The provided streptomycesalbulus (GS-114) has resistance to streptomycin, gentamicin and rifamycin with 3 [mu]g / mL, 1 [mu]g / mL and 0.1 [mu]g / mL or higher concentrations, and can efficiently synthesize epsilon-PL in quantity,the yield with 192 h of fermentation can reach 56.3 g / L, and production efficiency can reach up to 7.03 g / L / d. The streptomyces albulus (GS-114) also has good passage stability and has strong potential for the industrial production of the epsilon-PL.

Description

technical field [0001] The invention relates to a strain of streptomyces and a method for preparing ε-polylysine, belonging to the field of microbial fermentation engineering. [0002] technical background [0003] ε-polylysine (abbreviated ε-PL) is a new type of natural biological food preservative mainly secreted by Streptomyces albulus through liquid aerobic fermentation. ε-PL has a wide antibacterial spectrum, is easily soluble in water, has strong thermal stability and high safety. It has been successively approved as a new food additive by Japan, the United States, South Korea, Europe and China and is widely used in the food industry. At the same time, as a cationic biopolymer, ε-PL is widely used in biodegradable materials, drug carriers, biochip coatings, emulsifiers, high-absorbency hydrogels and anti-cancer enhancers, etc., and has a broad market Application prospect. At present, ε-PL is relatively expensive in the domestic and international markets (about 1200 yu...

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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