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Epsilon-polylysine high-yielding strain and epsilon-polylysine production method

A technology of polylysine and high-yielding strains, applied in the biological field, can solve the problems of large equipment investment, complex separation process and high cost, and achieve the effects of small equipment investment, low energy consumption and improved product yield

Active Publication Date: 2017-02-22
SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, there are two main problems that plague the large-scale industrialization of ε-polylysine: one is the limited acid production ability of existing strains; the other is the complicated separation process, which generally requires sterilization, ion exchange, decolorization, concentration, Steps such as drying; such as a method for extracting ε-polylysine and its salt from fermentation broth provided by Chinese patent 200910152931.2, the process steps are complicated, the equipment investment is large, the cost is high, and a large amount of sewage will be generated during the separation process

Method used

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  • Epsilon-polylysine high-yielding strain and epsilon-polylysine production method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Screening of high-yield strain EA-19

[0029] (1) The spores of Streptomyces albicans NBRC14147 were made into a spore suspension using tirs-maleic acid buffer, and the spore concentration was about 10 7 ~10 8 individual / mL.

[0030] (2) Take 5 mL of the spore suspension and add it to a Φ9 cm sterile plate; place the sterile plate in a UV mutagenesis box at a distance of 30 cm from a 15 W UV lamp for 30 s of ultraviolet irradiation mutagenesis.

[0031] (3) Take the sterile plate out of the UV mutagenesis chamber, then add nitrosoguanidine to the spore suspension in the sterile plate so that the final concentration of nitrosoguanidine is 0.8 mg / mL, and shake for 30 min in the dark ; terminate mutagenesis.

[0032] (4) After the mutagenesis is over, wash the cells and add 5 mL of dark culture overnight (the culture time is about 16 hours). Fermentation medium: glucose 50 g / L, yeast powder 5 g / L, ammonium sulfate 10 g / L, ferrous sulfate 0.03 g / L, zinc sulfat...

Embodiment 2

[0049] Example 2 High-yield strain EA-19 Fermentation Separation Coupling Production ε-Polylysine

[0050] (1) Inoculate the EA-19 spore suspension prepared in Example 1 into the liquid seed medium at a 2% inoculum amount, and incubate at 30 °C and 250 r / min for 24 h; obtain the EA-19 seed liquid. The composition of the seed medium is glycerol 20 g / L, yeast powder 5 g / L, ferrous sulfate 0.03 g / L, zinc sulfate 0.04 g / L, magnesium sulfate 0.5 g / L, dipotassium hydrogen phosphate 0.8 g / L, phosphoric acid Potassium dihydrogen 1.36g / L.

[0051] (2) Inoculate the EA-19 seed liquid into the fermentation medium according to the inoculum amount of 5%, the culture temperature is 30 ℃, the stirring speed is 400 200-500r / min, the ventilation ratio is 1:1 (V / V m), and the fermentation starts to cultivate. The fermentation medium is glucose 50 g / L, yeast powder 5 g / L, ammonium sulfate 10 g / L, ferrous sulfate 0.03 g / L, zinc sulfate 0.04 g / L, magnesium sulfate 0.5 g / L, dipotassium hydrogen p...

Embodiment 3

[0058] Example 3 High-yield strain EA-19 Fermentation separation coupled production of ε-polylysine

[0059] (1) Inoculate the EA-19 spore suspension prepared in Example 1 into the liquid seed medium at a 2% inoculation amount, and incubate at 30 °C and 250 r / min for 24 h; the EA-19 seed liquid was obtained. The composition of the seed medium is glycerol 20 g / L, yeast powder 5 g / L, ferrous sulfate 0.03 g / L, zinc sulfate 0.04 g / L, magnesium sulfate 0.5 g / L, dipotassium hydrogen phosphate 0.8 g / L, phosphoric acid Potassium dihydrogen 1.36g / L.

[0060] (2) Inoculate the obtained EA-19 seed liquid into the fermentation medium according to the inoculum amount of 5%. The culture temperature is 30 ℃, the stirring speed is 400 r / min, the ventilation ratio is 1:1 V / V·m, and the fermentation culture is started. The fermentation medium is glucose 50 g / L, yeast powder 5 g / L, ammonium sulfate 10 g / L, ferrous sulfate 0.03 g / L, zinc sulfate 0.04 g / L, magnesium sulfate 0.5 g / L, dipotassium h...

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Abstract

The invention relates to an epsilon-polylysine high-yielding strain and an epsilon-polylysine production method, and belongs to the biotechnical field. The epsilon-polylysine high-yielding strain EA-19 is named as Streptomyces albus, and the preservation number is CGMCC No.10156. The strain EA-19 is used to carry out fed batch fermentation culture, and the concentration of epsilon-polylysine in a fermentation liquid reaches 38.5-39 g / L. The epsilon-polylysine production method comprises is characterized in that the strain EA-19 is adopted to carry out fermentation culture. A bubbling technology is adopted, ventilation is carried out in and after the fermentation culture process, so epsilon-polylysine generated after fermentation exists in foams, and is separated from the fermentation liquid, so the technical problem of difficulty separation of epsilon-polylysine in present preparation methods is solved, a step of ion exchange which causes generation of a large amount of wastewater is omitted, and the product yield is improved by 50% or above. The method has the advantages of simple process, easiness in amplification, low energy consumption, small device investment, and easiness in realization of large-scale production.

Description

technical field [0001] The invention relates to a high-yielding ε-polylysine strain and a method for producing ε-polylysine, belonging to the field of biotechnology. Background technique [0002] ε-polylysine (ε-polylysine) belongs to microbial food preservatives, and is a new type of high-efficiency and broad-spectrum biological preservatives. ε-polylysine is a polymer of L-lysine linked by α-carboxyl and ε-amino groups, containing 25~35 lysine residues, the finished product is light yellow powder, easily soluble in water, hygroscopic Strong, slightly bitter. Compared with other preservatives, ε-polylysine has the characteristics of broad antibacterial spectrum, high antibacterial efficiency, good stability and high safety. ε-polylysine has antibacterial effects on Gram-positive bacteria, Gram-negative bacteria, yeasts, and molds, and also has inhibitory effects on heat-resistant Bacillus and some viruses; the amount added to food is very small; it can It adapts to a wid...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/01C12P13/02C12R1/47
CPCC12N15/01C12P13/02C12N1/205C12R2001/47
Inventor 刘飞张林军朱希强张金华侯重文袁超王林张祥奎凌沛学
Owner SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
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