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Epsilon-polylysine fed batch fermentation method for enhancing cell growth and bioprocess efficiency

A technology of fed-batch fermentation and polylysine, which is applied in the field of ε-polylysine fed-batch fermentation, can solve the problems that affect the synthesis ability of ε-polylysine of cells, the difficulty of fundamentally increasing cell density, and the problem of cell density. It can improve the synthesis ability of ε-polylysine, overcome the pH control fermentation, and enhance the cell activity.

Inactive Publication Date: 2013-05-01
GUANGDONG INST OF MICROORGANISM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in stage 1 and previous growth cells of this process, cells cannot synthesize ε-polylysine due to high pH, ​​and cell activity may be lost in artificially extended stage 1, which will affect the ability of cells to synthesize ε-polylysine. , due to the low pH in the synthesis stage, it is still difficult to fundamentally increase the cell density. Based on these shortcomings, it is necessary to develop a more effective fermentation process, especially a fermentation process that improves cell growth and cell activity.

Method used

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  • Epsilon-polylysine fed batch fermentation method for enhancing cell growth and bioprocess efficiency

Examples

Experimental program
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Embodiment 1

[0019] 1. Preparation of spores

[0020] Fresh epsilon-polylysine producing bacteria Streptomyces ahygroscopicus str-8 (this bacteria was preserved in China Center for Type Culture Collection on June 2, 2011, preservation address: Wuhan University, Wuhan, China, preservation The number is: CCTCC NO: M2011191, the preservation information of this strain is disclosed in the patent number: ZL201110152802.0, the name of the invention is: Streptomyces nonhygroscopicus Str-8 and the method for preparing ε-polylysine and its salt In the patent) the slant strains were streaked and inoculated in oat agar medium (ISP3) for multiple times, cultured upside down at 30°C for 7 days, washed with sterile water to collect mature spores, and prepared spore suspension as inoculum, usually The concentration of spore suspension can be obtained under the condition of about 5×10 8 CFU / mL.

[0021] Preparation of oat agar medium (ISP3): Weigh 40g of oats, wash with deionized water, place in a 1L be...

Embodiment 2

[0031] 1, spore preparation and seed liquid cultivation are the same as in Example 1, thus obtaining the seed liquid.

[0032] 2. Fed-batch fermentation process of fermenter

[0033] Take 150mL seed liquid and inoculate it in sterilized 2.85L M3G medium (5L fermenter), the temperature is 30°C, the rotation speed is 300r / min, and the ventilation is 1vvm. defoaming.

[0034]After the fermenter is inoculated with the seed liquid, when the pH of the fermentation medium in the culture system drops to about 3.6, apply 10% ammonia water on-line to control the pH of the culture system to maintain a level between 3.6 and 4.1 until the end of fermentation , that is, one-stage pH control; and when the pH drops to 3.6, the sterilized yeast powder solution (200g / L) is added continuously (adding 0.025g of yeast powder per liter of fermentation medium per hour) to Fermentation ends to enhance cell growth and activity.

[0035] During the fermentation process, cell growth and ε-polylysine ...

Embodiment 3

[0038] 1, spore preparation and seed liquid cultivation are the same as in Example 1, thus obtaining the seed liquid.

[0039] 2. Fed-batch fermentation process of fermenter

[0040] Take 150mL seed liquid and inoculate it in sterilized 2.85L M3G medium (5L fermenter), the temperature is 30°C, the rotation speed is 300r / min, and the ventilation is 1vvm. defoaming.

[0041] After the fermenter is inoculated with the seed liquid, when the pH of the fermentation medium in the culture system drops to about 4.1, apply 10% ammonia water to control the pH of the culture system on-line to maintain the pH of the culture system at 4.1 until the end of the fermentation, that is, the first stage pH control; and when the pH drops to 4.1, the yeast powder solution (200g / L) of continuous feed sterilization (adding the amount of 0.5g yeast powder in every liter of fermentation medium per hour) to the end of fermentation, to Improves cell growth and activity.

[0042] During the fermentatio...

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Abstract

The invention discloses an epsilon-polylysine fed batch fermentation method for enhancing the cell growth and the bioprocess efficiency. The method comprises the following steps: carrying out fermenting culture of epsilon-polylysine producing strains by a fermenting culture medium, supplementing 0.0.25g-0.5g of yeast powder or other organic nitrogen sources to each liter of the fermenting culture medium in a culture system each hour when the pH value of the culture system reduces to 3.6-4.1 until fermentation ending, maintaining the pH value of the culture system in a range of 3.6-4.1 by ammonia water until fermentation ending, supplementing a disinfected fed culture medium containing glucose and ammonium sulfate, and maintaining the content of glucose in the culture system in a range of 5g / L-30g / L until fermentation ending. The fast entrance of cells to the epsilon-polylysine synthesis phase avoids the cell activity loss before the cell synthesis phase and prolongs the cell epsilon-polylysine synthesis time, so the cell epsilon-polylysine synthesis capacity is substantially improved.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to an ε-polylysine fed-batch fermentation method for enhancing cell growth and biological process efficiency. Background technique: [0002] People depend on food, and food safety has become one of the hot spots at home and abroad. It is undeniable that various preservatives added to food have played an important role in food preservation and preservation, and have promoted the development of the food industry. It can be said that the modern food industry cannot do without preservatives, but the traditionally used chemically synthesized preservatives have certain potential for the human body. Hazards, such as nitrate and nitrite are carcinogenic, commonly used benzoic acid and its salts, sorbic acid and its salts are also toxic to a certain extent, in order to improve the safety of food, use natural, safe, green preservatives instead of traditional chemically synthe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/02C12R1/465
Inventor 吴清平刘盛荣莫树平柏建玲王惠惠丘明泉张菊梅
Owner GUANGDONG INST OF MICROORGANISM
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