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Method for performing high-density fermentation on bacillus aceticus

A technology of high-density fermentation and acetic acid bacteria, which is applied in the field of high-density fermentation of acetic acid bacteria, can solve the problems of long cultivation period and achieve the effect of shortening the production period

Inactive Publication Date: 2018-05-29
SOUTH CHINA INST OF COLLABORATIVE INNOVATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The Chinese patent application with the publication number CN103820368A discloses "Preparation method and application of acetic acid bacteria freeze-dried bacteria powder", adopting 5-25 wt% sodium hydroxide aqueous solution to maintain the pH of the fermentation medium, but the culture period is longer, requiring 20-20 wt% 30h

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  • Method for performing high-density fermentation on bacillus aceticus

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Comparison scheme
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Embodiment 1

[0021] A high-density fermentation method for acetic acid bacteria, comprising the steps of:

[0022] (1) Activation of strains: pick well-growing Acetobacter seeds from the preservation medium and inoculate them on the agar medium, and cultivate them at 30°C for 24 hours to obtain activated strains; the composition of the agar medium is: yeast extract 1 %, glucose 1%, agar 2%, ethanol 2%, pH 5.5-6.0, percentages are mass percentages, and the balance is water;

[0023] (2) Shake flask culture: Inoculate the activated bacteria in step (1) into the shake flask, shake and cultivate for 18 hours, and obtain the seed liquid; shake flask medium formula: yeast extract 1%, glucose 1%, KH 2 P0 4 0.25%, ethanol 2%, pH 5.5-6.0, the percentages are mass percentages, and the balance is water; the liquid volume in a 250mL shaker flask is 50mL; culture conditions: shaker flask speed 150rpm, temperature 30°C;

[0024] (3) Seed tank expansion: the liquid cultured in the shake flask is used ...

Embodiment 2

[0027] A high-density fermentation method for acetic acid bacteria, comprising the steps of:

[0028] (1) strain activation and shake flask culture are the same as in Example 1

[0029] (2) Seed tank expansion: the liquid cultivated by the shake flask is inserted into a 10L fermenter with an inoculum size of 10% as the seed liquid and cultivated; the liquid volume of the 10L fermenter is 60%; the composition of the culture medium: yeast extract 4%, Glucose 5%, KH 2 P0 4 0.75%, ethanol 2%, pH 5.5-6.0; culture conditions: stirring speed 300rpm, ventilation volume 1vvm, temperature 30°C, adding 5M sodium hydroxide, controlling the pH of the fermentation broth to 5.5-6.0, culture time 10h;

[0030] (3) Fermentation tank fermentation: Transplant by pressure difference, the seed liquid in the seed tank is inserted in the fermentation tank by 10% inoculum amount and fermented; 100L fermentation tank liquid volume 70%; Culture medium composition: yeast paste 4 %, Glucose 5%, KH 2...

Embodiment 3

[0032] A high-density fermentation method for acetic acid bacteria, comprising the steps of:

[0033] (1) strain activation and shake flask cultivation are the same as in Example 1, and the seed tank expansion is the same as in Example 2;

[0034] (2) Fermentation tank fermentation: Transplant by pressure difference, the seed liquid in the seed tank is inserted in the fermentation tank by 10% inoculum amount and fermented; 100L fermentation tank liquid volume 70%; Culture medium composition: yeast extract 4 %, Glucose 5%, KH 2 P0 4 0.75%, ethanol 2%, pH 5.5-6.0; culture conditions: stirring speed 300rpm, ventilation 1vvm, temperature 30°C, adding 5M sodium hydroxide, controlling the pH of the fermentation broth to 5.5-6.0, and adding light at the same time Density OD 600 =2 cell slurry and 5g / L bitter gourd polypeptide (the addition amount of these two substances is 300mL respectively), and the culture time is 12h. Described cell slurry refers to fermented liquid (number ...

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Abstract

The invention belongs to the technical field of food microorganisms and discloses a method for performing high-density fermentation on bacillus aceticus. The method comprises the following steps: (1)performing bacterium activation; (2) performing shake-flask culture; (3) performing culture amplification of seed tanks; and (4) performing fermentation in fermentation tanks, namely inoculating a seed liquid obtained in the step (3) into culture mediums in the fermentation tanks according to an inoculation amount of 1-20%, performing culture under culture conditions that the stirring rotation speed is 100-300 rpm, the air introduction amount is 0.1-1.0 vvm, and the temperature is 28-30 DEG C, feeding 5-10 M of sodium hydroxide, controlling the pH value of a fermentation liquid to 5.5-6.0, feeding cell pulp and polypeptide-k, and culturing for 8-12 hours, wherein the cell pulp is cell pulp of bacillus aceticus. By adopting the method, the production cycle is greatly shortened, bacteria canbe relatively greatly enriched, and high-density fermentation of 1.24-3.29*10<9>cfu / mL is achieved.

Description

technical field [0001] The invention relates to a method for high-density fermentation of acetic acid bacteria, which belongs to the field of food biotechnology. Background technique [0002] Acetobacter, Gram-negative, does not produce spores, can be used to make vinegar, aerobic respiration, and can oxidize ethanol (alcohol) into acetic acid under aerobic, neutral and acidic conditions. The content of alcohol and base acid in the fermentation medium had a significant effect on the growth and acid production of bacteria in acetic acid fermentation. Although Acetobacter has a certain tolerance to acetic acid, as the fermentation time prolongs, the acid production gradually increases, and Acetobacter may be killed by the acid produced by itself. [0003] Compared with the rapid development of the dairy industry, the research on Acetobacter inoculum in my country is relatively backward. The traditional fermentation process often adopts batch fermentation and single-batch fed-...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/02
CPCC12N1/20
Inventor 刘艺婷杨继国任杰刘玮
Owner SOUTH CHINA INST OF COLLABORATIVE INNOVATION
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