Method for improving yield of vitamin B12 based on regulation of ammonia nitrogen index

A vitamin and B12 technology, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as premature aging of bacteria, and achieve the effect of promoting bacterial growth, increasing fermentation units, and high-density fermentation.

Pending Publication Date: 2019-09-06
HEBEI HUARONG PHARMA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problems of the prior art, the present invention provides a method for improving vitamin B vitamins based on ammonia nitrogen index regulation and control. 12 The method of the yield, the method of the present invention carries out staged feeding in batches during the fermentation process, and yeast leach juice is added in the early stage of fermentation to control the ammonia nitrogen content, and solve the problem of high concentration ammonia nitrogen in vitamin B 12 The contradiction between bacterial growth and biosynthesis during fermentation ensures the nutrients needed for bacterial growth and metabolism in the early stage of fermentation, promotes the growth of bacterial cells, avoids the phenomenon of premature aging of bacterial cells and stagnation of metabolism in the later stage, and thus increases vitamin B. 12 fermentation unit

Method used

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  • Method for improving yield of vitamin B12 based on regulation of ammonia nitrogen index

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Experimental program
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Effect test

Embodiment 1

[0020] For the yeast extract juice addition experiment, the shake flask experiment was carried out:

[0021] Carry out seed culture in a 300mL shake flask, the filling volume is 50mL, and the seed medium is: maltose 53-55m with a concentration of 20%±2%. 3 / m 3 , corn syrup 25-27Kg / m 3 , betaine 3.7-3.9Kg / m 3 , magnesium oxide 300-320g / m 3 , Ammonium sulfate 920-940g / m 3 , diammonium hydrogen phosphate 2.4-2.6g / m 3 , cobalt chloride 25-27g / m 3 , precursor 7.6-7.8g / m 3 , zinc sulfate 24-26g / m 3 , calcium carbonate 3.0-3.2Kg / m 3 . The pH of the seed medium is 7.0, the temperature is 120°C-122°C, and the sterilization time is 30min. Insert 2mL of the bacterial suspension, and shake at 29°C and 250rpm until the optical density of the bacteria in the seed liquid (OD 700 ) reached more than 0.3, transplanted.

[0022] Carry out fermentation culture in a 300mL shake flask, the filling volume is 25mL, and the fermentation medium is 1.0-1.2m maltose with a concentration of 2...

Embodiment 2

[0027] at 5m 3 Add maltose 1.0-1.2m with a concentration of 20%±2% in the fermenter 3 / m 3 , corn syrup 0.05Kg / m 3 , Betaine 11-13Kg / m 3 , magnesium oxide 600-620g / m 3 , ammonium sulfate 1.1-1.3Kg / m 3 , diammonium hydrogen phosphate 600-620g / m 3 , cobalt chloride 90-92g / m 3 , precursor 65-67g / m 3 , zinc sulfate 24-26g / m 3 , urea 450-470g / m 3 , ferric chloride 0.6-0.7Kg / m 3 , calcium carbonate 1.1-1.3Kg / m 3 . Adjust the pH value of the fermentation medium to 7.00. After the batching is completed, steam sterilization is carried out. Sterilization conditions: pressure 0.10-0.13MPa, temperature 120°C-122°C, sterilization time 30min, after sterilization is completed, cool and pressurize with sterile air , awaiting transplantation.

[0028] The cultivated seed solution is transferred into the fermentation medium for cultivation, and the inoculation amount is controlled at 10%. Culture conditions: tank pressure 0.04-0.05MPa, tank temperature 30°C-33°C, air flow 15-20m ...

Embodiment 3

[0032] at 5m 3 Add maltose 1.0-1.2m with a concentration of 20%±2% in the fermenter 3 / m 3 , corn syrup 0.05Kg / m 3 , Betaine 11-13Kg / m 3 , magnesium oxide 600-620g / m 3 , ammonium sulfate 1.1-1.3Kg / m 3 , diammonium hydrogen phosphate 600-620g / m 3 , cobalt chloride 90-92g / m 3 , precursor 65-67g / m 3 , zinc sulfate 24-26g / m 3 , urea 450-470g / m 3 , ferric chloride 0.6-0.7Kg / m 3 , calcium carbonate 1.1-1.3Kg / m 3 . Adjust the pH value of the fermentation medium to 7.00. After the batching is completed, steam sterilization is carried out. Sterilization conditions: pressure 0.10-0.13MPa, temperature 120°C-122°C, sterilization time 30min, after sterilization is completed, cool and pressurize with sterile air , awaiting transplantation.

[0033] The cultivated seed solution is transferred into the fermentation medium for cultivation, and the inoculation amount is controlled at 10%. Culture conditions: tank pressure 0.04-0.05MPa, tank temperature 30°C-33°C, air flow 15-20m ...

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Abstract

Provided is a method for improving the yield of vitamin B12 based on regulation of an ammonia nitrogen index. In the fermentation process of vitamin B12, stage-by-stage batch feeding is carried out; in the earlier stage of fermentation, a yeast extract juice is refilled, so a certain content of ammonia nitrogen is maintained at a certain stage, so as to ensure the nitrogen source needed for growthand metabolism of bacteria. The stage-by-stage batch feeding is carried out in the fermentation process, the yeast extract juice is refilled in the earlier stage of fermentation, the concentration ofammonia nitrogen is controlled, the contradiction between the growth of the bacteria and biosynthesis of high-concentration ammonia nitrogen in the fermentation of vitamin B12 is solved, the nutritional components required for the growth and metabolism of the bacteria are ensured in the early stage of fermentation, the growth of bacteria is promoted, the phenomena of premature senescence and metabolic stagnation in the later period are avoided, and the fermentation unit of vitamin B12 is increased.

Description

technical field [0001] The invention relates to a method for improving vitamin B 12 The method of yield, especially relates to a kind of regulation and control based on ammonia nitrogen index to improve vitamin B 12 yield method. Background technique [0002] B Vitamins in Nature 12 All are synthesized by microorganisms, and higher animals and plants cannot produce vitamin B 12 . Vitamin B 12 Obtained through strain cultivation and fermentation. Vitamin B 12 Its main physiological function is to participate in the manufacture of bone marrow red blood cells, prevent pernicious anemia, and prevent brain nerves from being damaged. [0003] Vitamin B present 12 The production of aerobic sword bacteria (Ensifer adhaerens) is obtained through fermentation and purification. The main components of the fermentation medium include: organic nitrogen source, carbon source, betaine, precursors and some inorganic salts and trace elements. The main function of the nitrogen source ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/42C12R1/01
CPCC12P19/42
Inventor 王俊刚王继红刘汉忠曹艳霞穆启恒李英然李雪然张敬坤王红袁玉伟
Owner HEBEI HUARONG PHARMA
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