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Method for promoting pseudomonas denitrificans to generate vitamin B12

A technology for Pseudomonas and vitamins, which is applied in the field of improving the production of vitamin B12 by Pseudomonas denitrification, can solve the problems of premature aging of the bacteria, affect the production of vitamin B12, etc. cost effect

Inactive Publication Date: 2019-06-04
山东泓达生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The control conditions of the aerobic fermentation of Pseudomonas denitrifica to produce vitamin B12 are relatively strict. Once the control is not good, it will lead to premature aging of the bacteria and affect the production of vitamin B12

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria

[0017] A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria comprises the following steps:

[0018] S1. Mother bottle culture process control: Scrape an appropriate amount of activated Pseudomonas denitrification second-generation slant and inoculate it into the mother bottle culture medium. The temperature is controlled at 27°C, the humidity is controlled at 30-35%, and the culture time is 18 hours; at this time, the mother bottle medium OD: 0.220; pH: 7.34;

[0019] S2. Shake flask culture process control: Inject the standard Pseudomonas denitrification mother bottle culture medium (no miscellaneous bacteria, OD value 0.220) into the shake flask culture medium under aseptic conditions, and the inoculum amount is 10%. The temperature of the shaker in the early stage of fermentation (0-35h) is controlled at 27±0.5°C; the tempe...

Embodiment 2

[0025] Example 2 A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria

[0026] A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria comprises the following steps:

[0027] S1. Mother bottle culture process control: Scrape an appropriate amount of activated Pseudomonas denitrification second-generation slant and inoculate it into the mother bottle culture medium. The temperature is controlled at 29°C, the humidity is controlled at 50-60%, and the culture time is 24 hours; at this time, the mother bottle medium OD: 0.212; pH: 7.42;

[0028] S2. Shake flask culture process control: Inject the standard Pseudomonas denitrification mother bottle culture medium (no miscellaneous bacteria, OD value 0.212) into the shake flask culture medium under aseptic conditions, the inoculum volume is 10%, The temperature of the shaker in the early stage of fermentation (0-38h) is controlled at 29±0.5°C; the temperatu...

Embodiment 3

[0034] Example 3 A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria

[0035] A method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria comprises the following steps:

[0036] S1. Mother bottle culture process control: Scrape an appropriate amount of activated Pseudomonas denitrification second-generation slant and inoculate it into the mother bottle culture medium. The temperature is controlled at 28°C, the humidity is controlled at 10-50%, and the culture time is 22 hours; at this time, the mother bottle medium OD: 0.207; pH: 7.39;

[0037] S2. Shake flask culture process control: Inject the standard Pseudomonas denitrification mother bottle culture medium (no miscellaneous bacteria, OD value 0.207) into the shake flask culture medium under aseptic conditions, and the inoculum amount is 10%. The temperature of the shaker in the early stage of fermentation (0-39h) is controlled at 28±0.5°C; the tempe...

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Abstract

The invention relates to the technical field of biology fermentation, in particular to a method for promoting pseudomonas denitrificans to generate vitamin B12. The method for promoting pseudomonas denitrificans to generate vitamin B12 comprises the following steps of through a mother bottle culture medium, a shake flask culture medium and a supplemented medium optimized through massive experiments, under the optimized fermentation condition, production bacterial strains of vitamin B12 are cultured. In 0-39h of the front period of the fermentation process, the temperature is controlled to be at a relatively-low level, namely 26-30 DEG C, the strains are controlled to be at appropriate growth rate and favorable mushroom shapes are maintained. In the middle and later periods of the fermentation process, the temperature of the fermentation system is increased and maintained to be at a relatively-low level, namely 30-34 DEG C, the growth of thalli is promoted, starting and accumulation ofsynthesis of the vitamin B12 are promoted, and further, after the fermentation is finished, the fermentation unit of the vitamin B12 is as high as 198mg / L, so that the yield and the production efficiency of the vitamin B12 are increased, the energy consumption is reduced, and the cost is reduced.

Description

technical field [0001] The invention relates to the technical field of biological fermentation, in particular to a method for improving the production of vitamin B12 by Pseudomonas denitrification bacteria. Background technique [0002] Vitamin B12 (Vitamin B12), also known as cobalamin, is a general term for a class of corrin compounds containing cobalt. It was first discovered in 1926 as an "anti-pernicious anemia factor". Vitamin B12 is an important biologically active substance and also a growth factor for many microorganisms and animals, and is widely used in medicine, food and animal husbandry. Due to the extremely complex molecular structure of vitamin B12, its total chemical synthesis requires more than 70 reaction steps and is expensive, so vitamin B12 is almost always produced through microbial fermentation. In nature, there are two different pathways for the biosynthesis of vitamin B12, namely the aerobic pathway and the anaerobic pathway, but the biosynthesis of...

Claims

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Application Information

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IPC IPC(8): C12P19/42C12N1/20C12R1/38
Inventor 刘瑞艳马杰希刘伟张彩霞寇凤雨
Owner 山东泓达生物科技有限公司
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