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A method for improving the production of vitamin K2 (mk) synthesized by Flavobacterium by adding inducing substances

An inducing substance, the technology of Flavobacterium, which is applied in the field of microbial fermentation, can solve the problem of low yield of MK in fermentation production, and achieve the effect of increasing the dry weight ratio of bacteria, simple operation, and increased yield

Active Publication Date: 2020-02-18
HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention will provide a method for improving the production of MK synthesized by Flavobacterium by adding inducing substances, so as to overcome the low yield of MK produced by fermentation of Flavobacterium in the prior art

Method used

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  • A method for improving the production of vitamin K2 (mk) synthesized by Flavobacterium by adding inducing substances
  • A method for improving the production of vitamin K2 (mk) synthesized by Flavobacterium by adding inducing substances
  • A method for improving the production of vitamin K2 (mk) synthesized by Flavobacterium by adding inducing substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Activation of Flavobacterium strains: Pick a ring of bacteria solution from a frozen glycerol tube and streak it on a plate containing solid medium by the streak dilution method, and culture it upside down at 25°C for 24 hours. Inoculate the colony on a fresh slant containing solid medium, and continue to maintain the temperature and invert the culture for 20 hours. The components of the above solid medium include 10g / L maltose, 10g / L corn steep liquor, 0.5g / L peptone, 0.2g / L yeast extract , 0.5g / L K 2 HPO 4 , 0.5g / L NaCl, 0.1g / L MgSO 4 ·7H 2 O and 15g / L agar powder.

[0031] (2) Seed cultivation of Flavobacterium strains: Pick a mature slant and use an inoculation loop to scrape a ring of bacterial lawn and inoculate it into a 500ml Erlenmeyer flask containing 50ml of seed medium, and culture at 25°C and 100rpm for 20h to obtain a seed solution . Seed medium components include 10g / L maltose, 10g / L corn steep liquor, 0.5g / L peptone, 0.2g / L yeast extract, 0.5g / L...

Embodiment 2

[0037] (1) Activation of Flavobacterium strain strains: pick a ring of bacterial liquid from the frozen glycerin tube and apply it on a plate containing solid medium by the dilution plating method, preferably adopt the dilution plating method to apply the bacterial liquid to the plate, Inverted culture at 37°C for 48 hours, pick a single colony and inoculate it on a fresh slant containing solid medium, and continue to maintain the temperature for 30 hours. The composition of the solid medium includes 40g / L maltose, 40g / L corn steep liquor, 10g / L L peptone, 2g / L yeast extract, 5g / L K 2 HPO 4 , 5g / LNaCl, 0.5g / LMgSO 4 ·7H 2 O and 20g / L agar powder.

[0038] (2) Seed cultivation of Flavobacterium strains: Pick a mature slant and use an inoculation loop to scrape a ring of bacterial lawn and inoculate it into a 500ml Erlenmeyer flask containing 100ml of seed medium, and culture at 37°C and 200rpm for 30h to obtain a seed solution , the ingredients of the above seed medium inclu...

Embodiment 3

[0043] (1) Activation of Flavobacterium strains: pick a ring of bacterial liquid from the frozen glycerol tube on a plate containing solid medium, preferably use the dilution plating method to spread the bacterial liquid on the plate, and incubate it upside down at 37°C for 48 hours , pick a single colony and inoculate it on a fresh slope containing a solid medium, and continue to maintain the temperature and invert it for 30 hours. The components of the solid medium include 40g / L maltose, 40g / L corn steep liquor, 10g / L peptone, 2g / L yeast Extract, 5g / LK 2 HPO 4 , 5g / LNaCl, 0.5g / LMgSO 4 ·7H 2 O and 20g / L agar powder.

[0044] (2) Seed cultivation of Flavobacterium strains: Pick a mature slant and use an inoculation loop to scrape a ring of bacterial lawn and inoculate it into a 500ml Erlenmeyer flask containing 100ml of seed medium, and culture at 37°C and 200rpm for 30h to obtain a seed solution , the ingredients of the above seed medium include 40g / L maltose, 40g / L corn ...

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Abstract

The invention relates to a method for increasing the yield of flavobacterium synthetic vitamin K2 by means of adding inductive substances. The method specifically includes the steps of flavobacterium strain activation, flavobacterium seed culture, flavobacterium fermentation culture, centrifuging of fermentation liquor, freeze drying, methyl alcohol treatment and the like. Precursor substances, namely, 1,4-dyhydroxy-2-naphthoic acid and precursor analogues, namely, VK3 or 3-methyl-2-buten-1-ol (YWXC) are added to a fermentation medium to serve as the inductive substances, biosynthesis of the MK is promoted, and obtained leaching liquor is detected. The result shows that compared with a control group, the yield of the flavobacterium synthetic vitamin K2 to which the inductive substances are added is remarkably increased, and the dry weight ratio of thalli is also greatly increased through the yield of the vitamin K2. According to the method, operation is simple, controllability is high, the result is stable, and safety and environment friendliness are achieved. In addition, the yield of flavobacterium synthetic MK is greatly increased by adding the inductive substances, and a novel thought and means are provided for industrial large-scale production of the vitamin MK.

Description

technical field [0001] The invention belongs to the microbial fermentation industry, and in particular relates to a method for increasing the yield of MK fermented by Flavobacterium by adding inducing substances. Background technique [0002] Vitamin K (Vitamin K, VK) is a general term for a group of substances containing a common chemical structure "2-methyl-1,4-naphthoquinone ring". According to the different chemical structures of the side chains connected to the 3' position of the naphthoquinone ring, VK can be divided into different categories. There are two main types of naturally occurring VK, VK 1 (Phylloquinore, PK, phylloquinones) and VK 2 (Menaquinone, MK, menaquinones), both of which are fat-soluble vitamins, and can be synthesized by microorganisms. VK 1 Mainly produced by plants, VK 2 Mainly produced by microorganisms. And synthetic VK has VK 3 、VK 4 、VK 5 Wait. Among them, VK is biologically active in animals 2 , while VK 1 and VK 3 Both need to ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/66C12R1/20
Inventor 郑之明吴荷芳王鹏王丽赵根海刘会王晗孙孝娟孙小雯尉鸿飞李哲敏方雪
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI