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Method for effectively producing vitamin K2 by improving cell membrane permeability

A vitamin and permeability technology, applied in the field of biotechnology preparation, can solve problems such as unsatisfactory vitamin K2 yield, and achieve the effects of easy industrial application, increased concentration, and economical fermentation process

Active Publication Date: 2017-12-15
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

At present, the strains used for synthesizing vitamin K2 in the field of fermentation include Gram-negative bacteria Flavobacterium (mainly producing MK-4) and Gram-positive bacteria Bacillus natto (mainly producing MK-6), but the vitamin K2 output is low. not ideal

Method used

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  • Method for effectively producing vitamin K2 by improving cell membrane permeability
  • Method for effectively producing vitamin K2 by improving cell membrane permeability
  • Method for effectively producing vitamin K2 by improving cell membrane permeability

Examples

Experimental program
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Effect test

Embodiment 1

[0040] Such as figure 1 As shown, a method of effectively producing vitamin K2 by improving cell membrane permeability of the present embodiment comprises the following steps:

[0041] (1) Strain activation

[0042] Using Flavobacterium (provided by China Common Microorganism Culture Collection Center, No. CCTCC AB 2010137) as the starting bacterium, inoculate the Flavobacterium in the slant medium for activation, and the composition of the slant medium is: glycerol 5g / L, glucose 10g / L , yeast extract 2.0g / L, fish powder peptone 5.0g / L, sodium chloride 5.0g / L, agar 1.5%, pH7.4, culture temperature is 30℃, culture for 48h;

[0043] (2) Primary seed preparation

[0044] The activated Flavobacterium strains were inoculated in the seed medium added with 0.01g / L Span and cultivated for 12h to make first-grade seeds for later use; the components of the seed medium were: glycerol 5g / L, glucose 10g / L, yeast extract 2.0g / L, peptone 5.0g / L, sodium chloride 5.0g / L, Span 0.01g / L, pH 7....

Embodiment 2

[0051] Such as figure 1 As shown, a method of effectively producing vitamin K2 by improving cell membrane permeability of the present embodiment comprises the following steps:

[0052] (1) Using Flavobacterium (provided by the China Common Microorganism Culture Collection Center, No. CCTCC AB 2010137) as the starting bacterium, the Flavobacterium was inoculated in the slant medium for activation. The slant medium composition was: glycerol 5g / L, glucose 10g / L, yeast extract 2.0g / L, fish powder peptone 5.0g / L, sodium chloride 5.0g / L, agar 1.5%, pH7.4, culture temperature is 30℃, culture for 48h;

[0053] (2) Inoculate the activated strains of the slant medium into the seed medium supplemented with 0.05g / L OP-10, the composition of the seed medium is: glycerol 5g / L, glucose 10g / L, yeast extract 2.0g / L, peptone 5.0g / L, sodium chloride 5.0g / L, OP-10 0.05g / L, pH7.2, culture temperature 37℃, culture 18h;

[0054] (3) Inoculate the obtained primary seed culture solution into the se...

Embodiment 3

[0058] Such as figure 1 As shown, a method of effectively producing vitamin K2 by improving cell membrane permeability of the present embodiment comprises the following steps:

[0059] (1) Using Flavobacterium (provided by the China Common Microorganism Culture Collection Center, No. CCTCC AB 2010137) as the starting bacterium, the Flavobacterium was inoculated in the slant medium for activation. The slant medium composition was: glycerol 5g / L, glucose 10g / L, yeast extract 2.0g / L, fish powder peptone 5.0g / L, sodium chloride 5.0g / L, agar 1.5%, pH7.4, culture temperature is 30℃, culture for 48h;

[0060] (2) Inoculate the Flavobacterium strains after the activation of the slant medium into the seed medium added with 1g / L OP-10, the composition of the seed medium is: glycerol 5g / L, glucose 10g / L, yeast extract 2.0 g / L, peptone 5.0g / L, sodium chloride 5.0g / L, OP-10 1g / L, pH 7.2, culture temperature 37℃, culture 24h;

[0061] (3) Inoculate the primary seed culture solution obtain...

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Abstract

The invention discloses a method for effectively producing vitamin K2 by improving cell membrane permeability. The method comprises the following steps: (1) inoculating a vitamin K2 production strain into a bevel culture medium and activating; (2) inoculating the activated strain into a seed medium containing a nonionic surfactant for culturing; (3) continuously inoculating the acquired culture liquid into the seed medium containing a nonionic surfactant for culturing; and (4) inoculating the culture liquid into a fermentation medium, culturing for 144-240h at 100-700rpm and 0.5-5vvm, fermenting for 0-72h and then adding the nonionic surfactant, performing plug-in low-power ultrasonic separating treatment, sampling at fixed time and detecting the cell and vitamin K2 concentration. The method disclosed by the invention has the advantages that a simple, economical and effective fermenting technology is supplied, the vitamin K2 concentration is effectively increased, the production efficiency is increased and the production cost is lowered.

Description

technical field [0001] The invention relates to the field of biotechnology preparation, in particular to a method for effectively producing vitamin K2 by improving cell membrane permeability. Background technique [0002] Vitamin K2 (menaquinone, abbreviated as MK) is a fat-soluble vitamin and one of the indispensable important vitamins in the human body. It can affect the synthesis and coagulation of prothrombin, and has the characteristics of easy absorption, high bioavailability and strong biological activity Moreover, it has many advantages, such as little toxic and side effects on the human body, and is currently clinically used for the treatment of vitamin K deficiency hemorrhage, prevention and treatment of osteoporosis, and anti-tumor. [0003] At present, the production methods of vitamin K2 mainly focus on chemical synthesis and natural product extraction, and the production of vitamin K2 by biological methods is still in its infancy. The advantages of microbial p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P7/66C12N1/20C12R1/07C12R1/20
CPCC12N1/20C12P7/66
Inventor 郑之明杨强赵根海刘会王鹏王丽方雪孙小雯倪文枫
Owner HEFEI INSTITUTES OF PHYSICAL SCIENCE - CHINESE ACAD OF SCI
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