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Menadione-7(MK-7) high-yielding strain and application thereof

A technology of menadione and high-yield strains, which is applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., and can solve the problems of low fermentation yield, long fermentation cycle, and high cost of raw materials

Inactive Publication Date: 2014-06-18
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, from the current point of view, MK-7 has not been able to achieve industrialized fermentation production. The main reason is that the fermentation method has a low yield, a long fermentation cycle, and high raw material costs. In view of this, it is necessary to carry out MK-7 production strains. screening optimization to address the above issues

Method used

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  • Menadione-7(MK-7) high-yielding strain and application thereof
  • Menadione-7(MK-7) high-yielding strain and application thereof
  • Menadione-7(MK-7) high-yielding strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Screening and identification of Bacillus amyloliquefaciens subspecies Y-2

[0021] 1. the screening method of Bacillus amyloliquefaciens subspecies Y-2 provided by the invention is:

[0022] Take 4 natto samples (about 0.5g) in 10mL sterile saline, shake the water bath at 80°C for 10min. Take 100 μL of the extract, dilute it with an appropriate amount of sterile saline, and finally take 200 μL of the diluted solution and spread it on the LBG solid medium, culture it overnight at 37°C, isolate a single colony, and select the colony that can draw silk from it, and transfer it to the LBG liquid In the culture medium, culture at 37°C for 24 hours, detect the MK-7 content therein, and select the bacterium with the highest MK-7 production, and name it Y-2.

[0023] 2. MK-7 provided by the present invention extracts and detects method as follows:

[0024] Centrifuge the fermented liquid after fermentation at 7000r / min for 5min, separate the supernatant from the bac...

Embodiment 2

[0032] The production method of corn flour liquefaction liquid: mix industrial corn flour with water at a ratio of 30% (mass volume ratio w / v), then gelatinize in a boiling water bath, and then add high-temperature amylase at a ratio of 0.1% to liquefy in a boiling water bath for 20 minutes. Afterwards, filter with 4 layers of gauze to obtain the supernatant, and then replenish to the original volume with water to obtain the required cornflour liquefaction solution, in which the reducing sugar content is about 60g / L.

[0033] Fermentation medium: corn flour liquefaction 1L (reducing sugar content is about 60g / L), soybean meal powder 40g / L, soybean peptone 15g / L, yeast extract 15g / L, K 2 HPO 4 0.1g / L, NaCl2.5g / L, pH 7.0-7.2.

[0034] Inoculation and culture conditions: Inoculate 5% of the seed liquid into the fermentation medium with a liquid volume of 10 mL, and culture at 37°C for 6 days.

[0035] The measured MK-7 content is 66.62mg / L, during this period, the metabolic law...

Embodiment 3

[0037] Fermentation medium: corn flour liquefaction 1L (reducing sugar content is about 60g / L), soybean meal powder 40g / L, soybean peptone 15g / L, yeast extract 15g / L, K 2 HPO 4 0.1g / L, NaCl2.5g / L, pH 7.0-7.2.

[0038] Inoculation and culture conditions: Inoculate 5% of the seed liquid into the fermentation medium with a liquid volume of 10 mL, and culture at 37°C for 5 days.

[0039] The measured MK-7 content was 62.26mg / L.

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Abstract

The invention relates to a strain of bacillus amyloliquefaciens capable of generating a great amount of MK-7 (menadione-7) in a culture process and a culture method thereof. The name of the strain is Y-2, and the class name is bacillus amyloliquefaciens. The obtained strain has stable hereditary character and stable yield trait, is suitable for industrial popularization, and is collected in the China Center for Type Culture Collection (CCTCC), with the collection number CCTCC No.M2013493. The invention also discloses a method for producing MK-7, which comprises the following steps: with 5% of seed inoculation amount, inoculating a seed solution into a fermentation medium; culturing for 2 hours at a rotation speed of 100r / min and a temperature of 37 DEG C; performing stationary culture for 6 days, wherein the highest yield of vitamin K2 (MK-7) reaches 66.62mg / L. According to the MK-7 high-yielding strain and production method provided by the invention, the production cycle is short, the yield is high, and feasibility is provided to the production of MK-7 through industrial fermentation.

Description

technical field [0001] The present invention relates to a strain of Bacillus amyloliquefaciens capable of producing a large amount of menadione-7 (MK-7) during the culture process, in particular to a Bacillus amyloliquefaciens capable of producing a large amount of MK-7 and its The production method belongs to the field of microbial fermentation. Background technique [0002] K vitamins are one of the four fat-soluble vitamins necessary for the human body, and can be divided into K according to different sources. 1 and K 2 Two kinds, of which K 1 Mainly derived from plants, also known as phylloquinone, K 2 Mainly derived from microorganisms, also known as menadione. Structurally, vitamin K 2 In addition to the naphthoquinone parent ring, there is also a side chain composed of isoprene units at the C-3 position. According to the difference in the number of isoprene units, vitamin K 2 Divided into 14 types, with MK -n Indicates that MK-7 (menadione-7) means vitamin K wi...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/66C12R1/07
Inventor 张伟国钱和严为留
Owner JIANGNAN UNIV
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