Bacillus natto and application of Bacillus natto in fermentation production of vitamin K2

A technology of Bacillus natto and vitamins, which is applied in the directions of fermentation, microorganism-based methods, bacteria, etc., can solve the problems of not mentioning the fermentation index of vitamin K2, the high price of vitamin K2, and the increase of production costs, and achieve the effect of strengthening oxygen supply. , The effect of reducing fermentation cost and low production cost

Active Publication Date: 2015-02-04
ZHEJIANG BAISHANZU BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The patents on Bacillus natto mainly focus on: (1) preparation of feed additives by solid-state fermentation; (2) preparation of functional products such as nattokinase and synbiotics; only two are related to this patent, and CN 102827798A discloses a Bacillus natto, which can produce high vitamin K2, mainly improves the growth rate of strains and VK2 production through ultraviolet, chemical mutagenesis and protoplast fusion methods; CN103898175A also discloses a Bacillus natto and a method for purifying vitamin K2. The main points of the invention are In the purification method, the fermentation index of vitamin K2 is not mentioned
The above patents did not study in detail the key factors that directly affect the accumulation of vitamin K2, resulting in low yield and difficult industrialization
Low production efficiency increases production costs, making vitamin K2 from microbial sources expensive

Method used

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  • Bacillus natto and application of Bacillus natto in fermentation production of vitamin K2
  • Bacillus natto and application of Bacillus natto in fermentation production of vitamin K2
  • Bacillus natto and application of Bacillus natto in fermentation production of vitamin K2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1: strain screening

[0043] Dissolve 1g of natto product in 100ml of normal saline, oscillate and mix evenly, and dilute the sample suspension 10 times in order to prepare 10 -1 -10 -7 concentration, respectively, 10 -3 -10 -7 Draw 100ul of the dilutions from the five concentrations and add them to the screening medium for coating, apply three plates for each concentration, incubate for 48 hours at 37°C, select larger colonies that can produce hydrolysis circles, and continue to streak in the screening medium , to obtain pure bacteria, after fermentation and re-screening, high-performance liquid phase analysis and mass spectrometry identification, the bacterial strain Bacillus natto R capable of producing vitamin K2 was obtained, and the vitamin K2 produced by the bacterial strain was in the MK-7 configuration (such as figure 1 ,2).

[0044] In order to improve the ability of the bacteria to decompose protein, the obtained Bacillus natto R was subjected ...

Embodiment 2

[0047] Example 2: Investigation on the basic growth performance of bacterial strains.

[0048] The bacterial strain CCTCC M 2014405 that screening obtains is carried out basic performance investigation, adopts different liquid seed culture medium to cultivate, investigates cell growth rate, and the result is shown in Table 1, and the culture medium used is (% represents the quality g of substance contained in every 100ml culture medium) :

[0049] Medium 1: 100mL bean sprout juice, 5% sucrose

[0050] Medium 2: 1% glucose, 1% peptone, 0.5% NaCl

[0051] Medium 3: 3% glucose, 4% peptone, 0.5% NaCl, 0.5% beef extract, 0.5% yeast extract

[0052] Medium 4: 0.5% glucose, 1% peptone, 0.5% NaCl, 0.5% beef extract, 0.5% yeast extract

[0053] Table 1 The growth of bacteria in different seed media

[0054]

[0055] From the above test results, except for the No. 2 medium, the other three can make the bacteria grow better, and the No. 3 medium is selected.

Embodiment 3

[0056] Example 3: Investigation on the ability of bacterial strains to accumulate vitamin K2.

[0057] Four different fermentation media were selected for fermentation, and the production capacity of vitamin K2 of the strain was preliminarily discussed. The culture temperature is 30° C., and the rotation speed is 120 rpm. (% represents the quality g of the substance contained in every 100ml medium):

[0058] Medium 1: 5% glycerol, 3% soybean peptone, 0.6g / L yeast powder, 0.3g / L K 2 HPO 4 , 0.1g / LCaCl 2 ·H 2 O, 0.3g / LMgSO 4 ·7H 2 o

[0059] Medium 2: 5% glycerol, 3% soybean powder, 0.6g / L yeast powder, 0.3mol / LK 2 HPO 4 , 0.1g / LCaCl 2 ·H2O, 0.3g / LMgSO 4 ·7H2O

[0060] Medium 3: 15g / L peptone, 25g / L yeast powder, 70g / L glycerol, 0.5g / L K 2 HPO 4 , pH7.3

[0061] Medium 4: 100mL soybean milk, 2% glucose, 0.5% NaCl, 2% NaOH to adjust pH7.0

[0062] Table 2 Comparison table of vitamin K2 produced by different fermentation media

[0063]

[0064] From the above t...

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Abstract

The invention discloses bacillus natto which is classified and named bacillus natto and has been preserved in the China Center for Type Culture Collection with the preservation number of CCTCC M 2014405. The strain is extracted from a natto product purchased in the market and is obtained through ultraviolet mutagenesis and screening. By the use of the strain, soybean protein can efficiently be utilized to synthesize vitamin K2. The invention also discloses a method for producing vitamin K2 grease by the utilization of the above bacillus natto. According to the invention, a strain fermentation technology is optimized through a novel oxygen supply strategy, and fermentation cylinder scale-up is carried out. The final output of vitamin K2 reaches 60.54 mg / L. Conversion rate relative to soybean protein is 2.018 mg / g. Production costs are low. By the above method, efficient production of vitamin K2 grease can be realized; the whole process is simple; operation is convenient; production cycle is short; content of vitamin K2 is high; and fermentation cost is reduced. The method is suitable for industrial production.

Description

technical field [0001] The invention relates to a bacillus natto and its application in fermenting and producing vitamin K2, belonging to the field of biotechnology. Background technique [0002] Vitamin K2 is a kind of fat-soluble blood coagulation vitamin, which is a series of menadione compounds. Because of its extremely small content in food, it is known as the "platinum vitamin" and has significant effects in the prevention and treatment of osteoporosis. Effect, known as "the fourth generation of anti-osteoporosis products." The biological activity of vitamin K2 is mainly reflected in promoting the production of prothrombin and increasing the synthesis of osteocalcin. It plays an important role in blood coagulation and bone metabolism. It can be used to treat and prevent hemorrhage, arterial calcification, bone Diseases such as arthritis and osteoporosis are superior to other vitamin K in increasing human bone density. In recent years, the research and understanding o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P7/66C12R1/07
CPCC12N1/20C12P7/66C12N1/205C12R2001/07
Inventor 黄和邱宏伟任路静王星丽
Owner ZHEJIANG BAISHANZU BIOTECH CO LTD
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