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Bacillus licheniformis strain and use thereof

A technology of Bacillus licheniformis and strains, applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., to achieve the effects of high conversion rate, less by-products, and mild reaction conditions

Active Publication Date: 2011-10-19
KWEICHOW MOUTAI COMPANY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few reports on the production of 2,5-dimethylpyrazine by biocatalysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Bacillus licheniformis of the present invention ( Bacillus Licheniformis ) Separation steps of MTDB-02:

[0029] (1) Add 10 g of Moutai high-temperature Daqu koji powder into 90 g of sterile water (or sterile saline) filled with glass beads, shake on a shaker at 37°C and 200 rpm for 2 hours; apply gradient dilution to the plate, Cultivate at 37°C for 48 hours; select colonies with better growth and different characters from the plate and inoculate them on the slant of broth agar respectively, as strains for screening;

[0030] (2) Inoculate the strains obtained above for screening into the nutrient broth medium, culture at 37°C for 24 hours, collect the bacteria by centrifugation, add 50 g / L L-threonine and 90 mL transformation substrate solution, 37°C, 200 After 24 hours of rpm conversion, the conversion solution was analyzed by GC-MS for the concentration of 2,5-dimethylpyrazine, and strains with high 2,5-dimethylpyrazine production were selected to obtain MTDB-02. ...

Embodiment 2

[0038] A method for transforming and preparing 2,5-dimethylpyrazine with bacillus licheniformis MTDB-02, comprising the following steps:

[0039] (1) Expand the culture of Bacillus licheniformis grown on the slant of the test tube in a liquid shaker flask; then transfer 1% of the seeding amount into a 250 mL Erlenmeyer flask containing 20 mL of fermentation medium, and the shaker speed 100 rpm, culture time 24 hours, culture temperature 40 ℃, obtain fermentation liquid as the biocatalyst of L-threonine conversion reaction; Wherein the composition of fermentation medium is: glucose 0.1 g / L, peptone 20 g / L, yeast Paste 10 g / L, sodium chloride 0.1 g / L, potassium dihydrogen phosphate 0.1 g / L; the initial pH of the culture medium was 5.

[0040] (2) According to the L-threonine concentration of 1 g / L, the initial conversion pH is 5, and distilled water is used to prepare the substrate solution for biotransformation;

[0041] (3) Use a biocatalyst to transform the substrate solutio...

Embodiment 3

[0043] A method for transforming and preparing 2,5-dimethylpyrazine with bacillus licheniformis MTDB-02, comprising the following steps:

[0044] (1) Expand the culture of Bacillus licheniformis grown on the slant of the test tube in a liquid shaker flask; then transfer 10% of the seeding amount into a 250 mL Erlenmeyer flask containing 90 mL of fermentation medium, and the shaker speed 250 rpm, culture temperature 20°C, culture time 24 h, the fermentation broth was centrifuged and filtered to obtain wet bacteria as a biocatalyst for L-threonine conversion reaction; wherein the composition of the fermentation medium was: glucose 5 g / L , peptone 0.1 g / L, yeast extract 10 g / L, sodium chloride 10 g / L, potassium dihydrogen phosphate 10 g / L; the initial pH of the medium was 8.

[0045] (2) According to the L-threonine concentration of 50 g / L, the initial conversion pH is 8, and the biotransformation substrate solution is prepared with distilled water;

[0046] (3) Use a biocatalys...

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PUM

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Abstract

The invention discloses a bacillus licheniformis strain MTDB-02 and use thereof. The preservation number of the strain is CGMCC NO.4825. The characteristics at bacteriology are as follows: at the initial stage, the colony form is protruded and circular, and the surface of the colonial form is smooth and has luster, at the later period of the culture, the colony edge is irregular, the surface of the colony form is flat and light yellow, and has no luster, and the 16SsDNA gene sequence has 99% homology with the 16SrDNA gene sequence of multi bacillus licheniformis strains. The strain is used for converting L-threonine to prepare 2,5-dimethyl pyrazine, the reaction condition is mild, the harmful substance content is low, the influence of plantation is free, the large scale production can be carried out, and the production cycle is short.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a strain of Bacillus licheniformis and its use in transforming L-threonine to prepare 2,5-dimethylpyrazine. Background technique [0002] 2,5-Dimethylpyrazine is a common heterocyclic nitrogen-containing compound that widely exists in food raw materials, processed foods and alcoholic beverages. Aroma, with the smell of burnt protein and cocoa fruit, often used in the deployment of food flavors such as baked goods, cold drinks, meat, peanuts, potatoes, nuts, coffee, cocoa, etc., and the content is relatively high in Chinese sauce-flavored liquors . Pyrazines are considered to be characteristic substances in Maotai-flavor liquors, and they usually have a lower flavor threshold. [0003] L-threonine is an essential amino acid, and L-threonine is mainly used in medicine, chemical reagents, food fortifiers, feed additives, etc. It can be used as a precursor for biotransformation into...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P17/12C12R1/10
Inventor 杨帆王和玉林琳雷良波王莉杨代永吕云怀
Owner KWEICHOW MOUTAI COMPANY
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