Bacillus amyloliquefaciens strain and microbial agent and preparation method of microbial agent

A technology of amyloliquefaciens and bacilli, which is applied in the field of microorganisms, can solve problems such as difficulty in achieving standardized control, difficult cost, and uneven tenderization effects, and achieve high food safety, simple preparation methods, and obvious effects

Inactive Publication Date: 2017-05-24
HUNAN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

The physical tenderization method mainly includes high-pressure tenderization method, which has the advantages of obvious effect and uniform effect, does not increase the chance of microbial contamination, and can also play a bactericidal effect; organic acids, calcium chloride, and phosphate Improving the tenderness of beef also has a certain impact; enzymatic tenderization is a hot spot in recent research, it is easy to use, and the effect is remarkable. The main enzyme of this method is protease, which is extracted from plants and cultured by microorganisms. The main enzymes extracted from plants are papain, Ficin and bromelain can degrade myofibrils, thereby improving the tenderness of meat. However, due to the limitation of raw materials, the practical application of this kind of protease has certain difficulties due to cost issues.
Proteases from microbial sources have different effects on meat tenderization, and it is difficult to achieve standardized control

Method used

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  • Bacillus amyloliquefaciens strain and microbial agent and preparation method of microbial agent
  • Bacillus amyloliquefaciens strain and microbial agent and preparation method of microbial agent

Examples

Experimental program
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Embodiment 1

[0027] A strain of Bacillus amyloliquefaciens, which the inventor isolated from Guizhou traditional fermented soya bean and sweet wine koji. After the study of the morphological characteristics of the strain, it was used in beef products, showing the advantages of the strain as a special tenderizer for meat, and has great potential to be developed into a commercial food tenderizer. There were 22 strains of protease-producing bacteria, and one strain with higher enzyme activity was screened out, named Bacillus amyloliquefaciens D7, with an enzyme activity of 71.68 U / g.

[0028] Bacillus amyloliquefaciens D7, on the isolation medium, the strain D7 is a white, smooth and moist colony, slightly raised, with folds in the middle of the colony, it is a spore-forming Gram-positive bacillus, figure 1 It is the Gram stain characteristic map under the electron microscope of the Bacillus amyloliquefaciens strain. Bacillus amyloliquefaciens D7 is film-like on the slope, with a white surfa...

Embodiment 2

[0032] A kind of Bacillus amyloliquefaciens bacterial agent, adopts following preparation method to prepare:

[0033] (1) Pick the Bacillus amyloliquefaciens strain from the slant culture medium in Example 1, inoculate it into the broth culture medium, and culture it on a shaker at 220 r / min for 12 hours at 37° C. to activate to obtain an activation solution.

[0034] (2) The activation solution was inoculated into the fermentation medium according to the inoculation amount of 3%, and cultured on a shaker at 220 r / min for 48 hours at 37° C. to obtain the Bacillus amyloliquefaciens bacterial agent.

[0035] Fermentation medium formula: glucose: 16.8g, yeast extract juice: 20g, NaCl: 2g, CaCl: 2g, dipotassium hydrogen phosphate: 5g, potassium dihydrogen phosphate: 1.25g, magnesium sulfate: 0.01g, iron sulfate: 0.001g , Distilled water: 1000mL, pH value: 7.66.

[0036] The Bacillus amyloliquefaciens inoculum of Example 2 is applied to beef tenderization. The specific steps are: ...

Embodiment 3

[0042] A preparation method of bacillus amyloliquefaciens bacterial agent, comprises the following steps:

[0043](1) Isolation of Bacillus amyloliquefaciens from Guizhou traditional tempeh and sweet wine koji. The strain is white, smooth and moist colony, slightly raised, with folds in the middle of the colony, which is a spore-forming Gram-positive bacillus, and a clearly visible transparent circle will be formed around it. It is filmy on slopes and has a white surface.

[0044] (2) The strain was purely cultured with an isolation medium, cultured in a biochemical medium box at 37°C for 24 hours, and then stored on a slant.

[0045] Separation medium formula: peptone: 10g, glucose: 1g, NaCl: 5g, CaCl: 0.1g, L-tyrosine: 0.1g, agar: 15g, casein: 5g, distilled water: 1000mL; pH value: 7.2~7.4 .

[0046] (3) Pick the strain of Bacillus amyloliquefaciens from the slant culture medium, inoculate it into the broth medium, culture it on a shaker at 220 r / min at 37°C for 12 hours,...

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Abstract

The invention provides a bacillus amyloliquefaciens strain, a microbial agent and a preparation method of the microbial agent. The bacillus amyloliquefaciens strain is Bacillus amyloliquefaciens D7 and collected in the General Microbiological Center of China Microbiological Culture Collection Management Committee on November 10th, 2016, and the collection number is CGMCC No.13250. The strain is activated and subjected to fermentation culture to prepare the microbial agent of the bacillus amyloliquefaciens strain, and the preparation method of the microbial agent comprises the following steps: inoculating the bacillus amyloliquefaciens strain into a broth culture medium to be activated so as to obtain an activation solution; and inoculating the activation solution to a fermentation culture medium to be subjected to the fermentation culture so as to obtain the microbial agent of the bacillus amyloliquefaciens. The bacillus amyloliquefaciens strain disclosed by the invention can produce proteases, and is obvious in tenderizing effect on beef products.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a strain of Bacillus amyloliquefaciens, a bacterial agent and a preparation method of the bacterial agent. Background technique [0002] With the change of people's meat structure and the continuous improvement of living standards, beef is more and more favored by consumers, and the tenderness of beef is the most important factor affecting consumers' evaluation of beef quality, so the relevant research on meat tenderization It has gradually become one of the hot spots that people pay attention to. In China, some cattle, buffaloes, and aged cattle raised in mountainous areas are also used as meat in due course. The texture of this type of beef is relatively poor, but it is an important supplement to the improvement of China's meat structure. Especially in underdeveloped areas and restaurants below mid-range, there is a large market, so it is of great practical significance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/07
CPCC12N1/20C12N1/205C12R2001/07
Inventor 李宗军侯爱香谭雅
Owner HUNAN AGRICULTURAL UNIV
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