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A strain of bacillus subtilis for producing lab ferment

A technology of Bacillus subtilis and chymosin, applied in the direction of enzymes, bacteria, and microorganism-based methods, can solve the problems of difficult development, strong proteolysis ability, and long growth cycle, and achieve good curdling effect

Inactive Publication Date: 2009-05-20
李红玉 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rennet is traditionally derived from rennet enzyme extracted from calf abomasum (4th stomach) [5] , but with the continuous development of the world cheese industry, the method of slaughtering a large number of calves every year to obtain rennet is obviously extremely inconsistent with the development of modern industry; vegetable rennet can be obtained from papaya, pineapple, fig, red Extract from flowers and other plants [6] , but its proteolytic ability is strong, and it is difficult to develop due to time, region, long growth cycle and other conditions; more than 40 kinds of microorganisms have been found to produce chymosin with a certain activity, and these microorganisms are mainly actinomycetes, bacteria and Fungus etc. [7]

Method used

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  • A strain of bacillus subtilis for producing lab ferment

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Example 1 Isolation of Bacillus subtilis QL-2 from cheese by dilution plate method:

[0062] (1) Preparation of cheese dilution: Weigh 5g of cheese, quickly pour it into 45ml of sterile water with glass beads, shake it on a shaker for 20min, let it stand for clarification, and it becomes 10 -1 suspension. Aseptically draw 0.5ml of its supernatant and add 4.5ml of sterile water to get 10 -2 Diluent. Repeat this and dilute to 10 with sterile water -3 、10 -4 、10 -5 、10 -6 、10 -7 A total of 7 concentrations.

[0063] (2) Cultivation: Add sterilized casein hydrolysis plates cooled to 45-50°C for each dilution (take 5g of skim milk powder and add it to 50mL of distilled water or use 50mL of skim milk, otherwise known as 1.5g of agar, dissolve it in 50mL of distilled water. The two liquids are sterilized separately. When it is cooled to 45-50°C, mix the two liquids and pour them into a flat plate to serve), and mix them immediately to make a flat plate. Incubate at 30...

Embodiment 2

[0067] Example 2 Characterization of Bacillus subtilis QL-2

[0068] (1). Bacterial morphology observation and staining reaction

[0069] Bacillus subtilis QL-2 was inoculated in NA solid medium and cultured at a constant temperature for a suitable time, then stained for microscopic examination. Including simple staining, Gram staining (crystal shoxenamide staining method), flagella staining (silver nitrate staining method), capsule staining (wet ink method), spore staining (modified Schaeffer-Fulton's staining method).

[0070] Staining results: Bacillus subtilis QL-2 is a rod-shaped bacterium, G + , flagella are perinatal, each cell has 8 to 16 flagella, and its length is about 1 to 2 times that of the cell; there is a capsule; a few spores are produced, mesozoic, and the sporangia are enlarged.

[0071] (2). Effects of environmental factors on Bacillus subtilis QL-2

[0072] o 2 Effect on Bacillus subtilis QL-2

[0073] Determination of O by the deep agar method 2 For...

Embodiment 3

[0125] Embodiment 3 Bacillus subtilis QL-2 produces the activity of chymosin and the curdling activity of its fermented liquid

[0126] (1).Activity of Bacillus subtilis QL-2 to produce chymosin

[0127] The culture of Bacillus subtilis QL-2 shaken at 30°C and 120rpm for 24h was inoculated into NA liquid medium (30ml / 250ml Erlenmeyer flask, the inoculum size was 1:30), and shaken at 30°C and 120rpm on a shaker. After culturing for 24 hours, the supernatant was centrifuged to measure the chymosin activity, and the chymosin activity of the fermentation liquid reached the highest 44.4SU.

[0128] 2. Content of rennet in Bacillus subtilis QL-2 fermentation broth

[0129] Using Maxiren chymosin as a standard, make a standard curve of rennet concentration and rennet activity. According to the chymosin activity of the fermentation broth when Bacillus subtilis QL-2 was cultured for 30 hours, the content of chymosin was calculated to be 5100 mg / L.

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Abstract

The invention relates to a bacillus subtilis for producing renninum. The bacillus subtilis strain is separated from cheese and preserved in China Center for Type Culture Collection (CCTCC) with the preservation number of M208023 and the name of bacillus subtilis QL-2 (Bacillus subtilis QL-2). A zymotic fluid of the bacillus subtilis QL-2 is rich in renninum, and has great renneting effect as well as the potential of being applied into renninum industrial production.

Description

technical field [0001] The present invention relates to a rennet-producing Bacillus subtilis QL-2 (Bacillus subtilis QL-2), which is isolated from cheese and has been preserved in the China Center for Type Culture Collection (CCTCC) with the preservation number M208023. The fermented liquid is rich in rennet and has good curdling effect, and has the potential to be used in the industrial production of rennet. Background technique [0002] Cheese is rich in a variety of nutrients, easy to digest and absorb, and not easy to cause fat, so it is regarded as an ideal food by nutritionists, known as "milk gold", [1] . Rennet is a key enzyme preparation for coagulating milk in cheese production [2] , which is an acid protease. The curdling process can be divided into two steps: the first step is to specifically hydrolyze the peptide bond between 105-106 phenylalanine and methionine in the κ-casein polypeptide chain in milk to form a stable paraκ-casein and hydrophilic glycomacr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/56C12R1/125
Inventor 李红玉刘河涛李洋
Owner 李红玉
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