Bacillus amyloliquefaciens D4 and method for producing chymosin by fermentation of same

A technology of amylolytic spores and chymosin, which is applied in the field of bioengineering and can solve the problems of difficult extraction of fermentation products, long fermentation period, and easy damage of mycelium

Inactive Publication Date: 2011-05-04
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, fungi such as Mucor oryzae and Aspergillus niger are often used in industrial production to ferment chymosin, which has the disadvantages

Method used

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  • Bacillus amyloliquefaciens D4 and method for producing chymosin by fermentation of same
  • Bacillus amyloliquefaciens D4 and method for producing chymosin by fermentation of same
  • Bacillus amyloliquefaciens D4 and method for producing chymosin by fermentation of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Scrape the two-ring strains from the inclined surface, inoculate them in a 250ml Erlenmeyer flask with 70ml seed medium for seed culture. The conditions are as follows: the culture time is 13 hours, the temperature is 37° C., and the rotation speed of the shaker is 240 r / min. Then insert 3% inoculum into a 250ml Erlenmeyer flask with 50ml fermentation medium for fermentation. The conditions are as follows: the fermentation time is 48 hours, the temperature is 37° C., and the rotating speed of the shaker is 180 r / min, and the fermentation liquid with an enzyme activity of 2310.2 SU / ml can be obtained.

[0078] The composition of the seed medium is: beef extract 3g / L, peptone 10g / L, NaCl 5g / L, pH 7.0;

[0079] The liquid submerged fermentation medium is composed by mass percentage: 18% of bran extract, 3% of glucose, 2% of skim milk, 0.2% of disodium hydrogen phosphate, 0.1% of casein, 0.10% of magnesium sulfate, and distilled water as the balance;

[0080] The preparat...

Embodiment 2

[0082] Scrape the two-ring strains from the inclined surface, inoculate them in a 250ml Erlenmeyer flask with 70ml seed medium for seed culture. The conditions are: the culture time is 13 hours, the temperature is 37° C., and the rotation speed of the shaker is 220 r / min. Then insert 3% inoculum into a 250ml Erlenmeyer flask with 50ml fermentation medium for fermentation. The conditions are as follows: the fermentation time is 48 hours, the temperature is 37° C., and the rotating speed of the shaker is 175 r / min, and the fermentation liquid with an enzyme activity of 2308.5 SU / ml can be obtained.

[0083] The composition of the seed medium is: beef extract 3g / L, peptone 11g / L, sodium chloride 5g / L, pH7.0;

[0084] The liquid submerged fermentation medium is composed by mass percent: 14% of bran extract, 5% of glucose, 2% of skim milk, 0.2% of disodium hydrogen phosphate, 0.1% of casein, 0.10% of magnesium sulfate, and distilled water as the balance. The preparation method of...

Embodiment 3

[0086] Scrape the two-ring strains from the inclined surface, inoculate them in a 250ml Erlenmeyer flask with 70ml seed medium for seed culture. The conditions are as follows: the culture time is 13 hours, the temperature is 37° C., and the rotation speed of the shaker is 200 r / min. Then insert 3% inoculum into a 250ml Erlenmeyer flask with 50ml fermentation medium for fermentation. The conditions are as follows: the fermentation time is 48 hours, the temperature is 37° C., and the rotating speed of the shaker is 170 r / min, and the fermentation broth with an enzyme activity of 2214.7 SU / ml can be obtained.

[0087] The composition of the seed medium is: beef extract 3g / L, peptone 12g / L, sodium chloride 5g / L, pH7.0;

[0088] Liquid submerged fermentation medium composition: 10% of bran extract, 8% of glucose, 2% of skim milk, 0.2% of disodium hydrogen phosphate, 0.1% of casein, 0.10% of magnesium sulfate, and distilled water as the balance. The preparation method of the bran ...

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Abstract

The invention discloses bacillus amyloliquefaciens D4 and a method for producing chymosin by fermentation of the same. The strain is screened from yak pasturing environmental soil of Gannan Tibetan Autonomous Prefecture of Gansu Province by adopting a casein culture medium, the chymosin can be obtained by deep fermentation of shake flask liquid, and the activity of the chymosin can reach 2,310.2SU/ml. The chymosin can be used for producing cheese and casein.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a strain of bacillus amyloliquefaciens D4 and a method for preparing rennet by using it. Background technique [0002] Rennet is a key enzyme in the cheese making process, its main function is to specifically cut the peptide bond of casein Phe 105-Met 106 to form stable para-κ-casein and hydrophilic glycogen peptide; when total κ-casein is about 85% hydrolyzed, at Ca 2+ Precipitation of casein micelles occurs in the presence of coagulated or coagulated milk formed by chemical bonds formed between the casein micelles. In order to meet the requirements of the rapid development of cheese and casein production in my country and to solve the supply contradiction of rennet, the research and development of rennet has become an urgent task for the dairy industry. The Ministry of Science and Technology of the National Development and Reform Commission listed the development of chym...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N9/54C12R1/07
Inventor 甘伯中张卫兵宋曦高维东贺晓玲
Owner GANSU AGRI UNIV
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