Industrial liquid fermentation method of alkaline protease

A technology of liquid fermentation and protease, applied in the field of bioengineering, can solve the problems of high production costs, meet the needs of the market, the process is easy to operate, and the effect of optimizing the fermentation mechanism

Pending Publication Date: 2019-09-17
江苏邦臣生物科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the enzyme activity of alkaline protease that can be industrially produced in the market is generally low, making the production cost corresponding to the unit enzyme activity relatively high, which is far from meeting the needs of the current detergent and other industries

Method used

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  • Industrial liquid fermentation method of alkaline protease
  • Industrial liquid fermentation method of alkaline protease

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Experimental program
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Effect test

Embodiment 1

[0021] The bacterial strain of alkaline protease of the present embodiment is the industrialized liquid fermentation method of bacterial classification, comprises the steps:

[0022] ① No. seed cultivation: Insert the bacterial strains capable of producing alkaline protease into a 1500 ml shake flask, choose Bacillus alkalophilus here, the medium loading capacity is 100 ml, the rotary shaker is 220 rpm, and the culture temperature is 36- 38°C, culture time 6-15h;

[0023] Cultivation of No. ② seeds: Inoculate No. ① seeds into 500 ml shake flasks according to 10% inoculum amount, the medium capacity is 100 ml, the rotary shaker is 220 rpm, the culture temperature is 36-38 °C, and the culture time is 6-6. 15h; culture conditions are the same as the first-class seeds;

[0024] Then carry out fermentation culture.

[0025] The medium used for fermentation is: fermentation medium g / L: yeast extract powder 20, cottonseed cake powder 30, maltodextrin 100, sodium citrate 3, calcium ...

Embodiment 2

[0034] The bacterial strain of alkaline protease of the present embodiment is the industrialized liquid fermentation method of bacterial classification, comprises the steps:

[0035] ① Seed cultivation: Inoculate the strain of Bacillus alkalophilus into a 500ml shaker flask, with a medium capacity of 100ml, a rotary shaker at 240 rpm, a culture temperature of 36°C, and a culture time of 15 hours;

[0036] Cultivation of No. ② seed: Inoculate No. ① seed into a 500 ml shake flask according to 10% inoculum amount, the medium capacity is 100 ml, the rotary shaker is 240 rpm, the culture temperature is 36°C, and the culture time is 15 hours; culture conditions Same as Tier 1 seed;

[0037] Cultivation of No. ③ seed: Inoculate No. ② seed with 10% inoculum amount into a 500 ml shake flask, the medium capacity is 1000 ml, the rotary shaker is 100 rpm, the culture temperature is 34° C., and the culture time is 10 h.

[0038] Then carry out fermentation culture.

[0039] The medium us...

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Abstract

The invention relates to the technical field of bioengineering, in particular to an industrial liquid fermentation method of alkaline protease. The method comprises the following steps of culture of first seeds, wherein a strain is inoculated into a 1500 ml shake flask, and 100 ml of culture medium is contained; culture of second seeds, wherein the first seeds are inoculated into a 500 ml shake flask according to the inoculation amount of 10%, 100 ml of culture medium is contained, and culture conditions of the second seeds and culture conditions of the first seeds are the same; fermentation culture is carried out. A corresponding fermentation mechanism is optimized, the fermentation mechanism is simple and convenient to operate, the culture conditions are mild, the activity of fermentation enzyme is 100000 u / ml or above, the production cost of unit enzyme activity is reduced, and requirements of the market are greatly met. The alkaline protease can be widely applied to the industries of detergents, feed, leather manufacturing and the like.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to an industrialized liquid fermentation method of alkaline protease. Background technique [0002] Alkaline Protease is a protease with an optimum pH of 9-11, with a molecular weight of 26,000-34,000 Daltons, and is generally stable at a pH of 5-10. Because its active center contains serine (Ser), it belongs to serine protease. It not only hydrolyzes peptide bonds, but also has the functions of hydrolyzing ester bonds, amide bonds, transesterification and transpeptide. Widely used in detergent, medical, silk, leather and other industries. [0003] In recent years, with the improvement of people's living standards and the needs of environmental protection, phosphorus-containing detergents have been basically eliminated, replaced by a large number of applications of various enzyme-added detergents. Because the aqueous solution of detergent is generally alkaline, with a pH o...

Claims

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Application Information

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IPC IPC(8): C12N9/54C12R1/07
CPCC12N9/54
Inventor 陈永忠陈圣卿
Owner 江苏邦臣生物科技股份有限公司
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