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A strain of Bacillus licheniformis with high protease production and its fermentative enzyme production method

A technology of Bacillus licheniformis and protease, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of underutilized feather waste, waste liquid and gas polluting the environment, and difficult to be degraded. The effect of low raw material cost, high process yield and low total production cost

Active Publication Date: 2021-06-08
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, keratin cannot be degraded in the digestive tract of animals because it contains disulfide bonds, and it is difficult to degrade in vitro. The traditional physical and chemical treatment technology consumes a lot of energy, and the waste liquid and gas generated by the processing process pollute the environment.
The annual 2 million tons of feather waste in my country is not fully utilized, resulting in a waste of resources

Method used

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  • A strain of Bacillus licheniformis with high protease production and its fermentative enzyme production method
  • A strain of Bacillus licheniformis with high protease production and its fermentative enzyme production method
  • A strain of Bacillus licheniformis with high protease production and its fermentative enzyme production method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1, the acquisition, identification and preservation of Bacillus licheniformis GZ73

[0045] Wild bacteria were isolated from the high-temperature fermentation product of feather waste, and then subjected to repeated ultraviolet mutagenesis and nitrosoguanidine mutagenesis, and then a strain was obtained by screening, which was named strain GZ73.

[0046] Morphological characteristics of the strain GZ73: white colonies formed on LB solid medium, round and translucent, smooth in texture, with jagged edges; microscopically, the strain was thin rod-shaped, 0.5-2.0 μm wide, 1.0-9.0 μm long μm.

[0047] Physiological and biochemical characteristics of strain GZ73: positive for catalase, positive for growth NaCl 2%-7%, negative for phenylalanine deamination, positive for most of citrate utilization, negative for lecithinase, positive for casein hydrolysis, positive for starch hydrolysis, Methyl red test negative, glucose acid production positive, glucose gas productio...

Embodiment 2

[0053] Embodiment 2, the stability of bacillus licheniformis GZ73

[0054] Bacillus licheniformis GZ73 was subcultured on solid LB medium plate for several times, the strain obtained from the first passage was named as the F1 generation strain, the strain obtained from the second passage was named as the F2 generation strain, and the strain obtained from the third passage was named as the F2 generation strain. The bacterial strain is named F3 generation strain, the bacterial strain obtained by the fourth passage is named as the F4 generation strain, the fifth passage is named as the F5 generation strain, and the sixth passage is named as the F6 generation strain.

[0055] The tested strains are: F1 generation strain, F2 generation strain, F3 generation strain, F4 generation strain, F5 generation strain or F6 generation strain.

[0056] The test strain was inoculated into the seed medium (the seed medium was the same as in Example 3), and cultured with shaking at 34° C. and 200...

Embodiment 3

[0060] Embodiment 3, the ability of Bacillus licheniformis GZ73 to produce protease

[0061] 1. Preparation of culture medium

[0062] Solid medium: yeast powder 4g / L, peptone 5g / L, glucose 10g / L, beef extract 5g / L, potassium dihydrogen phosphate 10g / L, agar 15g / L, the balance is water; pH7.0-8.0; Sterilize at 121°C for 30 minutes.

[0063] Seed medium: yeast powder 8g / L, maltodextrin 120g / L, peptone 4g / L, corn steep liquor 50g / L, potassium dihydrogen phosphate 11g / L, magnesium sulfate 0.8g / L, the balance is water; pH7. 0-8.0; sterilize at 121°C for 30 minutes.

[0064] Fermentation medium: yeast powder 9g / L, corn flour 120g / L, soybean meal powder 25g / L, corn steep liquor 10g / L, calcium chloride 4g / L, potassium dihydrogen phosphate 12g / L, magnesium sulfate 0.8g / L, Sodium citrate 4g / L, medium temperature amylase, high temperature amylase, the balance is water; pH7.0-8.0. The preparation method of the fermentation medium: put water, corn flour, and soybean meal into the batc...

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Abstract

The invention discloses a strain of bacillus licheniformis with high protease production and a method for fermenting and producing the enzyme. The Bacillus licheniformis GZ73 provided by the present invention is referred to as Bacillus licheniformis GZ73 for short, and the preservation number is CGMCC No.18048. The invention also protects the application of Bacillus licheniformis GZ73 in the production of protease. The protease is prepared by fermenting Bacillus licheniformis GZ73, which has high enzyme production efficiency, low raw material cost, high enzyme activity of fermented liquid, high yield of enzyme extraction and purification process, and low total production cost. The enzyme preparation provided by the invention can be widely used in industries such as feed, food, detergent and the like.

Description

technical field [0001] The invention belongs to the field of bioengineering of enzyme preparations, and relates to a strain of bacillus licheniformis with high protease production and a fermentative enzyme production method thereof. Background technique [0002] Soybean meal is the most important feed protein raw material, and the annual consumption in my country is about 70 million tons. But soybean meal contains a variety of anti-nutritional proteins. According to the analysis and determination of the content of anti-nutritional factors (ANFs) (including glycinin, β-conglycinin, and trypsin inhibitor) in 342 batches of commercially available soybean meal by the Feed Institute of the Chinese Academy of Agricultural Sciences and the Feed Industry Center of China Agricultural University The results show that the content of ANFs in soybean meal is 50%-300% higher than that of extruded soybean, which seriously affects the normal digestion, absorption and utilization of soybean...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/56A23K20/189C12R1/10
CPCA23K20/189C12N9/54C12N1/205C12R2001/10
Inventor 丁宏标张荣庆陈守文乔宇郑春田
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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