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Alkaline protease

A protease and alkaline technology, applied in the field of alkaline protease, can solve the problems of low hydrolysis rate and achieve high hydrolysis efficiency, high degree of hydrolysis, and strong adaptability

Inactive Publication Date: 2009-07-22
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The enzymatic method has strong selectivity, but the hydrolysis rate is low

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Fermentation of hair enzyme and extraction of crude enzyme

[0028] 1.1 Strains

[0029] Mucor actinosa AS3.2778 was purchased from the General Microbiology Center of China Committee for the Collection of Microorganisms.

[0030] 1.2 Seed medium and seed activation

[0031] Add 10g of bran and 10ml of water into a 250ml triangular flask, stir and mix evenly with a glass rod, and smash larger agglomerates (try to make the particles uniform). Sterilize at 121°C for 30 minutes, take it out from the heat, shake it up, and cool it down for later use.

[0032] The sterilized seed medium was inoculated with a ring of slant spores in a test tube per bottle, mixed evenly, and the surface of the medium was made as smooth as possible, and the thickness of the medium layer was consistent, and then placed in a biochemical incubator at 28°C for 3 days.

[0033] 1.3 Preparation of spore suspension

[0034] Under sterile conditions, add 100 ml of sterile water to the a...

Embodiment 2

[0043] The separation and purification of embodiment 2 alkaline protease

[0044] 2.1 Sectional salting-out of ammonium sulfate

[0045] Take a certain volume of crude enzyme solution, slowly add solid ammonium sulfate to 40% saturation on an ice-water bath, fully dissolve and let stand in a refrigerator at 4°C for 3 hours, centrifuge at 12000g / min for 20min at 4°C, take the supernatant and continue Add solid ammonium sulfate to 85% saturation, let it stand overnight in a refrigerator at 4°C, then centrifuge at 12000g / min for 20min at 4°C, dissolve the protein precipitate with 0.02M, pH7.5 Tris-HCl buffer, and dialyze for desalting.

[0046]2.2 DEAE-Sepharose anion exchange

[0047] Equilibrate the DEAE-Sepharose anion-exchange column with 0.02M, pH7.5 Tris-HCl buffer solution (the basis for good balance: the effluent pH7.5, the conductivity is stable), take a certain amount of enzyme solution after salting out and desalting, and add the sample anion exchange column. With 0...

Embodiment 3

[0066] Example 3 Determination of Catalytic Properties of Mucor Alkaline Protease Pb1

[0067] 3.1 Optimal action temperature of alkaline protease

[0068] According to the enzyme activity assay method, the activity of Mucor alkaline protease was measured at different temperatures (range of 30-70° C.), and the effect of temperature on the activity of Mucor alkaline protease was investigated. According to the experimental results, the temperature-activity curve was drawn, and the optimum action temperature of mucoralline protease was determined accordingly.

[0069] 3.2 The optimal pH of mucor alkaline protease

[0070] According to the enzyme activity assay method, under different pH buffer conditions (pH 3.0-5.0, 0.05M acetic acid buffered saline; pH6.0-7.0, 0.05M phosphate buffered saline; pH 8.0-9.0, 0.02M Tris-HCl; pH 9.5- 10.5, 0.02M glycine-NaOH) to measure the activity of Mucor alkaline protease, and investigate the effect of pH on the activity of Mucor alkaline prote...

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Abstract

The invention relates to an alkali protease, which can hydrolyze protein under alkali condition and has specified physicochemical characteristics and high hydrolysis percent.

Description

technical field [0001] The invention relates to an alkaline protease, in particular to an alkaline protease with high hydrolysis efficiency. Background technique [0002] Soybean contains 40% protein, which can provide 8 kinds of essential amino acids, vitamins and minerals needed by animals. Soybean protein resources are very rich, the cost of raw materials is low, and it has functional properties such as emulsification and foaming. But at the same time, soybean protein contains some antigenic substances that cause allergic reactions—allergenic factors. The main antigenic components of soybean protein that cause allergic reactions are glycinin and β-conglycinin. The main solution is to enzymatically hydrolyze soybean protein. After hydrolysis, the hydrolyzate of soybean protein is easier to digest and absorb than amino acids and proteins, and can also change its solubility and reduce viscosity. [0003] There are three methods of hydrolyzing soybean protein, acid method, ...

Claims

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Application Information

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IPC IPC(8): C12N9/58C12R1/785
Inventor 罗晓春潘进权王菊芳谢明权王小宁
Owner SOUTH CHINA UNIV OF TECH
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