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Method for extracting protease A inhibitor and its application

An inhibitor, protease technology, applied in the direction of protease inhibitors, peptide preparation methods, chemical instruments and methods, etc., can solve the problems of practical application limitation, large addition amount, etc., and achieve high inhibitory effect, low addition amount, and extraction process. easy effect

Inactive Publication Date: 2004-11-17
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been some researches on the early strains and technology at home and abroad, but people have found that this problem cannot be completely solved, so looking for specific PrA inhibitors, as a post-modification idea, has begun to attract people's attention
[0006] Domestic research has attempted to extract PrA inhibitors from natural raw materials such as soybeans, peanuts, broad beans, soybeans, and barley. As a result, an inhibitor with a molecular weight of 5KD was obtained from soybeans, but its practical application was limited due to its large addition amount. limit

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Add water to the whole powder of Ganoderma lucidum at a ratio of 1:30, heat and extract at 55°C for 1 hour, centrifuge at 5000r / min for 10 minutes, and obtain the filtrate. The protein content of the whole powder of Ganoderma lucidum determined by Kjeldahl method is 10%. This extraction condition The extraction rate of protein is 85%, and filtrate carries out ethanol fractionation precipitation, and graded concentration section is 30%~80%, gets thick PrA inhibitor (lyophilized powder is 32% of whole powder weight, and wherein protein accounts for 12%, sugar accounted for 81%), according to the ratio of enzyme pH ratio of 23:1 added in the non-sterilized beer, at the same time with the same batch of beer without adding as a control, placed at room temperature (20 ℃), the result can make 15 days of beer brewing The rate of sustained decline slowed down by an average of 55%.

Embodiment 2

[0029] Example 2: Add water to the whole powder of Ganoderma lucidum at a ratio of 1:20, extract at 55°C for 1 hour, and centrifuge at 5000r / min for 10 minutes to obtain the filtrate. The protein content was determined by Kjeldahl method, and the extraction rate of protein under this extraction condition was found was 78%, the filtrate was subjected to ethanol fractionation precipitation, and the fractional concentration section was 30% to 65%, to obtain a crude PrA inhibitor (the freeze-dried powder is 30% of the whole powder weight, wherein the protein accounts for 12%, and the sugar accounts for 80%). Add it to unsterilized beer at a ratio of 25:1, and use the same batch of beer without adding it as a control, and place it at room temperature (20°C). Slowed down by 53.5%.

Embodiment 3

[0030] Example 3: Add water to the whole Ganoderma lucidum powder at 1:30, heat and extract at 55°C for 2 hours, centrifuge at 5000r / min for 10 minutes, and measure the protein content of the filtrate by Kjeldahl method. It is found that the extraction rate of protein under this extraction condition is 81%, the filtrate carries out ethanol graded precipitation, graded concentration section is 30%~80%, gets thick PrA inhibitor (lyophilized powder is 33% of whole powder weight, wherein protein accounts for 10%, sugar accounts for 85%), according to Add the enzyme pH ratio of 24:1 to non-sterilized beer, and take the same batch of beer without adding it as a control, and place it at room temperature (20°C). 55.8%.

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PUM

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Abstract

The invetion relates to a process for extracting proteinase A (PrA) from lucid ganoderma submerged cultuse liquid or lucid ganoderma baking powder and its use, the process comprises obtaining crude PrA suppression agent from the raffinate of lucid ganoderma fermentation liquid or baking powder through ethanol grading or film filtration grading, then obtaining refined PrA depressant by molecular screen chromatography to collect 30KD-40KD and partial cryodesiccation. The depressant can be applied to the production of pure draught beer.

Description

technical field [0001] An extraction method and application of a protease A inhibitor relate to the extraction and application of a protease A (PrA) inhibitor from ganoderma submerged fermentation liquid or ganoderma fermented whole powder. technical background [0002] The freshness and taste of pure draft beer are loved and pursued by people day by day, but real pure draft beer often has some enzymes left due to unpasteurization, and protease A (PrA) wherein will destroy beer foam protein Z, thereby make The foam retention of pure draft beer is reduced. [0003] Generally speaking, there are three main sources of protease in the beer brewing process. One is derived from malt. When barley germinates, it will produce natural protease, which can degrade protein and produce enough polypeptides with foam stability. But this enzyme is also removed after the wort is boiled, and there will be no residue. The second source is artificially added proteases, such as papain, which i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/14C07K14/81C12C5/02
Inventor 田亚平顾国贤章克昌李屹松周楠迪
Owner JIANGNAN UNIV
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