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Method for detecting trypsin inhibitor activity

A trypsin inhibition and detection method technology, applied in the field of enzyme catalysis/enzyme inhibition biology, can solve the problems of poor reaction sensitivity and incomplete effect, and achieve the effects of convenient operation, thorough reaction and high reaction sensitivity

Inactive Publication Date: 2008-06-04
SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of the prior art is that the action site of trypsin in bovine serum albumin is an amide bond, the action is not complete, and the reaction sensitivity is poor

Method used

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  • Method for detecting trypsin inhibitor activity
  • Method for detecting trypsin inhibitor activity
  • Method for detecting trypsin inhibitor activity

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2

[0034] The difference from Example 1 is that

[0035] The detection method of trypsin inhibitor activity: after the reaction time of the trypsin reaction system is 12 minutes, the reaction temperature is 38° C., wherein the substrate of trypsin is BAPNA solution, and the amount added is 1.0 ml per milliliter of the trypsin reaction system. Described reaction termination liquid is acetic acid solution, and its addition is 0.2ml.

Embodiment 3

[0037] The difference from Example 1 is that

[0038] The detection method of trypsin inhibitor activity: after the reaction time of the trypsin reaction system is 15 minutes, the reaction temperature is 38° C., wherein the substrate of trypsin is BAPNA solution, and the amount added is 0.8ml per milliliter of the trypsin reaction system. Described reaction termination liquid is acetic acid solution, and its addition is 0.15ml.

[0039] Example 3

[0040] The difference from Example 1 is that

[0041] The detection method of trypsin inhibitor activity: after the reaction time of the trypsin reaction system is 14 minutes, the reaction temperature is 36°C, wherein the substrate of trypsin is BAPNA solution, and the amount added is 0.6ml per milliliter of the trypsin reaction system. Described reaction termination liquid is acetic acid solution, and its addition amount is 0.1ml.

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Abstract

The invention relates to the enzyme catalysis / enzyme inhibition biotechnological field, in particular to a test method for activity of a trypsin inhibitor. The particular test process is mainly that: a trypsin inhibitor extract and a trypsin action substrate are respectively added into a trypsin reaction system; after reaction for 10 to 15 minutes, reaction stopping solution is added, and then absorbance of the reaction stopping solution in the wavelength of 410 nautical miles is measured; a curve is drafted according to addition and the absorbance of the inhibitors; the activity of the trypsin inhibitor is determined; wherein, the trypsin action substrate is BAPNA solution and the addition is 0.5 to 1.0 milliliters per milliliter trypsin reaction system. The invention has the advantages of simple instruments used, convenient operation and high reaction sensitivity, and is suitable for testing activity of various trypsin inhibitors.

Description

technical field [0001] The invention relates to the field of enzyme catalysis / enzyme inhibition biotechnology, in particular to a method for detecting trypsin inhibitor activity. technical background [0002] The short-chain alkaloids contained in plant fruits (such as winter melon seeds, etc.) have trypsin inhibitor activity. It has been reported that these special alkaloid components extracted from wax gourd seeds can show weight loss effect under physiological conditions. In addition, winter melon seeds also have a variety of other medicinal functions, such as treating schistosomiasis with winter melon seed extract. Therefore, winter melon seeds and their extracts are widely accepted as a good health product. [0003] However, there is a lack of sensitive and simple methods for the determination of the trypsin inhibitor activity of wax gourd seed extracts. The existing determination method is to use bovine serum albumin as the substrate of trypsin, then add inhibitor c...

Claims

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Application Information

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IPC IPC(8): C12Q1/37
Inventor 徐慧李启平张忠泽张颖李慧史荣久韩斯琴杨伟超
Owner SHENYANG INST OF APPLIED ECOLOGY - CHINESE ACAD OF SCI
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