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Method and kit for detecting activity of nattokinase, and application of method

A nattokinase and detection method technology, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of high measurement cost, limited scope of application, harsh equipment and reagents, etc., and achieve high stability, The effect of high accuracy and small data fluctuation

Active Publication Date: 2018-04-13
GUANGZHOU BAIWO BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During the fermentation process of nattokinase, a large amount of subtilases will be produced, which will interfere with the determination of nattokinase activity
In addition, tetrapeptide substrates are expensive, making the assay cost prohibitive
[0008] However, the equipment and reagents required by the enzyme-linked immunosorbent assay (ELIZA) method are more demanding, and the scope of application is limited.

Method used

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  • Method and kit for detecting activity of nattokinase, and application of method
  • Method and kit for detecting activity of nattokinase, and application of method
  • Method and kit for detecting activity of nattokinase, and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Measurement of the activity value of nattokinase samples 1-3 to be tested

Embodiment 1

[0084] Use method of the present invention to measure the activity value of nattokinase sample 1~3 to be tested:

[0085] (1) Preparation of thrombus tube

[0086] Weigh 9mg of fibrinogen, dissolve it in 1mL of normal saline to form a fibrinogen solution, keep it warm in a water bath at 37°C for 5 minutes, add half a drop of red ink, shake well, dip the capillary for about 1mm of thrombin, and immediately continue to absorb the fibrinogen. The original protein solution is about 6cm, and then dip the non-absorbing end of the capillary with some molten wax to seal it. Stand still until the solution in the capillary solidifies to form a thrombus to obtain a thrombus tube, mark the height of the thrombus on the capillary, and record the length of the thrombus in the thrombus tube as the first thrombus length.

[0087] (2) Thrombolysis

[0088] Weigh the urokinase standard and dissolve it in normal saline to prepare 0U / ml, 10U / ml, 20U / ml, 30U / ml, 40U / ml, 50U / ml, 60U / ml, 70U / ml, 8...

Embodiment 2

[0098] The activity value of nattokinase sample 2 to be tested is determined in the same manner as in Example 1.

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Abstract

The invention relates to a method for detecting the activity of nattokinase. The method comprises the following steps: preparing a thrombus tube, dissolving a thrombus, determining the thrombolytic length of a standard solution, drawing a standard curve, and determining the nattokinase activity of a sample to be tested. The method has the advantages of high accuracy, low data fluctuation, high stability, small amount of the sample used in the method, no complicated instruments, and realization of simultaneous determination of multiple samples, and is suitable for high-throughput screening of nattokinase activators or inhibitors.

Description

technical field [0001] The invention relates to the technical field of kinase detection, in particular to a detection method, kit and application of nattokinase activity. Background technique [0002] Nattokinase is a kind of Bacillus subtilis protein kinase, which is an alkaline serine protease produced by Bacillus natto or Bacillus subtilis. times. [0003] Nattokinase has the functions of dissolving thrombus, reducing blood viscosity, improving blood circulation, softening and increasing blood vessel elasticity. In addition, the price per hectogram of natto is equivalent to urokinase at US$1,500. In addition, the half-life of urokinase is 4-20 minutes, while the activity of nattokinase can last for 8-12 hours, and nattokinase can inhibit the effect of fibrin Better than urokinase and plasmin, and relatively heat-resistant. Therefore, it can be seen that compared with other thrombolytic drugs, nattokinase has low cost, good safety, small molecular weight, good oral effe...

Claims

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Application Information

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IPC IPC(8): C12Q1/56C12Q1/37
CPCC12Q1/37C12Q1/56
Inventor 张子曦
Owner GUANGZHOU BAIWO BIOTECH CO LTD
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