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Liquid culture medium for producing nattokinase in high yield

A nattokinase and culture medium technology, applied in the directions of enzymes, peptidases, hydrolase, etc., can solve the problems of short drug half-life, side effects, high price, etc., and achieve high unit enzyme activity, simple production process, and simple components. Effect

Pending Publication Date: 2017-12-15
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, with the improvement of living standards, more high-fat, high-sugar, and high-cholesterol foods are included in the dining table, and poor diets aggravate the occurrence of thrombotic diseases
At present, dissolving thrombus is an important means to treat this kind of diseases. The drugs used in clinical treatment today include streptokinase, urokinase, tissue-type plasminogen activator, single-chain urokinase-type plasminogen activator, Chemically modified plasmin-streptokinase activator complex, etc., but these drugs have obvious disadvantages: high price, short drug half-life, side effects of systemic bleeding, etc. Therefore, the development of new thrombolytic drugs has become an important research topic at present one
[0004] For many years, most of the food culture medium containing nattokinase has been produced by solid-state fermentation, and the solid-state fermentation process has the defects of easy infection of bacteria, difficult heat dissipation and difficult recovery, and cannot meet the production requirements of related medicines and health products; the current industrial production The main method of nattokinase is liquid fermentation, and the research on liquid fermentation is mostly focused on the breeding of excellent strains, the optimization of fermentation process and the separation and purification of products. There are few studies on the screening of liquid medium components, so screening A liquid medium with rich sources, simple components, low cost, high enzyme activity per unit of nattokinase, and simple production process is of great significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] The composition of the culture medium is: 4.0% of red kidney bean powder, 4.0% of glucose, 1.5% of sodium chloride, 0.1% of dipotassium hydrogen phosphate, 0.1% of potassium dihydrogen phosphate, and a pH value of 7.0, all w / v.

[0015] Carry out the liquid state fermentation of Bacillus subtilis (Bacillus subtilis subsp.UJS-1, CGMCC No.14434) with the medium prepared by the above-mentioned composition ratio, 37 ℃, 225rpm, fermentation 48h, by agarose-fibrin plate method (referring to Xie Pinghui etc., 2010, the fibrin plate method was used to measure nattokinase activity in health food) and the fermented liquid nattokinase enzyme activity was recorded as 33160u / ml.

Embodiment 2

[0017] The culture medium consists of: red kidney bean powder 5.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, all w / v.

[0018] Use the medium formulated with the above-mentioned ingredient ratios for liquid fermentation of Bacillus subtilis (Bacillus subtilis subsp. UJS-1, CGMCC No. 14434), 37 ° C, 225 rpm, 48 hours of fermentation, and the fermentation was measured by the agarose-fibrin plate method. The enzyme activity of liquid nattokinase is 29170u / ml.

Embodiment 3

[0020] The culture medium consists of: red kidney bean powder 6.0%, glucose 4.0%, sodium chloride 1.5%, dipotassium hydrogen phosphate 0.1%, potassium dihydrogen phosphate 0.1%, pH value 7.0, all w / v.

[0021] Use the medium formulated with the above-mentioned ingredient ratios for liquid fermentation of Bacillus subtilis (Bacillus subtilis subsp. UJS-1, CGMCC No. 14434), 37 ° C, 225 rpm, 48 hours of fermentation, and the fermentation was measured by the agarose-fibrin plate method. The enzyme activity of liquid nattokinase is 26720u / ml.

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PUM

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Abstract

The invention discloses a liquid culture medium for producing nattokinase in high yield and belongs to the technical field of biology. A fermenting culture medium comprises the following components: 4.0% of red kidney bean powder, 4% of glucose, 1.5% of sodium chloride, 0.05-0.2% of dipotassium phosphate and 0.05-0.2% of monopotassium phosphate. The PH value is 7.0-7.4. Compared with the prior art, the culture medium has the advantages of simple components, abundant sources and low cost; the unit enzyme activity of the fermented nattokinase is high; the manufacturing technology is simple; the enzyme activity of the fermented nattokinase reaches up to 3160u / ml.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a liquid culture medium with high yield of nattokinase. Background technique [0002] Thrombotic disease is a high incidence of middle-aged and elderly people, and has become one of the main causes of death and disability. As of 2016, my country's elderly population over the age of 60 was 220 million, accounting for 16.1% of the total population, and China has entered an aging society. In addition, with the improvement of living standards, more high-fat, high-sugar, and high-cholesterol foods are included in the dining table, and poor diets aggravate the occurrence of thrombotic diseases. At present, dissolving thrombus is an important means to treat this kind of diseases. The drugs used in clinical treatment today include streptokinase, urokinase, tissue-type plasminogen activator, single-chain urokinase-type plasminogen activator, Chemically modified plasmin-streptokinase activator...

Claims

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Application Information

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IPC IPC(8): C12N9/56C12R1/125
CPCC12N9/54C12Y304/21062
Inventor 龚爱华朱沛煌严永敏曾建彭琬昕杜凤移崔恒林
Owner JIANGSU UNIV
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