Liquid fermentation method for producing natto kinase in high yield

A liquid fermentation and nattokinase technology, applied in the field of microorganisms, can solve the problems of industrial production limitation of nattokinase, low nattokinase enzyme activity, high price, etc., and achieves simple and easy extraction process, energy saving and raw material saving. , the effect of increasing NK production

Inactive Publication Date: 2011-02-23
HUBEI GOLE BIO TECH DEV
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Problems solved by technology

Different liquid fermentation methods of nattokinase are introduced respectively. The main raw material used is soybean peptone, and yeast extract and beef extract are used as raw materials for direct fermentation. The enzyme activity of nattokinase produced by these methods is not high, and the price is relatively expensive. Therefore, the industrialized production of nattokinase is subject to certain restrictions

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0017] 1. Mutation and domestication of nattokinase liquid fermentation strain

[0018] 1) The preparation method of bacterial suspension:

[0019] Inoculate the bacterium that has been activated on a fresh slant for 24 hours into the liquid seed medium, cultivate it at 37°C and 200r / min to the exponential growth phase, collect the bacterium by centrifugation, wash 2-3 times with sterilized normal saline, and transfer it into Shake in a Erlenmeyer flask filled with sterilized glass beads to make a cell concentration of 10 6 -10 8 cells / mL of bacterial suspension.

[0020] 2) Ultraviolet microwave mutagenesis:

[0021] The dilution of the seed solution is 100 cells / mL. On the ultra-clean workbench, take 5 mL of the bacterial suspension that is the concentration of the bacterial suspension at the time of treatment. Put the bacterial suspension on a sterile plate, place it on a magnetic stirrer, and adjust the Petri dish The distance to the UV lamp (40W) is 30cm. Turn on the...

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Abstract

The invention discloses a liquid fermentation method for producing natto kinase in high yield, which comprises a fermentation formula and fermentation conditions. In the method, a natto solid fermentation high-yield strain is selected and subjected to mutation and domestication so as to screen a high-yield strain of natto kinase; and the high-yield strain is inoculated into a liquid nutrient medium for fermentation, wherein the enzyme activity can reach 1,400IU / mL under liquid fermentation of a shake flask. Enzymatic hydrolysate of bean pulp is adopted as a nitrogen source of pilot plant test fermentation, and in a 10L fermentation tank, the fermentation unit can reach 2,650IU / mL. Due to the technical scheme, the yield of the prepared fermentation product natto kinase is high, the enzyme activity is high, the raw materials are saved, the energy is saved, the used raw materials are convenient to obtain and rich, the cost is low, and the extraction technology is easy and practicable.

Description

technical field [0001] The invention belongs to the technical field of microorganisms. The invention relates to the breeding of a high-yield nattokinase strain, soybean meal enzymatic hydrolysis liquid product and a liquid fermentation method of nattokinase. Background technique [0002] Thrombotic disease is a serious threat to human life and health, and its morbidity and mortality rank first among various diseases. Thrombolysis is the treatment of choice for this type of disease. At present, the clinical treatment of thrombosis is mainly through the injection of streptokinase (Streptokinase, SK), urokinase (Urokinase, UK), tissue-type plasminogen activator (tissue-type plasminogen activator, t-PA) and single-chain urokinase type Plasminogen activator (Single-chain Urokinase Type Plasminogen Activator, scu-PA), etc. These drugs are plasminogen activators, which degrade fibrin by activating plasminogen to generate plasmin. However, in practical applications, they all hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/56C12R1/125
Inventor 胡伶俐李军张云峰曹晓琴陈晓炜谢明利
Owner HUBEI GOLE BIO TECH DEV
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