Method for preparing nattokinase through solid-state fermentation

A technology of nattokinase and solid-state fermentation, applied in the field of microbial fermentation engineering, can solve the problem that nattokinase has not been reported, and achieve the effects of high nattokinase activity, increased added value, and convenient operation.

Active Publication Date: 2016-11-09
烟台泰禾电子科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Liu Hongmei and others prepared peanut antioxidant peptides by liquid fermentation of four microorganisms including Bacillus subtilis AS1.398, Bacillus licheniformis 2709 and Lactobacillus plantarum, but there is no report on the preparation of nattokinase by fermenting peanut meal with Bacillus natto

Method used

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  • Method for preparing nattokinase through solid-state fermentation
  • Method for preparing nattokinase through solid-state fermentation
  • Method for preparing nattokinase through solid-state fermentation

Examples

Experimental program
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Effect test

Embodiment 1

[0018] The concrete process that present embodiment prepares nattokinase is:

[0019] (1) Preparation of medium: Mix agar 15g / L, peptone 10g / L, beef powder 3g / L and sodium chloride 5g / L, and then sterilize at 121°C for 20 minutes to obtain a slant medium with a pH of 7.1 to 7.5 ; Mix peptone 10g / L, beef extract 5g / L, sodium chloride 3g / L, glucose 10g / L and yeast extract 5g / L, and then sterilize at 121°C for 20min to obtain a seed liquid medium; peanut meal, Wash the bran, soybean meal, oats, and soybeans with clean water and soak them in water three times the total weight, then mix them in a weight ratio of 6:1:1:1:1, and add peanut meal, bran, soybean meal, oats and A nutrient salt composed of a mixture of 1.5% sucrose, 0.21% magnesium sulfate, and 0.27% calcium chloride of the total weight of soybeans, mixed evenly and sterilized at 121° C. for 20 minutes to obtain a fermentation medium;

[0020] (2) Preparation of Bacillus natto seed solution: in a sterile environment, pic...

Embodiment 2

[0023] In this embodiment, the fermented seed solution prepared in step (2) of Example 1 is sampled every 3 hours, and the OD value at 660 nm is measured, and the growth of Bacillus natto is plotted with the seed culture solution not inoculated with Bacillus natto as a blank control curve (eg figure 1 shown), determine the seed age of the access fermentation medium; and change the fermentation temperature, inoculum size (spraying amount of fermented seed liquid), fermentation time, material-liquid ratio (fermentation medium and the Water weight ratio) and the nutrient salt in the step (1), measure the influence of single factor on nattokinase activity and soluble nitrogen content respectively, fermentation temperature is respectively 29,33,37,41,45 ℃; Inoculum size is respectively 1, 2, 3, 4, 5mL; the fermentation time is 12, 18, 24, 30, 36h; the ratio of solid to liquid is 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6 The measurement results are as image 3 , 4 , 5, and 6; Nutrient sa...

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Abstract

The invention belongs to the technical field of microorganism fermentation engineering and relates to a method for preparing nattokinase through solid-state fermentation. The method includes: respectively preparing a bevel culture medium, a seed solution culture medium and a fermentation culture medium; inoculating bacillus natto onto the bevel culture medium for culture to obtain activated bacillus natto, picking and placing 1-2 rings into the seed solution culture medium for oscillation culture to serve as a fermentation seed solution; sterilizing and then naturally cooling the fermentation culture medium, spraying the fermentation seed solution into the fermentation culture medium, adding water, and stirring well; paving a thin layer in a beaker, using gauze to seal, sequentially performing fermentation and after-ripening for 24 h, and adding normal saline; performing extraction and centrifuging to obtain supernate rich in nattokinase. The method is simple in process and convenient to operate, raw materials are easy to get and low in cost, and nattokinase prepared by the method is high in activity and has obvious economic benefit.

Description

Technical field: [0001] The invention belongs to the technical field of microbial fermentation engineering, and relates to a technique for preparing nattokinase by solid-state fermentation using peanut meal as a raw material, in particular to a method for preparing nattokinase by solid-state fermentation. Background technique: [0002] Thrombosis is formed by the aggregation of fibrinogen and platelets, which often causes a series of diseases and is a cardiovascular disease that seriously endangers human health. Nattokinase (nattokinase referred to as NK, is a serine protease produced by Bacillus subtilisl natto), nattokinase has been proved to be an ideal drug candidate for the treatment of cardiovascular and cerebrovascular diseases, and other traditional thrombolytic drugs Compared with nattokinase, it has many advantages such as good safety, low cost, easy absorption by human body, direct and rapid action, and long duration of action. Therefore, nattokinase, as a new gen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/54C12R1/07
CPCC12N9/54
Inventor 胡迎芬王艳平马爱国梁惠张源洁刘青丁皓玥
Owner 烟台泰禾电子科技有限公司
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