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Method for continuously extracting nattokinase and gamma-polyglutamic acid from natto

A technology of nattokinase and polyglutamic acid, applied in the field of microbial engineering, to achieve high extraction rate, efficient utilization, and simple operation

Pending Publication Date: 2022-03-01
SHANGHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on the continuous extraction of nattokinase and γ-polyglutamic acid

Method used

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  • Method for continuously extracting nattokinase and gamma-polyglutamic acid from natto

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Experimental program
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Effect test

Embodiment 1

[0028] A method for continuously extracting nattokinase and gamma-polyglutamic acid, the technological process is as follows figure 1 shown, including the following steps:

[0029] (1) Preparation of medium: first prepare the medium of Bacillus subtilis, slant medium: LB medium 40g, agar 20g, distilled water 1L, natural pH, sterilized at 121°C for 20min; liquid medium: sucrose 30g, beef extract 10g, sodium glutamate 30g, MgSO 4 ·7H 2 O 0.25g, K 2 HPO 4 0.5g, CaCl 2 1g, natural pH, 1L of distilled water, sterilized at 121°C for 15 minutes; solid fermentation medium: 10g of soybeans, washed and soaked for 8 hours, drained and steamed and sterilized at 121°C for 20 minutes.

[0030] (2) Activation culture of Bacillus subtilis: Inoculate Bacillus subtilis stored at 4°C into slant culture medium under aseptic conditions, cultivate at 37°C for 24 hours to recover the strain, and then pick seeds and inoculate them into a medium containing 50mL of liquid medium. In a shaking f...

Embodiment 2

[0039] A method for continuously extracting nattokinase and gamma-polyglutamic acid, the technological process is as follows figure 1 shown, including the following steps:

[0040] (1) Preparation of culture medium: first prepare the culture medium of Bacillus subtilis, slant medium: 40g of LB medium, 20g of agar, 1L of distilled water, natural pH, sterilized at 121°C for 20min; liquid medium: 30g of sucrose, beef extract 10g, sodium glutamate 30g, MgSO 4 ·7H 2 O 0.25g, K 2 HPO 4 0.5g, CaCl 2 1g, natural pH, 1L of distilled water, sterilized at 121°C for 15 minutes; solid fermentation medium: 10g of soybeans, washed and soaked for 8 hours, drained and steamed and sterilized at 121°C for 20 minutes.

[0041] (2) Activation culture of Bacillus subtilis: Inoculate Bacillus subtilis stored at 4°C into slant culture medium under aseptic conditions, cultivate at 37°C for 24 hours to recover the strain, and then pick seeds and inoculate them into a medium containing 50mL of li...

Embodiment 3

[0050] A method for continuously extracting nattokinase and gamma-polyglutamic acid, the technological process is as follows figure 1 shown, including the following steps:

[0051] (1) Preparation of culture medium: first prepare the culture medium of Bacillus subtilis, slant medium: 40g of LB medium, 20g of agar, 1L of distilled water, natural pH, sterilized at 121°C for 20min; liquid medium: 30g of sucrose, beef extract 10g, sodium glutamate 30g, MgSO 4 ·7H 2 O 0.25g, K 2 HPO 4 0.5g, CaCl 2 1g, natural pH, 1L of distilled water, sterilized at 121°C for 20 minutes; solid fermentation medium: 10g of soybeans, washed and soaked for 8 hours, drained and steamed and sterilized at 121°C for 20 minutes.

[0052] (2) Activation culture of Bacillus subtilis: Inoculate Bacillus subtilis stored at 4°C into slant culture medium under aseptic conditions, cultivate at 37°C for 24 hours to recover the strain, and then pick seeds and inoculate them into a medium containing 50mL of li...

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Abstract

The invention discloses a method for continuously extracting nattokinase and gamma-polyglutamic acid from natto. The method comprises the following steps: firstly, carrying out recovery and activation culture on bacillus subtilis to obtain a bacillus subtilis seed solution; preparing a bacillus subtilis fermented solid soybean culture medium, inoculating the activated bacillus subtilis seed solution, and fermenting to obtain a fermentation product; and separating the fermentation product by using a salting-out method and an organic reagent precipitation method in sequence, and extracting to obtain nattokinase and gamma-polyglutamic acid. Nattokinase is an antithrombotic drug with great potential at present, and gamma-polyglutamic acid can be widely applied to the fields of agriculture, food, medicine, cosmetics and the like. The method is high in extraction rate, the required reagent is safe and cheap, the operability is high, and the fermentation efficiency and value of the soybeans can be effectively improved.

Description

technical field [0001] The invention relates to a method for continuously extracting nattokinase and gamma-polyglutamic acid from natto, belonging to the technical field of microbial engineering. Background technique [0002] The fermentation product of solid fermented soybeans with Bacillus subtilis is called natto, which contains more nattokinase (NK for short) and γ-polyglutamic acid (γ-PGA). Nattokinase is a serine protease and the main physiologically active component of natto. Compared with other thrombolytic agents such as urokinase and streptokinase, nattokinase has the advantages of less gastrointestinal side effects, non-toxic and harmless, no internal bleeding, good stability, and long half-life. It can not only activate plasminogen in the human body to convert it into plasmin, but also has a strong thrombolytic ability. Further clinical studies have shown that nattokinase can also significantly inhibit and improve platelet aggregation and hypertension. [0003...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/54C12P13/02C12R1/125
CPCC12N1/20C12N9/54C12P13/02C12Y304/21062
Inventor 马霞刘芷含何艳
Owner SHANGHAI INST OF TECH