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Preparation method of sticky rice juice containing rich nattokinase and pyrroloquinoline quinine

A technology of pyrroloquinoline quinone and nattokinase, which is applied in the directions of microorganism-based methods, biochemical equipment and methods, and food preparation, etc. question

Inactive Publication Date: 2015-04-22
HANGZHOU YUANPEITE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0025] However, there are certain defects in the strategy of genome rearrangement at present, mainly because there is no connection between the breeding medium and the fermentation medium, so that the yield of the target product of the selected strains during fermentation is much lower than expected

Method used

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  • Preparation method of sticky rice juice containing rich nattokinase and pyrroloquinoline quinine
  • Preparation method of sticky rice juice containing rich nattokinase and pyrroloquinoline quinine
  • Preparation method of sticky rice juice containing rich nattokinase and pyrroloquinoline quinine

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preparation example Construction

[0106] (h) The preparation of glutinous rice juice includes: adding 190 g of glutinous rice to 1000 mL of water, soaking for 6 hours, and grinding at a temperature above 95° C. to form a slurry juice.

[0107] In the embodiment of the present invention, Bacillus natto 10261 and Bacillus natto 10263 adopt the prior art, and Bacillus natto 10261 comes from China Industrial Microorganism Culture Collection Center (CICC), address: 24 Jiuxianqiao Middle Road, Chaoyang District, Beijing No. 6 building; the storage date is 2002, and the strain maintenance number is 10261; Bacillus natto 10263 is from the China Industrial Microbiology Culture Collection Center (CICC), address: Courtyard 6, No. 24, Jiuxianqiao Middle Road, Chaoyang District, Beijing Building No. 1; the storage date was 2002, and the strain maintenance number was 10263; Lactobacillus casei Shirota was from Yakult (Shanghai) Co., Ltd.; the mouse anti-PQQ IgG monoclonal antibody PQQ-mAb was a commercially available product...

Embodiment 1

[0109] The present invention prepares glutinous rice juice rich in NK and PQQ through breeding of Bacillus natto and Lactobacillus casei and co-fermentation thereof, comprising the following steps:

[0110] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-309 with high yield of NK and PQQ

[0111] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-309 was obtained through genome rearrangement and high-throughput screening technology. The obtained Bacillus natto 10261 and 10263 producing NK and PQQ were subjected to nitrosoguanidine mutagenesis respectively; then the offspring were mixed for mutagenesis, and cell fusion was carried out (fusogenic agent was polyethylene glycol 4000), subcultured, Realized genome shuffling, optimized and integrated the dominant mutations of all mutagenized progenies of Bacillus natto 10261 and 10623, and bred a high-yield NK and PQQ Natto with high yield, vigorous growth, stable genetics, and suitable ...

Embodiment 2

[0123] The present invention prepares glutinous rice juice rich in NK and PQQ through breeding of Bacillus natto and Lactobacillus casei and co-fermentation thereof, comprising the following steps:

[0124] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-309 with high yield of NK and PQQ

[0125] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-309 was obtained through genome rearrangement and high-throughput screening technology. The obtained Bacillus natto 10261 and 10263 producing NK and PQQ were respectively subjected to nitrosoguanidine mutagenesis; then the offspring were mixed for mutagenesis, and cytoplasmic fusion was carried out (fusogenic agent was polyethylene glycol 4000), subcultured, Realized genome shuffling, optimized and integrated the dominant mutations of all mutagenized progenies of Bacillus natto 10261 and 10623, and bred a high-yield NK and PQQ Natto with high yield, vigorous growth, stable genetics, and su...

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Abstract

The invention discloses a preparation method of sticky rice juice containing rich nattokinase and pyrroloquinoline quinine. The preparation method comprises the steps of carrying out nitrosoguanidine mutation on bacillus natto 10261 and 10263 respectively, wherein the bacillus natto 10261 and 10263 produce nattokinase and pyrroloquinoline quinine, mixing the mutation progenies of the bacillus natto 10261 and 10263, carrying out cell fusion subculture, further adopting the substrate-induced adaptable strategy on the basis of the genome rearrangement technology, and performing high-throughput screening to obtain the bacillus natto which has high yield of nattokinase and pyrroloquinoline quinine; carrying out nitrosoguanidine mutation on Lactobacillus casei and screening out methionine auxotrophic Lactobacillus casei; and co-fermenting sticky rice juice by use of the screened bacillus natto and the Lactobacillus casei to obtain the sticky rice juice containing rich nattokinase and pyrroloquinoline quinine. According to the sticky rice juice containing rich nattokinase and pyrroloquinoline quinine, the content of the pyrroloquinoline quinine is up to 67-118ng / mL and the activity of the nattokinase can reach 472-780U / mL.

Description

technical field [0001] The invention relates to the field of microbial engineering and fermentation, in particular to a method for preparing glutinous rice juice rich in nattokinase and pyrroloquinoline quinone. Background technique [0002] Natto is fermented from soybeans by Bacillus natto, and has multiple physiological functions: [0003] ①The effect of natto on the cardiovascular system: Nattokinase (Nattokinase, NK), a symbolic physiologically active substance in natto, is a protein capable of dissolving thrombus. It consists of 275 amino acid residues and has a molecular weight of about 27KD. It is non-toxic, It has no side effects and is a serine protease whose thrombolytic effect even exceeds that of the thrombolytic agent urokinase. In addition to its direct thrombolytic effect, nattokinase also has the ability to promote the production of plasminogen activator in venous endothelial cells, thereby indirectly expressing its thrombolytic activity. Linoleic acid in ...

Claims

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Application Information

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IPC IPC(8): A23L2/38A23L1/29C12N15/03C12N15/01C12R1/07C12R1/245A23L33/00
CPCA23L2/382C12N15/01C12N15/03
Inventor 徐娟吴渊邓惟
Owner HANGZHOU YUANPEITE BIOTECH
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