Preparation method of diet-fermented konjak juice rich in pyrroloquinoline quinone and nattokinase

A technology of pyrroloquinoline quinone and nattokinase, which is applied in the field of microbial engineering and fermentation, and can solve the problems of unrelated breeding medium and fermentation medium, low yield of target products and the like

Inactive Publication Date: 2015-03-25
HANGZHOU YUANPEITE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 diet-fermented konjak juice rich in pyrroloquinoline quinone and nattokinase
  • Preparation method of diet-fermented konjak juice rich in pyrroloquinoline quinone and nattokinase
  • Preparation method of diet-fermented konjak juice rich in pyrroloquinoline quinone and nattokinase

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preparation example Construction

[0103] (h) The preparation of konjac juice includes: adding 190 g of konjac cubes to 1000 mL of water, soaking for 10 hours, and grinding at 95° C. to obtain konjac juice.

[0104] 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, purchased from Jiaxing Yua...

Embodiment 1

[0106] The present invention prepares konjac juice rich in NK and PQQ through the breeding of Bacillus natto and Lactobacillus casei and their co-fermentation, comprising the following steps:

[0107] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-304 with high yield of NK and PQQ

[0108] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-304 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 as a p...

Embodiment 2

[0120] The present invention prepares konjac juice rich in NK and PQQ through the breeding of Bacillus natto and Lactobacillus casei and their co-fermentation, comprising the following steps:

[0121] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-304 with high yield of NK and PQQ

[0122] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-304 was obtained through genome rearrangement and high-throughput screening technology. The obtained NK and PQQ-producing Bacillus natto 10261 and 10263 were subjected to nitrosoguanidine mutagenesis respectively; then the offspring were mixed for mutagenesis, 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 suitable as ...

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Abstract

The invention discloses a preparation method of diet-fermented konjak juice rich in pyrroloquinoline quinone and nattokinase. The preparation method comprises the following steps: respectively carrying out nitrosoguanidine mutagenesis on bacillus natto 10261 and 10263 producing nattokinase and pyrroloquinoline quinone, then mixing mutagenic progenies of nattokinase and pyrroloquinoline quinone for cell fusion subculture, and based on a genome rearrangement technology, further adopting a substrate induction adaptive strategy and performing high throughput screening to obtain bacillus natto highly yielding nattokinase and pyrroloquinoline quinone; carrying out nitrosoguanidine induction on lactobacillus casei and screening lactobacillus casei lack of methionine nutrition; and co-fermenting konjak juice by the screened bacillus natto and lactobacillus casei to obtain konjak juice rich in pyrroloquinoline quinone and nattokinase. In the konjak juice rich in pyrroloquinoline quinone and nattokinase disclosed by the invention, the content of pyrroloquinoline quinone reaches 68-93ng/mL and the activity of the nattokinase reaches 392-766U/mL.

Description

technical field [0001] The invention relates to the field of microbial engineering and fermentation, in particular to a method for preparing dietary fermented konjac 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. Linolei...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A23L2/02C12N15/03C12N15/01C12R1/07C12R1/245
CPCA23L2/382C12N15/01C12N15/03
Inventor 吴渊徐娟李青青
Owner HANGZHOU YUANPEITE BIOTECH
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