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Method for preparing dietary therapy orange juice by virtue of breeding and co-fermentation of bacillus natto and lactobacillus casei

A technology of Bacillus natto and Lactobacillus casei, applied in Lactobacillus, microorganism-based methods, biochemical equipment and methods, etc., can solve the problem of low yield of target products, unrelated breeding medium and fermentation medium, etc. question

Inactive Publication Date: 2015-03-25
HANGZHOU YUANPEITE BIOTECH
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  • 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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  • Method for preparing dietary therapy orange juice by virtue of breeding and co-fermentation of bacillus natto and lactobacillus casei
  • Method for preparing dietary therapy orange juice by virtue of breeding and co-fermentation of bacillus natto and lactobacillus casei
  • Method for preparing dietary therapy orange juice by virtue of breeding and co-fermentation of bacillus natto and lactobacillus casei

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preparation example Construction

[0101] (f) The preparation of the orange juice comprises: adding 200 g of orange in 1000 mL of water, and grinding it into pulp juice.

[0102] 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 Yuanke Biotechnology Co., Ltd., the prod...

Embodiment 1

[0104] The present invention prepares orange juice rich in NK and PQQ through the breeding of Bacillus natto and Lactobacillus casei and their co-fermentation, comprising the following steps:

[0105] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-501 with high yield of NK and PQQ

[0106] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-501 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

[0118] The present invention prepares orange juice rich in NK and PQQ through the breeding of Bacillus natto and Lactobacillus casei and their co-fermentation, comprising the following steps:

[0119] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-501 with high yield of NK and PQQ

[0120] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-501 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 suitable...

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Abstract

The invention discloses a method for preparing dietary therapy orange juice by virtue of breeding and co-fermentation of bacillus natto and lactobacillus casei. The method comprises the following steps: carrying out nitrosoguanidine mutagenesis respectively on bacillus natto 10261 and 10263 for producing nattokinase and pyrroloquinoline quinine, then mixing the two mutagenesis progenies, implementing cell fusion and subculture, and conducting high-throughput screening by further adopting a substrate-induced adaptive strategy on the basis of the genome shuffling technology so as to obtain bacillus natto for the high production of nattokinase and pyrroloquinoline quinine; carrying out nitrosoguanidine mutagenesis on lactobacillus casei and screening out methionine auxotrophic lactobacillus casei; and co-fermenting orange juice by virtue of the screened bacillus natto and lactobacillus casei so as to obtain the orange juice rich in nattokinase and pyrroloquinoline quinine. In the orange juice rich in nattokinase and pyrroloquinoline quinine prepared by the method disclosed by invention, the content of the pyrroloquinoline quinine reaches 50-95ng / mL and the activity of the nattokinase reaches 366-780U / mL.

Description

technical field [0001] The invention relates to the field of microbial engineering and fermentation, in particular to a method for co-fermenting bacillus natto and Lactobacillus casei to prepare therapeutic orange juice. 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...

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
CPCA23L2/02A23L2/84A23L33/00C12N15/01C12N15/03A23V2002/00A23V2400/125A23V2200/30
Inventor 阮子琦张哲滔吴渊莫凡
Owner HANGZHOU YUANPEITE BIOTECH
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