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Method of preparing yoghourt rich in nattokinase and pyrroloquinoline quinine through breeding bacillus natto and lactobacillus casei and co-fermentation

A technology of Bacillus natto and Lactobacillus casei, which is applied in the fields of dairy products, mutant preparation, hybrid cell preparation, etc., and can solve the problems of low yield of target products, unrelated breeding medium and fermentation medium, etc.

Inactive Publication Date: 2015-04-01
ZHEJIANG UNIV
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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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  • Method of preparing yoghourt rich in nattokinase and pyrroloquinoline quinine through breeding bacillus natto and lactobacillus casei and co-fermentation
  • Method of preparing yoghourt rich in nattokinase and pyrroloquinoline quinine through breeding bacillus natto and lactobacillus casei and co-fermentation
  • Method of preparing yoghourt rich in nattokinase and pyrroloquinoline quinine through breeding bacillus natto and lactobacillus casei and co-fermentation

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Embodiment 1

[0105] The present invention prepares yoghurt rich in NK and PQQ through the selection and co-fermentation of Bacillus natto and Lactobacillus casei, comprising the following steps:

[0106] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-1 with high yield of NK and PQQ

[0107] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-1 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 production stra...

Embodiment 2

[0119] The present invention prepares yoghurt rich in NK and PQQ through the selection and co-fermentation of Bacillus natto and Lactobacillus casei, comprising the following steps:

[0120] (1) Breeding of Bacillus natto BN-NKPQQ-RWX-1 with high yield of NK and PQQ

[0121] Using the substrate-induced adaptive strategy, the high-yield NK and PQQ Bacillus natto BN-NKPQQ-RWX-1 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 as a producti...

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Abstract

The invention discloses a method of preparing yoghourt rich in nattokinase and pyrroloquinoline quinine through breeding bacillus natto and lactobacillus casei and co-fermentation. The method comprises the following steps: carrying out nitrosoguanidine mutagenesis on bacillus natto 10261 and 10263 capable of producing nattokinase and pyrroloquinoline quinine respectively, then mixing the mutation progenies of the bacillus natto 10261 and 10263, and carrying out cell fusion subculture and high-throughput screening to obtain bacillus natto capable of producing nattokinase and pyrroloquinoline quinine with high yield; carrying out nitrosoguanidine mutagenesis on lactobacillus casei, and screening methionine auxotroph lactobacillus casei from the lactobacillus casei; co-fermenting fresh milk by using the screened bacillus natto and lactobacillus casei to obtain the yoghourt rich in nattokinase and pyrroloquinoline quinine. In the yoghourt rich in nattokinase and pyrroloquinoline quinine, which is obtained by the method disclosed by the invention, the content of pyrroloquinoline quinine reaches 137-149ng / mL, and the activity of nattokinase reaches 1935-2100U / mL.

Description

technical field [0001] The invention relates to the field of microbial engineering and fermentation, in particular to a method for breeding Bacillus natto and Lactobacillus casei and their co-fermentation to prepare yoghurt rich in nattokinase (NK) and pyrroloquinoline quinone (PQQ). 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, th...

Claims

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Application Information

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IPC IPC(8): A23C9/127C12N15/03C12N15/01
Inventor 阮晖徐薇薇朱梦娇张哲滔胡慧雯林永华莫凡
Owner ZHEJIANG UNIV
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