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Gene related to proteolytic ability and screening method of lactobacillus helveticus with high proteolytic ability based on gene

A technology of proteolysis and lactobacillus, applied in the field of functional genes, achieves the effect of fast screening, simple method, and high-throughput screening results

Pending Publication Date: 2020-12-04
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is little research on the regulatory mechanism of Lactobacillus helveticus proteolytic activity, so there is no report on the screening of Lactobacillus helveticus with high proteolytic activity in terms of genes regulating proteolytic activity

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  • Gene related to proteolytic ability and screening method of lactobacillus helveticus with high proteolytic ability based on gene
  • Gene related to proteolytic ability and screening method of lactobacillus helveticus with high proteolytic ability based on gene
  • Gene related to proteolytic ability and screening method of lactobacillus helveticus with high proteolytic ability based on gene

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Experimental program
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Embodiment 1

[0058] 1. Materials and methods

[0059] 1.1 Isolation of strains

[0060] The 60 strains of Lactobacillus helveticus used in this study were isolated from fermented dairy products collected by our laboratory from Mongolia and several regions in China, including Inner Mongolia, Xinjiang, Gansu, Tibet and Yunnan, between 2002 and 2009 (Table 1).

[0061] Table 1. List of Lactobacillus helveticus strains used in this study

[0062]

[0063]

[0064] 2.2 Fermentation test and proteolytic activity of Lactobacillus helveticus

[0065] Preparation of fermented milk in fermentation bottles. Take 5×10 6 CFU / mL was inoculated with the test strain of Lactobacillus helveticus into fresh milk, and fermented at 37°C until the pH reached 4.5. The proteolytic activity of Lactobacillus helveticus was evaluated by measuring the content of free amino nitrogen in fermented milk at the end of fermentation (pH 4.5). The fermented milk was placed at 4°C and centrifuged at 6000×g for 10 m...

Embodiment 2

[0092] Screening of strains with high proteolytic ability

[0093] Lactobacillus helveticus samples whose proteolytic ability had been tested through the existing technology in the lactic acid bacteria resource bank of Inner Mongolia Agricultural University were used as the analysis objects, and a total of 20 strains were collected. See Table 4 for strain information.

[0094] Table 4 Proteolytic ability of Lactobacillus helveticus fermented to pH 4.5

[0095]

[0096] Do the following for each sample:

[0097] Bacterial strain activation: put the freeze-dried bacterial powder stored in the ampoule tube into the MRS medium for activation and rejuvenation (37°C, 24h), and microscopically examine the smear of the bacterial liquid, and the microscopic examination result is a Gram-positive pure culture The species were continued to be subcultured to the third generation.

[0098] Use liquid nitrogen freeze-thaw-CTAB method to extract strain DNA: wash the above-mentioned thir...

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Abstract

The invention provides a gene related to protein hydrolysis capacity and a screening method for lactobacillus helveticus with high protein hydrolysis capacity based on the gene, and belongs to the technical field of molecular biology. According to the invention, a whole-genome correlation analysis technology is adopted for rapid screening so as to obtain a coding hypothetical protein gene relatedto lactobacillus helveticus proteolytic activity. The hypothetical protein gene regulates and controls the proteolysis capacity of lactobacillus helveticus in a negative regulation and control mode, and can be used as a molecular marker for detecting the proteolysis activity of lactobacillus helveticus to be applied to screening of lactobacillus helveticus strains. The screening work of lactobacillus helveticus strains with high protein hydrolysis capacity is greatly simplified, and a new thought is provided for the strain screening work.

Description

technical field [0001] The invention belongs to the technical field of functional genes, and specifically relates to a gene related to proteolysis ability and a method for screening Lactobacillus helveticus with high proteolysis ability based on the gene. Background technique [0002] Lactobacillus helveticus (L.helveticus), as a lactic acid bacterium (Lactic Acid Bacteria, LAB), its excellent proteolytic activity can shorten the fermentation maturation time, promote the production of flavor substances, and reduce the bitterness of fermented milk (Fortina et al. ,1998), making it an important thermophilic industrial starter, widely used in the production of cheese, fermented milk and other foods (Sadat-Mekmene et al., 2011a). Lactobacillus helveticus releases a variety of important biologically active peptides by hydrolyzing milk proteins during the fermentation of dairy products (Beltran-Barrientos et al., 2016), among which the most studied angiotensin-converting enzyme (A...

Claims

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Application Information

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IPC IPC(8): C12N15/57C12N9/52C12N15/11C12Q1/689C12Q1/686C12Q1/6809C12Q1/04C12R1/225
CPCC12N9/52C12Y304/21096C12Q1/689C12Q1/686C12Q1/6809C12Q2565/125C12Q2535/122
Inventor 钟智赵洁扎木苏郭丽如陈永福孙志宏于洁刘文俊
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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