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Weakly post-acidified Lactobacillus helveticus sh2-5-66 and its application

A post-acidification, lactobacillus technology, applied in the field of lactic acid bacteria, can solve the problems of complicated operation, retention of lactic acid bacteria, unstable genetic properties and other problems

Active Publication Date: 2021-09-10
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the modification of lactic acid bacteria in China is still at the level of artificial mutagenesis, the operation is complicated, and the obtained traits cannot be stably inherited

Method used

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  • Weakly post-acidified Lactobacillus helveticus sh2-5-66 and its application
  • Weakly post-acidified Lactobacillus helveticus sh2-5-66 and its application
  • Weakly post-acidified Lactobacillus helveticus sh2-5-66 and its application

Examples

Experimental program
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Effect test

preparation example Construction

[0059] (1) Preparation of Fermented Milk Crude Enzyme Liquid

[0060] Take 10.0 g of fermented milk for the 1st, 5th, 10th, 15th, and 20th day of storage, remove the protein with 1% EDTA solution (pH=12, 4°C) and resuspend in TE buffer, add lysozyme, and bathe in 37°C for 1 hour. Centrifuge to remove the supernatant, resuspend with normal saline, and crush the cell suspension with a high-pressure cytometer to extract the crude enzyme solution for the determination of β-galactosidase and H + - ATPase enzyme activity.

[0061] (2) Determination of β-galactosidase activity

[0062] (a) draw ONP (o-nitrophenol) standard curve

[0063] (b) β-galactosidase activity assay

[0064] Take 1mL of crude enzyme solution, put it in a water bath at 37°C for 10min, add 1mL of ONPG solution (pH 7.0, 20mmol / L), react at 37°C for 10min, and immediately add 3mL of 0.5mol / L Na 2 CO 3 The solution terminated the reaction, and the absorbance was measured at 420 nm. .

[0065] β-galactosidase ...

Embodiment 1

[0082] Mutation Breeding, Screening and Identification of Weak Postacidification Strains

[0083] 1. Mutation breeding

[0084] Using a 20W ultraviolet lamp, the irradiation distance was 30cm, and the irradiation distance was 0s, 30s, 60s, 90s, and 120s respectively, and the lethal rate under the mutagenesis time was determined (Table 1). The lethality rate of the strain was over 80% after irradiating for 50s, so the time of mutagenic irradiation was set at 50s-60s.

[0085] Table 1 Effect of mutagenesis time on the lethality of L.helveticus SH2-1

[0086]

[0087] 2. Screening

[0088] (1) Establishment of post-acidification screening indicators

[0089] In order to determine the screening index of the enzyme coupled with post-acidification of Lactobacillus helveticus SH2-1 for the screening of late-stage mutagenesis strains, this part of the experiment measured the storage period index of L. helveticus SH2-1 after fermentation, and selected two strains with weak post-a...

Embodiment 2

[0102] (1) Fermentation effect experiment of L.helveticus sh2-5-66

[0103] Ferment SH2-1 (L. helveticus SH2-1) and sh2-5-66 (L. helveticus sh2-5-66) in a single strain, or mix it with commercial starter st447 (S. thermophilus st447) in different proportions Mixed bacteria fermentation, determination of the quality index of fermented milk, the specific experimental method is as follows:

[0104] Single-bacteria fermentation: Inoculate the activated strain into the prepared skim milk medium according to the amount of 3%, place it in a constant temperature incubator at 42°C for cultivation, observe the curdling situation, record the curdling time, and place it in a refrigerator at 4°C storage.

[0105] Mixed bacteria fermentation: compound st447 with Lactobacillus helveticus SH2-1 and sh2-5-66 before and after mutagenesis according to the ratio of 1:1, 1.5:1, and 2:1, and the total number of viable bacteria added is 6 ×10 6 cfu / mL for inoculation and fermentation, placed in a...

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Abstract

The invention discloses a strain of Lactobacillus helveticus sh2‑5‑66 and its application. In the present invention, human Lactobacillus helveticus SH2‑1 with excellent probiotic properties is used as the starting strain, through ultraviolet mutagenesis, and according to H + Relationship between ‑ATPase enzyme and acid production of Lactobacillus helveticus SH2‑1 during storage, expressed as H + ‑ATPase enzyme activity was used as a screening index, and the weak postacidified Lactobacillus helveticus sh2‑5‑66 was obtained, and the preservation number was CGMCC No.19123. Utilize Lactobacillus helveticus sh2-5-66 of the present invention to carry out fermented milk production, compared with Lactobacillus helveticus SH2-1, acidity reduces 57 ° T, utilizes Lactobacillus helveticus sh2-5-66 alone or with commercial thermophilic chain The fermented milk produced by coccus st447 has excellent fermentation indicators, and Lactobacillus helveticus sh2‑5‑66 has excellent genetic stability.

Description

technical field [0001] The invention belongs to the technical field of lactic acid bacteria, and relates to a weak post-acidification Lactobacillus helveticus sh2-5-66 and its application. Background technique [0002] Fermented milk is a milk product with special flavor and rich nutrition, which is made from milk or milk powder, which is sterilized and then fermented with lactic acid bacteria. It is favored by consumers because of its functions of promoting digestion, alleviating lactose intolerance, regulating the balance of intestinal flora, and lowering cholesterol. After the fermented milk is fermented, it needs to be refrigerated at a temperature of 2-4°C for 12-24 hours, which is called the post-ripening period or post-acidification of fermented milk. The minimum pH of fermented milk acceptable to consumers is generally 4.2. However, in the low temperature environment before transportation and sales, lactic acid bacteria still have certain activity and will continue...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23C9/123A23C19/00A23L13/40A23L13/60C12R1/225C12R1/46
CPCC12N1/20A23C9/123A23C19/00A23L13/65A23L13/45C12R2001/225C12N1/205A23V2400/147
Inventor 关成冉陈萱陶志强王丽赵瑞锋陈大卫张臣臣黄玉军陈霞顾瑞霞
Owner YANGZHOU UNIV
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