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Peptides natural microbial antiseptic agent producing strain, its use and preparation method for antiseptic agent

A technology of microbial preservatives and preservatives, applied in the direction of biochemical equipment and methods, botany equipment and methods, applications, etc., can solve the problems that the bacteria produced are under research, achieve the function of regulating the gastrointestinal tract, and have a wide range of antibacterial , the effect of treating allergic rhinitis

Inactive Publication Date: 2005-09-21
HEILONGJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many lactic acid bacteria can metabolize to produce peptides. Nisin is clearly produced by the metabolism of Streptococcus lactis. Many scholars have isolated a variety of lactic acid bacteria that produce preservatives. under study

Method used

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  • Peptides natural microbial antiseptic agent producing strain, its use and preparation method for antiseptic agent

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Experimental program
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specific Embodiment approach 1

[0027] Specific embodiment one: present embodiment isolates and identifies Lactobacillus paracasei HD1.7 like this:

[0028] ①The strains were isolated from L-lactic acid bacteria pickled sauerkraut fermentation broth by conventional plate streak separation method.

[0029] ②Screening of strains capable of producing antibacterial substances: After each single colony was expanded and cultivated on a plate, it was inserted into the fermentation broth for 48 hours at 37°C, and the fermentation broth was centrifuged at 3500r / min for 12 minutes. Staphylococcus aureus, Escherichia coli, Salmonella and other 12 kinds of indicator bacteria were cultured on a double-layer detection plate for 12 to 24 hours, and the antibacterial effect was observed, and the strains with broad antibacterial spectrum and strong antibacterial ability were retained.

[0030] ③ A strain that can metabolize and produce peptide preservatives was obtained, which was identified as Lactobacillus paracasei (Lacto...

specific Embodiment approach

[0031] The specific implementation method is as follows: (1) using conventional culture characteristics, physiological and biochemical characteristics to identify peptide preservative producing bacteria as Lactobacillus; Design PCR primers based on the conserved sequences at both ends of the 16SrDNA gene of many species of Lactobacillus, extract DNA by lysozyme-SDS-potassium acetate method, amplify by PCR, electrophoresis, recover PCR products, and clone PCR products into plasmid vectors, After being amplified in the bacteria, the carrier with the 16SrDNA gene was used as a template, and M13F or M13R was used as a primer for sequencing, and the 16SrDNA sequence analysis was carried out to identify the peptide preservative producing bacteria as Lactobacillus paracasei HD1.7 (Lactobacillus paracasei HD1.7 ).

specific Embodiment approach 2

[0032] Specific embodiment two: the preparation method of the peptide natural microbial preservative of the present embodiment includes the following steps: a, fermentation of the peptide preservative: the preservation slant of Lactobacillus paracasei HD1.7 is streaked and inoculated on the fermentation medium, 35-40°C upside-down culture for 36-48 hours, then inoculated into the seed fermentation liquid, and then cultured statically at 35-40°C for 24-36 hours, then inoculated with 1-3% of the inoculation amount into the expanded culture fermentation liquid, at 35-40 Cultivate statically for 24-36 hours at ℃, then inoculate in the secondary expansion culture fermentation broth with 1-3% inoculum amount, culture statically at 35-40°C for 24-36 hours, continue to inoculate with 1-3% inoculum amount In the fermentation broth, culture statically at 35-40°C for 30-50 hours; b. Separation and purification of peptide preservatives: centrifuge the fermentation broth of Lactobacillus pa...

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Abstract

This invention relates to an antiseptic producing bacteria, its metabolite application and preparing natural microbios antiseptic process using the metabolite as material. Naming the invention as Lactobacillus paracaseiHD1.7, preservation number being CCTCC: M205015. Its metabolite is a kind of peptide antiseptic, being capable of inhibiting colibacillus, Staphylococcus aureus, micrococcus flavus, bacillus subtilis, also salmonella, bacillus megaterium, Bacillus cereus, Shigella, proteus and such like these pathogenic bacteria; in condition of 120Deg.C, 60min still sustaining its bacterial inhibition active, with strong heat stability; being irresponsive to several proteinases, working well in neutral or acidic conditions.

Description

Technical field: [0001] The invention relates to the isolation and identification of peptide preservative-producing bacteria, the application of the metabolites of the preservative-producing bacteria, and a method for preparing natural microbial preservatives using the metabolites of the preservative-producing bacteria as raw materials. Background technique: [0002] Preservatives are divided into chemical preservatives and natural microbial preservatives. At present, the preservatives widely used in the food industry at home and abroad are still synthesized by artificial chemical methods. The raw materials, auxiliary materials and additives used in the production process or the impurities contained in them may cause harm to human health; many chemical preservatives are not Substances that exist naturally in nature, whether they have potential chronic toxicity or tertiary effects on human health are not completely clear. At present, there are only a few natural microbial pr...

Claims

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Application Information

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IPC IPC(8): C12NC12N1/20C12N15/31C12P1/04
Inventor 平文祥周东坡雷虹李秀凉张钦革王建臣贾树彪
Owner HEILONGJIANG UNIV
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