Separating, screening and identifying method of lactobacillus sake

A Lactobacillus sake, separation and screening technology, applied in the directions of microorganism-based methods, separation of microorganisms, biochemical equipment and methods, etc., can solve the problems of cumbersome preparation steps, affecting use and promotion, and single acquisition method, and achieve a reasonable operation process. and simple effect

Inactive Publication Date: 2017-01-04
JILIN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a preparation method of Lactobacillus sake, to solve the problems of its sin

Method used

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  • Separating, screening and identifying method of lactobacillus sake
  • Separating, screening and identifying method of lactobacillus sake
  • Separating, screening and identifying method of lactobacillus sake

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Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0019] Specific implementation mode one: this implementation mode is carried out according to the following steps:

[0020] Step 1: Preliminary screening of bacterial strains: firstly, select multiple cooked meat samples of equal weight, and each cooked meat sample corresponds to a portion of normal saline, and at the same time, divide each selected cooked meat sample in a mass ratio of 1:9 The proportion of the corresponding physiological saline is diluted to form a variety of bacterial liquids, and then the various bacterial liquids are sequentially inoculated into the cells containing CaCO 3 In the MRS solid medium, screen out multiple strains producing transparent circles, and then perform single colony culture on multiple strains one by one, and pick multiple single colonies for H 2 o 2 Enzyme contact experiment and Gram staining experiment, finally choose H 2 o 2 Multiple strains that are negative for enzyme contact and positive for Gram staining are multiple strains ...

specific Embodiment approach 2

[0032] Specific embodiment two: this embodiment is a further limitation of specific embodiment one, and the operation process of the antibacterial test in step 2 in this embodiment is:

[0033] Add 10mL of MRS medium at the bottom of the plate, pour the Oxford cup into the plate, pour in 15mL of LB medium after cooling the medium, take off the Oxford cup after the LB medium is cooled, and inject 100μL of the liquid medium supernatant, The diameter of the inhibition zone after 24 hours of culture.

[0034] In this embodiment, both the MRS medium and the LB medium are existing medium.

specific Embodiment approach 3

[0035] Embodiment 3: This embodiment is a further limitation of Embodiment 1. In this embodiment, the Gram-positive bacteria in Step 2 are Staphylococcus aureus; the Gram-negative bacteria are Escherichia coli.

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Abstract

Lactobacillus sake is a natural food preservative and feed additive with potential development capacity; the lactobacillus sake is simplex in obtaining method and small in yield, so use and popularization of the lactobacillus sake are seriously influenced; meanwhile the lactobacillus sake is complex in preparation step and high in expenditure. A method for obtaining the lactobacillus sake by separating, screening and identifying bacteriocin producing lactic acid bacteria includes the steps of firstly, primary screening work of strains; secondly, secondary screening work of the strains, wherein in the secondary screening of the strains, an antibacterial test is conducted on a plurality of lactic acid bacteria strains obtained in the first step with gram-positive bacteria and gram-negative bacteria as indicator bacteria, and the strains which can inhibit gram-positive bacteria and gram-negative bacteria and have the dual inhibition characteristic are selected; thirdly, fermentation characteristic test work; fourthly, inspection of influences of other factors on the strains; fifthly, inspection of influences of pH and temperature on bacteriocin crude extracts; sixthly, physiology and biochemistry identification work and molecule identification work.

Description

technical field [0001] The invention specifically relates to a method for isolation, screening and identification of Lactobacillus sake producing. Background technique [0002] Lactobacillus sake is a bacteriocin produced by lactic acid bacteria. Its polypeptide protein or protein complex with antibacterial activity is a natural food preservative and feed additive with potential development capabilities, which is of great significance to human life. Due to its wider and wider range of applications, its demand is also increasing day by day. For example, as a potential natural meat preservative, Lactobacillus sake can not only be used as a starter for fermented sausages, but can also impart good flavor and quality to sausages, and most of Lactobacillus sake is resistant to food-borne pathogens. Listeria monocytogenes has a strong inhibitory effect, and Lactobacillus sake has many types and different properties. However, its use and promotion are seriously affected due to its ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02C12Q1/04C12Q1/68C12R1/225
CPCC12N1/20C12N1/02C12Q1/04C12Q1/689
Inventor 刘学军王丽岩赵冬兵苟梦星于晓霞任峥峰王利媛
Owner JILIN AGRICULTURAL UNIV
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