Induction of VBNC state of lactobacillus casei Zhang and detection method of cell fatty acid in VBNC state

A technology of Lactobacillus casei and cells, which is applied in the detection of fatty acids in VBNC cells and in the field of VBNC induction, can solve the problems of single induction method and few types of induction liquid, and achieve the effect of simple operation and strong stress resistance

Pending Publication Date: 2021-01-12
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

The existing probiotic Lactobacillus casei Zhang has a single induction method for the VBNC state, and there are few types of induction solutions that can be successfully induced, and there is no report on the mechanism of its cell membrane change

Method used

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  • Induction of VBNC state of lactobacillus casei Zhang and detection method of cell fatty acid in VBNC state
  • Induction of VBNC state of lactobacillus casei Zhang and detection method of cell fatty acid in VBNC state
  • Induction of VBNC state of lactobacillus casei Zhang and detection method of cell fatty acid in VBNC state

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1. Activation of Lb.casei Zhang

[0049] Lactobacillus casei Zhang (Lb.casei Zhang, CGMCC No.1697) preserved in vacuum freeze-drying was inoculated in liquid MRS medium, and activated for 18-24 hours at a temperature of 37°C. At the end of the logarithmic phase, activated lactobacilli were obtained. The said lactobacillus is Lactobacillus casei Zhang (Lb. casei Zhang), and the activation of the lactobacillus is carried out at a temperature of 37° C. for 18-24 hours.

[0050] Note: The probiotic Lb.casei Zhang was provided by the Lactic Acid Bacteria Culture Bank (LABCC), the Key Laboratory of "Dairy Biotechnology and Engineering" of the Ministry of Education, Inner Mongolia Agricultural University, LABCC number IMAU10048.

[0051] At the same time, it is also preserved in the China General Microbiological Culture Collection Center (China General Microbiological Culture Collection Center), and the preservation number is CGMCC No.1697.

[0052] 2. Inducing conditions

...

Embodiment 2

[0065] 1. Extraction of cell membrane fatty acids of Lb.casei Zhang

[0066] Obtain lactic acid bacteria sludge: regularly take the bacteria in the induction solution in Example 1 (in the process of normal state and VBNC state, VBNC state bacteria), centrifuge at 4000rpm for 5min, wash with normal saline once, weigh about 0.5g of bacteria in 10mL sterile, In a centrifuge tube with a screw cap.

[0067] Fatty acid extraction: Take 0.5g of the normal state and VBNC state cells of Lactobacillus casei Zhang, mix with 1.9mL methanol: chloroform (2:1), shake at room temperature for 15min, add 0.63mL chloroform and 0.63mL ultrapure water, make the chloroform and methanol 1:1, then shaken for 15 minutes, fully broken and centrifuged, so that the chloroform layer contains all lipids, while the methanol layer does not contain lipids, and the purified lipid extract is separated to obtain a chloroform layer.

[0068] B) The chloroform phase of the lower layer was removed, and the concent...

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Abstract

The invention provides an induction method of a VBNC state of lactobacillus casei Zhang and a detection method of cell membrane fatty acid in the VBNC state, and belongs to the technical field of microbial culture. The induction method comprises the following steps: inoculating the lactobacillus casei Zhang into a liquid MRS culture medium for activation, and then inoculating the lactobacillus casei Zhang into an MRS induction solution for induction in the VBNC state at 3.5-4.5 DEG C until the number of living cells counted by a plate colony counting method is 0 and the number of living cellscounted by a fluorescence microscope method is greater than 0 so as to obtain the VBNC state of the lactobacillus casei Zhang. By the method, cells in the VBNC state are successfully obtained, and thetypes of the VBNC state induced induction solution of the lactobacillus casei Zhang are enriched. The invention also provides a difference analysis result for extracting the components and contents of the membrane fatty acid in the normal state and the VBNC state of the lactobacillus casei Zhang.

Description

technical field [0001] The invention belongs to the technical field of microorganism culture, and in particular relates to a method for inducing the VBNC state of Lactobacillus casei Zhang and a method for detecting fatty acids in the VBNC state. Background technique [0002] "Probiotics" refer to live microorganisms that, when administered orally in sufficient quantities, improve the balance of the host's intestinal microbes, thereby having a beneficial effect on the health of the host. Lactic acid bacteria are the most representative genus of probiotics. Lactic acid bacteria and probiotics are not the same concept. Lactic acid bacteria refers to the general term for a class of bacteria that are Gram-positive, catalase-negative, and can use fermentable carbohydrates to produce more than 50% of lactic acid. There are many kinds of lactic acid bacteria. It has been found that lactic acid bacteria include 43 genera, 373 species and subspecies, and new lactic acid bacteria are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/00G01N30/02G01N30/06C12R1/245
CPCC12N1/20C12N1/005G01N30/02G01N30/06
Inventor 包秋华郭丽茹王俊国张猛代丽霞马学波
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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