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Lactobacillus plantarum producing non-protein small-molecule antibacterial metabolite and application of lactobacillus plantarum

A technology of Lactobacillus plantarum and metabolites, applied in the field of microorganisms, can solve problems such as inability to treat, drugs cannot meet the needs of treatment, high mortality, etc., and achieve the effect of facilitating stable performance and good development and application prospects.

Active Publication Date: 2019-11-15
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current status of antibacterial drugs is: on the one hand, existing drugs cannot meet the needs of treatment; on the other hand, there are not enough new antibacterial drugs developed to replace old or gradually ineffective drugs
As a result, many common infections gradually become dangerous or untreatable, leading to longer treatment cycles and higher mortality, as in the days before antibiotics

Method used

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  • Lactobacillus plantarum producing non-protein small-molecule antibacterial metabolite and application of lactobacillus plantarum
  • Lactobacillus plantarum producing non-protein small-molecule antibacterial metabolite and application of lactobacillus plantarum
  • Lactobacillus plantarum producing non-protein small-molecule antibacterial metabolite and application of lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Screening and identification of antibacterial lactic acid bacteria

[0032] (1) Isolation of microbial strains

[0033] The pickle water samples were diluted with sterilized saline in a 10-fold dilution gradient. Take 100μL of 10 -2 、10 -3 、10 -4 、10 -5 The diluted kimchi water dilution was spread on the MRS solid medium, and incubated at a constant temperature of 37° C. for 48 hours. A single colony of lactic acid bacteria was picked, and the characteristic colony was further isolated and purified repeatedly by the streak separation method, and the morphological characteristics of each colony were recorded and observed by Gram staining microscope.

[0034] The MRS solid medium is composed of the following components in terms of mass percentage: 2% glucose, 1% peptone, 1% beef extract, 0.5% yeast extract, 0.5% sodium acetate, 0.1% Tween 80, dihydrogen citrate 0.1% of amine, 0.02% of magnesium sulfate, 0.005% of manganese sulfate, 1.5% of agar powder,...

Embodiment 2

[0043] Embodiment 2: the antibacterial activity of plantarum lactobacillus KZ0310

[0044] Seven pathogenic bacteria, Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii and Pseudomonas aeruginosa, were activated. Each pathogenic bacterium of 0.1mL overnight culture is inoculated in 100mL fresh LB agar medium (0.7wt% agar), mixes, and uses the double-layer streaking method described in embodiment 1 (2) to measure Lactobacillus plantarum KZ0310 The diameter of the inhibition zone against these 7 pathogenic bacteria (calculated in the direction perpendicular to the bacterial line).

[0045] The result is as figure 2 As shown, Lactobacillus plantarum KZ0310 showed good antibacterial activity against seven pathogenic bacteria, and the order of antibacterial ability was: Listeria monocytogenes > Escherichia coli > Pseudomonas aeruginosa bacteria>Salmonella>Cronobacter sakazakii>Staphylococcus aureus>Enterococ...

Embodiment 3

[0046] Example 3: Extraction, partial purification and antibacterial activity of antibacterial metabolites

[0047] (1) Extraction of antibacterial metabolites

[0048] A single colony of Lactobacillus plantarum KZ0310 was picked and inoculated in 10 mL of MRS liquid medium, and cultivated overnight at 37°C and 150 rpm as the seed fungus. Inoculate the seed bacteria into 1L MRS liquid medium with an inoculation amount of 0.1wt%, and culture it statically at 30°C for 72h, centrifuge (8000rpm, 15min, 4°C), extract the supernatant with ethyl acetate, and collect the milky white liquid in the middle layer , Ethyl acetate was removed by rotary evaporation, and a crude sample of metabolites of Lactobacillus plantarum KZ0310 was obtained.

[0049] Using the Oxford cup method to determine the pathogenicity of Enterococcus faecalis, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Salmonella, Enterobacter sakazakii and Pseudomonas aeruginosa Antibacterial activity of ...

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Abstract

The invention discloses lactobacillus plantarum KZ0310 with a preservation number of CGMCC No.17712. The thallus and the metabolite thereof have broad-spectrum antibacterial activity, show strong bacteriostatic capability to various Gram-positive bacteria and Gram-negative bacteria (including pathogenic bacteria and putrefying bacteria), can be used for preparing antibacterial or / and preservativeproducts, and can be applied to the fields of medical treatment, food and the like to control bacterial pathogenic bacteria or putrefying bacteria. The antibacterial metabolite is different from lactobacillus bacteriocin or antibacterial peptide, is a non-protein small molecular substance, and can be prevented from being degraded and inactivated by protease in human, animal and plant bodies when in use, so that the antibacterial metabolite is beneficial to stable development of antibacterial activity, and has good development and application prospects.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and relates to a plant lactobacillus producing non-protein small molecule antibacterial metabolites and an application thereof. Background technique [0002] Antimicrobials, including antibiotics, have not only saved countless lives, but have also helped people live longer and improve their quality of life. Unfortunately, however, the overuse and misuse of antimicrobials (including inappropriate use in animals, food, agriculture, and aquaculture for non-therapeutic purposes) has led to the emergence and massive spread of resistant microorganisms. Antimicrobial resistance is found in all microorganisms, whether bacteria, fungi, viruses or parasites. With the rapid development of global trade and travel, resistant microorganisms can spread to any corner of the world. However, the current status of antibacterial drugs is: on the one hand, existing drugs cannot meet the needs of treatment; o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P1/04A01N63/00A01P1/00C12R1/25
CPCA01N63/00C12P1/04C12N1/205C12R2001/25
Inventor 易兰花曾凯芳邓丽莉祁腾阮长晴罗惟
Owner SOUTHWEST UNIVERSITY
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