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Lactobacillus plantarum ZJ316, procreant antibiotic peptides, preparation and use thereof

A ZJ316, antimicrobial peptide technology, applied in Lactobacillus plantarum ZJ316, the antimicrobial peptide produced and its preparation and application field, can solve the problems of basic research knowledge to be accumulated, lactic acid bacteria antibacterial peptide research started late, etc., and achieve the effect of high-efficiency bactericidal ability

Active Publication Date: 2009-01-28
ZHEJIANG GONGSHANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Our country is very rich in lactic acid bacteria resources, but the research on lactic acid bacteria antimicrobial peptides started relatively late, and the corresponding basic research knowledge has yet to be accumulated

Method used

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  • Lactobacillus plantarum ZJ316, procreant antibiotic peptides, preparation and use thereof
  • Lactobacillus plantarum ZJ316, procreant antibiotic peptides, preparation and use thereof
  • Lactobacillus plantarum ZJ316, procreant antibiotic peptides, preparation and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Isolation of Lactobacillus plantarum ZJ316

[0038] Use sterile tweezers to pick up a little baby feces (provided by Children's Hospital of Zhejiang University) and culture it in MRS liquid medium (prepared according to the following formula: 10 g of peptone, 10 g of beef extract, 6 g of yeast extract, 2 g of dipotassium hydrogen phosphate, tricitric acid amine 2g, sodium acetate 5g, glucose 20g, magnesium sulfate heptahydrate 0.2g, manganese sulfate tetrahydrate 0.05g, Tween 1mL, pH6.3, deionized water to make up to 1000mL), enrich the bacteria in a constant temperature incubator at 37°C Incubate for 24 hours. Then take one drop of the enrichment solution and put it on the plate of MRS solid medium (take the aforementioned 1000mL MRS liquid medium and add 18g agar), spread it evenly, and cultivate it in an anaerobic incubator at 37°C for 48 hours. The samples were taken for marking on the plate, cultured in an anaerobic incubator at 37°C for 24 hours, and s...

Embodiment 2

[0066] Example 2: Lactobacillus plantarum ZJ316 fermentation preparation of antimicrobial peptide plantaricin ZJ316MRS liquid medium preparation: peptone 10g, beef extract 10g, yeast extract 6g, dipotassium hydrogen phosphate 2g, triamine citrate 2g, sodium acetate 5g, glucose 20g, seven Magnesium sulfate water 0.2g, manganese sulfate tetrahydrate 0.05g, Tween 1mL, the above mixture was dissolved in 960mL deionized water, adjusted to pH 6.4 with 1mol / L NaOH, added deionized water to a total volume of 1L, and steam sterilized under high pressure 20min.

[0067] Preparation of MRS solid medium: Take 1 liter of the above-prepared MRS liquid medium, add 15 g of agar powder under heating and stirring, and condense to obtain MRS solid medium.

[0068] Culture conditions: The preserved Lactobacillus plantarum ZJ316 strain was taken out from -20°C, streaked on MRS solid medium, and cultured anaerobically at 37°C for 24 hours. Single colonies were inoculated in MRS liquid medium, cult...

Embodiment 3

[0069] Example 3: Isolation and functional identification of the antimicrobial peptide plantaricin ZJ316 in Lactobacillus plantarum ZJ316

[0070] 1. Tolerance to pH

[0071] Use 1mol / L of HCl and 1mol / L of NaOH to adjust the pH of the fermentation supernatant to: 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, do Oxford cup experiments, and use Micrococcus luteus As an indicator bacterium, 1mol / L HCl was used as a control to observe the change in the size of the inhibition zone. After testing, the control 1mol / L HCl has no inhibitory effect on the indicator bacteria. This proves that it is the plant lactobacillus plantaricion ZJ316 that has antibacterial effect, and it has antibacterial activity in an acidic environment, and it is inactivated under the condition of pH>4; and the higher the acidity, the stronger the activity ( figure 2 ).

[0072] 2. Stability to temperature

[0073] Treat the fermentation supernatant at -20°C, 4°C, 37°C, 60°C, 90°C, 100°C, and 121°C fo...

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Abstract

The invention provides a lactobacillus plantarum ZJ316 which is separated from excrement of infants and anti-bacterial peptide which is produced by the fermentation of the lactobacillus plantarum ZJ316, namely plantaricin ZJ316, and preparation and application thereof; the plantaricin ZJ316 is a novel lactobacillus anti-bacterial peptide with high efficiency and small molecular; the anti-bacterial peptide has bacteriostatic action of broad spectrum; due to small molecular weight, a lipid bilayer is hard to be crossed so as to form a complex body with small holes to be different from the action mode of nisin; but the anti-bacterial peptide also has sterilizing capacity with high efficiency, is a lantibiotic anti-bacterial peptide with superhigh application and development potential and can be applied to preparing the alternative medicine of feed additives, food preservatives and antibiotic, etc.

Description

(1) Technical field [0001] The invention relates to a new strain of Lactobacillus plantarum-Lactobacillus plantarum ZJ316, and an antimicrobial peptide produced by fermentation thereof-plantaricin ZJ316 (plantaricin ZJ316) and its preparation and application. (2) Background technology [0002] Alexander Fleming's discovery of penicillin in 1928 saved thousands of lives. However, the problem of antibiotic resistance has become increasingly serious in recent years, bringing new dangers to people's lives and health. Research and development of new antibiotics has become very urgent. Researchers are working hard to find suitable antimicrobial peptides to replace antibiotics. Lactic acid bacteria are food-grade micro-organisms (GRAS) recognized by the FDA as safe, and lactic acid bacteria antimicrobial peptides will be researched and developed as natural and safe food preservatives and pharmaceutical products. Lactic acid bacteria antimicrobial peptides have been favored by re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C07K14/195C12P21/02C07K1/14A23K1/16C12R1/25A23K20/147
Inventor 顾青
Owner ZHEJIANG GONGSHANG UNIVERSITY
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