Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin

A technology of Lactobacillus plantarum and bacteriocin, applied in the field of microbial application, can solve the problems of poor pertinence, difficulty in obtaining strains that meet production requirements, and large workload.

Active Publication Date: 2011-05-04
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, finding strains with excellent performance from nature has become a very important task. When using conventional methods to isolate strains from nature, th

Method used

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  • Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin
  • Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin
  • Lactobacillus plantarum, method for fermenting and preparing bacteriocin of Lactobacillus plantarum, and application of Lactobacillus plantarum and bacteriocin

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Screening and identification of strains of embodiment 1

[0047] 1. Isolation of Lactobacillus in Habitat

[0048] In the initial screening, the collected samples are first diluted with physiological saline, generally 3 to 4 orders of magnitude (depending on the sample, the bacteria in the sample can be diluted to 5 orders of magnitude, and the rare content can be diluted to 2 orders of magnitude), Add 100ul of the last two dilutions of the bacteria-containing solution to the modified MRS plate containing 0.02% bromocresol purple indicator, spread evenly, and select on the MRS medium for 24 hours at 37 °C to change the bromocresol purple from purple. The yellow colonies were isolated and cultured, and the obtained pure bacteria were marked and numbered respectively.

[0049] 2. Preliminary screening of antibacterial by seeding method

[0050] Take pathogenic Escherichia coli CVCC195 and Salmonella pullorum CVCC79301 as indicator bacteria, take their seed bacteria liqu...

Embodiment 2

[0058] Example 2 Physiological and biochemical properties of Lactobacillus plantarum strain YJG

[0059] 1 titer standard curve and fitting equation

[0060] After the fermentation of strain YJG, the centrifuged supernatant still had bacteriostatic activity at a dilution of 1 / 128, but no bacteriostatic activity at a dilution of 1 / 256, and the bacteriostatic activity of YJG was 1280AU. / mL.

[0061] The standard curve was made by measuring the potency value by one-dose method, and the titer fitting equation of strain YJG was obtained by SAS statistical analysis of the measured data:

[0062] y=2.8540+0.0862xR 2 =0.9392

[0063] x- The diameter difference of the inhibition zone between the sample to be tested and the central concentration bacteriocin solution

[0064] y - the value of the sample to be tested is the value obtained by taking log10

[0065] R 2 - Correlation coefficient

[0066] 2. Growth curve and titer change curve

[0067] The growth curve and antibacter...

Embodiment 3

[0068] The preparation of embodiment 3 lactobacillus and its bacteriostatic effect

[0069] 1. Preparation of Lactobacillus

[0070] 1. Preparation of Seed Liquid

[0071] The YJG was inserted into the sterilized liquid medium and cultivated at 30°C for 36 hours to obtain the seed liquid, and the number of viable bacteria was about 10. 7 ~10 9 cfu / ml.

[0072] The liquid culture medium is prepared according to the following proportions: 10 g of peptone, 10 g of beef extract, 5 g of yeast extract, 18 g of glucose, 2 g of ammonium citrate, 4 g of sodium acetate trihydrate, 2 g of dipotassium hydrogen phosphate, 801 mL of Tween, and 0.2 g of magnesium sulfate. g, manganese sulfate 0.04, water 1000mL.

[0073] 2. Large-scale fermentation preparation of Lactobacillus plantarum fermentation broth

[0074] Take 5kg of soybean meal, 3kg of corn gluten, 10kg of brown sugar, and 10kg of molasses into the fermentation tank, add 280kg of water and mix, sterilize at 0.1-0.13MPa, 115-1...

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Abstract

The invention provides a new Lactobacillus plantarum bacterial strain YJG which was preserved in the China General Microbiological Culture Collection Center (CGNCC) with the collection number of CGMCC No. 2994 on march, 30th, 2009. The Lactobacillus plantarum bacterial strain YJG can produce broad-spectrum bacteriostatic activity bacteriocin with high yield, and can express bacteriostasis for staphylococcus aureus, salmonella and E.coli of gram-negative bacterium, L. plantarum, L.brevis, Streptococcus and Bacillus of gram-positive bacterium, and the like. A virus attacking tests with high dosage proves that the Lactobacillus plantarum bacterial strain YJG is safe to animal bodies. The Lactobacillus plantarum bacterial strain YJG and a zymophyte liquid thereof can be used for preparing additives of green animal feeds, and the bacteriocin of the Lactobacillus plantarum bacterial strain YJG can be used for preparing green biological veterinary drugs, environmental-friendly biopreservatives, biocontrol drugs and the like.

Description

technical field [0001] The invention relates to the application technology of microorganisms, in particular to a new strain of Lactobacillus plantarum with high production of broad-spectrum bacteriocin, and the invention also relates to the application of the bacterial strain and the bacteriocin it produces. Background technique [0002] Since the 1950s, antibiotics, chemical synthetic drugs and other growth-promoting health care agents have been widely used for a long time and in large quantities in animal feeding and feed, resulting in serious drug residues in livestock and aquatic products, deterioration in the quality of livestock and poultry products, and multiple bacteria. The increase of drug-resistant and drug-resistant strains poses a great threat to the health and safety of human beings and the ecological environment. Due to the many side effects of antibiotics, especially the concerns of consumers on food safety in recent years, countries around the world are bann...

Claims

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Application Information

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IPC IPC(8): C12N1/20C07K14/335C07K1/36C07K1/30C07K1/20C07K1/16C12P21/00A61K35/74A61K38/16A61P31/04A23K1/16A23K1/17C12R1/25A23K10/18A23K20/147A61K35/747
Inventor 张日俊刘开健韩冰黄燕
Owner CHINA AGRI UNIV
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