Screening method and application of lactobacillus plantarum

A technology of Lactobacillus plantarum and Lactobacillus, applied in application, microorganism-based methods, microorganisms, etc., can solve problems such as adverse effects, reduced nutritional value, infection of spoilage bacteria, etc., and achieve the effect of good thermal stability

Inactive Publication Date: 2015-09-02
TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Adding chemical preservatives is an important food preservation method used by humans, but long-term studies have found that excessive consumption of synthetic preservatives has adverse effects on blood pressure, heart, kidneys, etc., and even some chemical preservatives are carcinogenic, teratogenic and easy to cause food poisoning etc.
Thermal sterilization is also an important method to kill microorganisms in the process of food processing, but for some low-temperature products, if the heating temperature is too high, the color and flavor will change significantly, and the nutritional value will also be greatly reduced
Other physical methods: such as ultraviolet disinfection, filter sterilization, cobalt 60 irradiation, etc., have no toxic effect on the human body, but when the package is opened, there is a possibility of re-infection with spoilage bacteria

Method used

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  • Screening method and application of lactobacillus plantarum
  • Screening method and application of lactobacillus plantarum
  • Screening method and application of lactobacillus plantarum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: the acquisition of Lactobacillus plantarum.

[0020] One: the acquisition of wild Lactobacillus plantarum.

[0021] Source of bacteria: isolated from homemade yogurt. Under aseptic conditions, take 5 mL of yogurt, add it to 45 mL of sterile distilled water, shake fully for 5 minutes, and serially dilute by 10 times, starting from 10 -1 ~ 10 -7 Gradient, choose the appropriate gradient, take 100uL and apply it on the 3 MRS (peptone 10g, yeast extract 5g, beef extract 10g, glucose 20g, dipotassium hydrogen phosphate 2g, ammonium citrate 2g, sodium acetate 5g, magnesium sulfate 0.58g, manganese sulfate 0.25g, Tween 80 1mL, agar 15g, Distilled water 1L, pH6.50) culture medium, 30 ℃ static culture and count. Select a single colony with obvious calcium dissolution circle, carry out Gram staining test, and microscopic examination. Gram-positive bacteria were selected for streak isolation and purification for 2 to 3 times, and the isolated and purified sing...

Embodiment 2

[0063] Example 2, identification of antibacterial effect of Lactobacillus plantarum) CGMCC No 5297.

[0064] From Example 1, it can be seen that plant lactobacillus (CLactobacillus plantarum) CGMCC No 5297 of the present invention has a strong inhibitory effect on Listeria monocytogenes, so it is speculated that this bacterial strain can secrete bacteriocin, and the plant lactobacillus (CLactobacillus plantarum) can be treated by the following method The antibacterial effect of CGMCC No 5297 and its secreted bacteriocin were identified.

[0065] 1. Eliminate factor interference.

[0066] 1. Eliminate acid interference.

[0067] The test method for inhibiting pathogenic bacteria is the Oxford cup double-layer plate method: LB culture medium for indicator bacteria containing 1.5% agar: 10 g of egg white, 5 g of yeast extract, 10 g of sodium chloride, 1.0 L of steamed water, pH7. 0. Pour 10mL per plate into a sterile Petri dish and cool it in a clean bench. Gently place the ...

Embodiment 3

[0094] Example 3 Inhibition of Lactobacillus plantarum CGMCC No 5297 on Listeria monocytogenes.

[0095] 1. Effect of bacteriocin produced by Lactobacillus plantarum CGMCC No 5297 on the growth curve of Listeria monocytogenes.

[0096] Use an inoculation loop to pick a single colony of Listeria monocytogenes CVCC1595 in 5ml LB medium, and culture overnight at 37°C. Inoculate in fresh LB medium according to 1% inoculum amount, and culture at 37°C until plateau stage. Dilute with fresh LB to about 10 bacteria 6 cfu. Then each test tube was divided into 2.75ml and divided into 4 groups. Group 1 added 250ul MRS medium as the control. Group 2 added 250ul CGMCC5297 sterile supernatant, Group 3 added 125ul CGMCC5297 sterile supernatant + 125ul MRS medium, Group 4 added 62.5ul CGMCC5297 sterile supernatant + 187.5ul MRS medium. The pH of the sterile supernatant of the above CGMCC5297 was adjusted to 6.0. Put 4 groups of test tubes together at 30°C for static culture, take one tu...

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Abstract

The invention discloses a mutation breeding method of lactobacillus plantarum and application of a strain of the lactobacillus plantarum. The collection number of the lactobacillus plantarum in China general microbiological culture collection center is CGMCC No 5297. The lactobacillus plantarum can generate bacteriocin which has a better inhibiting effect on multiple gram-positive bacteria and especially has great a high inhibiting effect on listeria monocytogenes, and the bacteriocin has the advantages of better heat stability, acid stability and the like and can be applied as a food additive and a feed preservative agent which have wide application prospect.

Description

technical field [0001] The invention relates to a plant lactobacillus and its application. Background technique [0002] Food safety is an important issue worldwide, especially in developing countries like my country. In recent years, the number of patients with foodborne diseases has remained high, and food poisoning caused by microorganisms is very serious. Adding chemical preservatives is an important food preservation method used by humans, but long-term studies have found that excessive consumption of synthetic preservatives has adverse effects on blood pressure, heart, kidneys, etc., and even some chemical preservatives are carcinogenic, teratogenic and easy to cause food poisoning etc. Thermal sterilization is also an important method to kill microorganisms during food processing, but for some low-temperature products, if the heating temperature is too high, the color and flavor will change significantly, and the nutritional value will also be greatly reduced. Other...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12P21/00A23L3/3571A23K1/16C12R1/25
Inventor 郑雯宋诙马延和谭明徐健勇
Owner TIANJIN INST OF IND BIOTECH CHINESE ACADEMY OF SCI
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