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Novel lactobacillus plantarum and use thereof

a technology of lactobacillus and plantarum, applied in the field of microorganisms, can solve the problems of affecting the color and flavor of pickles, reducing the number of viable bacteria, and human beings and animals are difficult to survive healthily, and achieves strong genetic stability and rapid degradation of nitrites

Inactive Publication Date: 2017-01-12
NINGXIA RISINGMARK INTPROP CONSULTING CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014]It is found through tests that the Lactobacillus plantarum tlj-2014 provided in the present invention is strongly genetically stable, since it can be continuously slant-subcultured for 10 generations without obvious change in traits and with various performance criteria kept normal. With the strain, the lactic acid production rate can be up to 35 g / L / d, and the concentration of lactic acid after 71 hrs of fermentation is up to 95 g / L. The Lactobacillus plantarum tlj-2014 which can survive at pH 1.80, is tolerant to 1% bile salt, and can degrade the nitrite quickly with a decomposition capability of up to 9.8 mg / h / kg. When the strain is used in production of pickles, the nitrite concentration is lower than 5 mg / kg throughout the entire fermentation process, which is far below the content (20 mg / kg) specified in the national standard GB2714-2003.

Problems solved by technology

It is believed that once cessation of growth of the lactic acid bacteria occurs, human beings and animals are hard to survive healthily.
However, if the tolerance of the start culture for artificial inoculation to the environment is insufficient, the color and flavor of the pickles and especially the number of the viable bacteria are affected.
In the patent, the strains used for fermentation are Lactobacillus plantarum LP-115, Leuconostoc mesenteroides LM-57, and Pediococcus acidilactici P-751, which have a low tolerance to the environment, and tend to suffer from decreased number of viable bacteria during the storage and selling of the pickles, so that the healthcare functions of the viable bacteria cannot be highly exerted.
During the fermentation of the pickles, a safety problem that is difficult to overcome exists, that is, the production of the nitrite.
The nitrite is very harmful to human body, poisoning may occur upon uptake in an amount of 0.2-0.5 g, and the nitrite is also a precursor of the highly carcinogenic substance nitrosamine.
However, the salt content in the pepper is too high, and long-term consumption of the pepper may cause damage to human body.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

Characteristics of the Strain

[0015]The Lactobacillus plantarum tlj-2014 is deposited on Jul. 2, 2014 in China General Microbiological Culture Collection Center (CGMCC) (Institute of Microbiology, Chinese Academy of Sciences, Building 3, No. 1 West Beichen Road, Chaoyang District, Beijing, China, 100101) under CGMCC Accession No. 9405.

[0016]The strain has the following features. When observed under a microscope, the strain is short rod-like and gram positive, has no flagellum, and is asporulate. When cultured on a solid medium, the Lactobacillus plantarum tlj-2014 forms a white colony, which has a smooth surface, and is dense and in a circular morphology with a regular edge.

[0017]The physiochemical properties include hydrogen peroxidase (−), gelatine liquefication (−), indole test (+), mobility (−), gas from fermentation (−), nitrite reduction (−), gas from fermentation (−), hydrogen sulfide production (−), and growth in MRS medium pH 4.0 (+).

example 2

Screening of the Strain

[0018]The Lactobacillus plantarum tlj-2014 according to the present invention was screened from the original starting strain Lactobacillus plantarum L after mutagenesis, and the original starting strain Lactobacillus plantarum L was collected by Li Zheng from the ensilage in a sheep breezing farm in Yanchi, Ningxia on Sep. 15, 2013.

[0019]When grown in a MRS medium with glucose, the starting strain had a lactic acid production rate of 1.5 g / L / d, almost stopped growing when the culture medium reached pH 3.5, and had a decomposition rate for sodium nitrite of 0.34 mg / h / kg of Chinese cabbage.

[0020]In order to improve the lactic acid production rate, the acid tolerance, and the nitrite decomposition rate, the strain was subjected to DES and NTG mutagenesis sequentially. After mutagenesis, the strains were primarily screened in a plate containing MRS supplemented with calcium carbonate, and then the high production strains were secondarily screened by fermentation i...

example 3

Tests in 5 L Fermentor

[0039](1) The Lactobacillus plantarum L2 was picked up by using an inoculating loop from the test-tube slant, inoculated in a 250 mL conical flask containing 50 mL MRS medium (agar free, and containing 150 g / L of glucose), and incubated at 200 rpm and 37° C. for about 12 hrs, such that the bacteria were in a middle exponential growth phase.

[0040](2) The strain in the exponential phase was inoculated in a 5 L fermentor containing 3 L of a liquid MRS medium (with glucose in an initial concentration of 150 g / L) in an amount of 10%, and incubated at 37° C. and 100 rpm for 8 hrs. The dissolved oxygen was controlled to be 10% (through aeration at a rate of 0.5 L / min) in the earlier exponential phase, and then anaerobic incubation was continued for 63 hrs in the later phase. After fermentation, the Lactobacillus plantarum L2 was shown to have a lactic acid production of 95 g / L. Such a lactic acid production rate can promote the rapid fermentation of pickles.

[0041](3) ...

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Abstract

A novel Lactobacillus plantarum and use thereof are disclosed, which fall within the technical field of microorganisms. The Lactobacillus plantarum tlj-2014 is deposited in China General Microbiological Culture Collection Center (CGMCC) under CGMCC Accession No. 9405. With the strain, the lactic acid production rate can be up to 35 g / L / d, and the concentration of lactic acid after 71 hrs of fermentation is up to 95 g / L. The Lactobacillus plantarum is acid tolerant, survives well at pH 1.80, can degrade the nitrite quickly with a decomposition capability of up to 9.8 mg / h / kg, and is tolerant to 1% bile salt. When the strain is used in production of pickles, the nitrite concentration is less than 5 mg / kg throughout the entire fermentation process, which is far below the content specified in the national standard GB2714-2003.

Description

CROSS-REFERENCE TO RELATED APPLICATION[0001]This application claims the priority of Chinese Patent No. 201410478590.9 filed on Sep. 18, 2014 and entitled “Novel Lactobacillus plantarum and Use Thereof”.TECHNICAL FIELD[0002]The present invention falls within the technical field of microorganisms, and more specifically relates to a novel Lactobacillus plantarum and use thereof.BACKGROUND[0003]Lactic acid bacteria (LAB) are a generic term of a class of bacteria with which large quantities of lactic acid can be produced from fermentable carbohydrates. This class of bacteria is widely distributed in nature, is abundant in species diversity, and has great application value in many important areas intimately related to human life including industry, agriculture and animal husbandry, food, medicines, and others. The use of lactic acid bacteria in food can increase the nutritive value, improve the flavor, and enhance the preservation and added value of the food. Also numerous studies suggest...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A23B7/155C12R1/25C12N1/20A23B7/10
CPCA23B7/155A23B7/10A23Y2220/67C12N1/20A23V2002/00C12R1/25C12R2001/25C12N1/205A23V2400/169
Inventor LI, ZHENG
Owner NINGXIA RISINGMARK INTPROP CONSULTING CO LTD
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