Lactobacillus plantarum GL-5 with oxidation resisting activity and application thereof

A Lactobacillus plantarum, antioxidant activity technology, applied in the field of microorganisms, can solve the problems of gastrointestinal intolerance, weak bile salt tolerance, weak antioxidant activity, etc., and achieve good gastrointestinal tolerance and bile salt tolerance. Tolerance, effect of strong antioxidant activity

Active Publication Date: 2020-06-19
LANZHOU UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the antioxidant activity of LGG bacteria is weak
[0005] Although a variety of Lactobacillus plantarum with antioxidant activity have been screened out, there are generally disadvantages such as poor antibacterial effect or gastrointestinal intolerance.
Among them, Chinese patent CN109182162A provides a plateau Lactobacillus plantarum, which has a certain antioxidant activity and can effectively scavenge free radicals in the body, but its survival rate in intestinal juice is low and its gastrointestinal tolerance is weak; Chinese patent CN104789511A provides a strain of Lactobacillus plantarum AB-2, which has broad-spectrum antibacterial properties, but has weak tolerance to bile salts

Method used

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  • Lactobacillus plantarum GL-5 with oxidation resisting activity and application thereof
  • Lactobacillus plantarum GL-5 with oxidation resisting activity and application thereof
  • Lactobacillus plantarum GL-5 with oxidation resisting activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Isolation and activity detection of Lactobacillus plantarum

[0024] 1.1 Separation

[0025] Take about 0.5g of fermented dairy products made by herdsmen in the Guoluo area of ​​Qinghai at an altitude of 4000m above sea level, and dilute them to 10 with PBS. -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 , 10 -6 , 10 -7 , 10 -8 The concentration gradient of each concentration is 100μL and spread evenly on the modified MRS solid medium with calcium carbonate. Each concentration is repeated twice. Place it in a constant temperature incubator at 37°C for 48 hours and observe the colony and transparent circles to facilitate further separation and purification.

[0026] 1.2 Purification culture

[0027] Choose a suitable gradient from the above cultured concentration gradients, select 15-25 single colonies with clear borders and transparent circles, and the selected colonies are subcultured on the newly prepared MRS solid medium by streaking for 24- For 48h, streak culture again, thi...

Embodiment 2

[0033] Example 2 Identification and preservation of strain GL-5

[0034] 2.1 Sequence analysis of 16S rRNA of strain GL-5

[0035] Extract the DNA of strain GL-5, use bacterial 16S rRNA universal primers for PCR amplification:

[0036] The upstream primer is 27F AGAGTTTGATCMTGGCTCAG

[0037] Downstream primer is; 1492R GGTTACCTTGTTACGACTT

[0038] PCR amplification system;

[0039] 50μL total reaction system includes: 2×Mix Taq, 25μL; Primers (10μM): Upstream primer (Forward), 2.0μL, Downstream primer (Reverse), 2.0μL; DNA template (100ng / μL), 2.0μL; ddH2O, 19μL .

[0040] PCR reaction conditions:

[0041] Pre-denaturation: 94°C, 4min; 94°C denaturation, 1min; 55°C renaturation, 1min; 72°C extension, 1.5min, 30 cycles in total; 72°C extension for 10min.

[0042] The reaction was performed on a Bio-Rad iCycler Thermal Cycler (Bio-Rad Laboratories, Hercules, CA) instrument. The amplified products were sent to Shanghai Shenggong Biotechnology Co., Ltd. for sequencing. The sequencing results...

Embodiment 3

[0046] Example 3 Tolerance of strain GL-5 to simulated gastric juice and intestinal juice

[0047] Gastric juice preparation: prepare a PBS buffer with a pH of 7.4, add an appropriate amount of dilute hydrochloric acid to adjust the pH to 3.0, and dilute to 100ml, then weigh 1.0g of pepsin and add it to it, filter with a 0.22μm filter, and set aside.

[0048] Intestinal juice preparation: prepare PBS buffer with pH value of 8.0, dilute to 100ml, weigh 0.3g porcine bile salt, 1.0g trypsin, mix into it, filter with 0.22μm filter, and set aside.

[0049] Bacterial solution configuration: Take the strain GL-5 stored in 50% glycerol, inoculate 1.5ml of MRS liquid medium according to the inoculum amount of 2%, and place it in a constant temperature incubator at 37°C for 10 hours as a seed bacterial solution for the next experiment of. The seed bacteria liquid was inoculated into 5ml MRS liquid medium at an inoculum of 2% and placed in a 37°C constant temperature incubator for 18 hours for...

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Abstract

The invention belongs to the technical field of microbes and particularly relates to lactobacillus plantarum GL-5 with oxidation resisting activity and application thereof. The lactobacillus plantarumGL-5 with oxidation resisting activity disclosed by the invention is screened out from a home-made fermented dairy product of a herdsman's house of a Qinghai Guoluo region with an altitude of 4,000mor more, is collected in China General Microbiological Culture Collection Center of China Committee for Culture Collection of Microorganisms on November, 21, 2019 and has an accession number of CGMCCNo. 18988. The lactobacillus plantarum GL-5 has a 16S rRNA sequence shown in SEQ ID NO: 1. The lactobacillus plantarum GL-5 provided by the invention has relatively good bacteriostatic activity, and activity of enteropathogenic bacteria, i.e., staphylococcus aureus and escherichia coli can be remarkably inhibited; the lactobacillus plantarum GL-5 provided by the invention has relatively high oxidation resisting activity and can remarkably tolerate catalase; and the lactobacillus plantarum GL-5 provided by the invention has relatively high gastrointestinal fluid tolerance and cholate tolerance,and the survival rate of the lactobacillus plantarum GL-5 after culture in simulated gastric fluid and intestinal fluid is remarkably higher than that of lactobacillus rhamnosus LGG.

Description

Technical field [0001] The invention belongs to the technical field of microorganisms, and specifically relates to a strain of Lactobacillus plantarum GL-5 with antioxidant activity and its application. Background technique [0002] Oxidation is a necessary metabolic process of the body, but excessive oxidation can cause damage to biological macromolecules, leading to the occurrence of related diseases such as aging of the body. Free radicals have a strong oxidative effect, which can trigger lipid peroxidation and cause cell membrane damage. Damage, which in turn causes cell death. When the human body produces excessive free radicals, the free radicals will combine with biological macromolecules such as proteins and nucleic acids in human cells and destroy their structure and function. The main hazards of free radicals to the human body include: reducing resistance, weakening cell renewal rate, causing oxidative fatigue, leading to inflammation and aging. [0003] Probiotics are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L33/135A61K35/747A61P31/04A61P39/06A61P39/00C12R1/25
CPCC12N1/02A23L33/135A61K35/747A61P31/04A61P39/06A61P39/00A23V2002/00C12R2001/25C12N1/205A23V2400/169A23V2200/3204A23V2200/30Y02A50/30
Inventor 张春江扎拉加李玉玺白妍睿刘冠兰
Owner LANZHOU UNIVERSITY
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