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Lactobacillus plantarum Lp90, and screening method and application thereof

A technology of Lactobacillus plantarum and a screening method, applied in the field of Lactobacillus plantarum, can solve the problems of large individual differences in animal models, large human and material resources, large environmental differences, etc., and achieve the effects of low cost, obesity prevention, and high accuracy.

Active Publication Date: 2015-09-23
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although in vitro model screening is convenient and quick, it is quite different from the in vivo environment, while animal models have large individual differences, are expensive, and consume a lot of manpower and material resources

Method used

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  • Lactobacillus plantarum Lp90, and screening method and application thereof
  • Lactobacillus plantarum Lp90, and screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: Cultivation and isolation of Lactobacillus plantarum to be screened

[0032] Lactobacillus plantarum isolates naturally fermented pickle juice samples, takes samples and dilutes them with a sterile pipette, and adds them to 2% calcium carbonate and MRS medium at about 55°C, cultures at 37°C for 72 hours under anaerobic conditions, and selects the calcium-dissolving circle. Colonies with strong vitality were streaked and purified, and the purified strains continued to pass the primary screening: they did not grow at 45°C, grew at 15°C, and did not grow at pH 3.0. A total of 85 strains to be screened were obtained for cryopreservation.

[0033] Among them, MRS medium: maltose 5.0g, peptone 10.0g, beef extract 8.0g, yeast extract 4.0g, glucose 20.0g, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, magnesium sulfate 0.2g, manganese sulfate 0.05g, lemon Ammonium triammonium acid 2.0g, Tween 801.0g, agar 20.0g, calcium carbonate 2.0g, pH between 6.0 an...

Embodiment 2

[0034] Embodiment 2: Carry out strain screening and identification by nematode model:

[0035] The strains preserved at low temperature can be activated and expanded by the following steps: activate the primary screened strains through MRS liquid medium, and then inoculate the activated strains into a 10% (W / V) solids solution with a volume ratio of 1% to 3%. In milk, cultured at 43°C for 6 hours, and centrifuged to obtain the strain to be tested.

[0036] The nematode medium selected in this example is Op50-NGM medium. Cultured for 2 weeks (14 days).

[0037] Each 1000ml of Op50-NGM medium contains: 3g NaCl, 2.5g peptone, 17g agar, 1mol / L K 2 HPO 4 -KH 2 PO 4 (pH6.0) buffer solution 25ml, 1ml cholesterol solution, 1mol / L MgSO 4 1ml, 1mol / L CaCl 1ml.

[0038] For each strain to be selected, the nematode culture medium of the experimental group and the blank group with and without the strain to be screened were set at the same time. Wherein, the experimental group inclu...

Embodiment 3

[0049] Embodiment 3: control experiment

[0050] The commercially available Lactobacillus plantarum ST-III produced by Guangming Dairy has the functions of improving obesity, reducing body fat and reducing cholesterol.

[0051] The selected nematode medium is OP50-NGM medium, cultured for 2 weeks (14 days).

[0052] Set the nematode culture medium with and without adding Lactobacillus plantarum ST-III control group and blank group.

[0053] The concentration of Lactobacillus plantarum ST-III solution is 10 8 pieces / ml.

[0054] Fat staining: by Nile red and Sudan Black B staining. The brightness of staining after Nile red and the depth of staining after Sudan Black B staining were observed, and images of nematodes were taken with an Olympus microscope and analyzed with Image J software.

[0055] After Nile red staining, the comparison of staining intensity values ​​calculated by software scanning is shown in the table below:

[0056]

[0057] It is further ill...

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Abstract

The invention relates to lactobacillus plantarum Lp90, and a screening method and application thereof, in particular to lactobacillus plantarum with a function of inhibiting fat accumulation. The lactobacillus plantarum is named Lp90 and is preserved in the common microbiology center of the Chinese microbial strain preservation management committee, and the preservation number is CGMCC No.10453. The lactobacillus plantarum is obtained through an in-vitro nematode screening model, the fat accumulation of nematode can be reduced by 60 percent, and the lactobacillus plantarum has a potential function of preventing obesity and can be added into food, health care products or medicines as probiotics. Meanwhile, the invention provides the screening method namely a nematode culture model, and fat accumulation data of the nematode can be qualitatively and quantitatively detected, so that strains with the function of inhibiting the fat accumulation can be conveniently and quickly screened, the accuracy rate is high, and the economic cost is lower.

Description

technical field [0001] The invention relates to a plant lactobacillus and its screening method and application, in particular to a plant lactobacillus with the effect of inhibiting nematode fat accumulation. belongs to the field of microbiology. Background technique [0002] With the development of my country's national economy, people's living standards have generally improved, and more and more people are in a "sub-healthy" state and suffering from "civilized diseases". These diseases are difficult to be cured by drugs. Only by establishing a healthy lifestyle, especially following a healthy diet, can the prevention and alleviation be achieved. Therefore, it is necessary to regulate the balance of the internal environment and the circadian rhythm of the human body by supplementing foods rich in probiotics in a targeted manner. [0003] Probiotics are a type of bacteria that can promote human health. Common probiotics include Lactobacillus, Bifidobacterium and Streptococc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12Q1/04A23L1/30C12R1/25
Inventor 马凯方曙光姜甜冯胜鹏
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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