Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof

A technology of antibacterial activity and lactic acid bacteria, applied in the field of microorganisms, can solve problems such as intolerance to bile salts and failure to grow, and achieve good antibacterial effect

Inactive Publication Date: 2015-05-13
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, as probiotics, currently reported Lactobacillus salivarius has disadvantages such as inability to grow at low pH, or intolerance to bile salts.

Method used

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  • Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof
  • Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof
  • Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Screening, domestication and isolation of Lactobacillus salivarius (L.salivarius) GZPH2

[0036] Randomly buy kimchi samples from the Guangzhou market, take 1mL of sample liquid and add 50mL of aseptically treated MRS liquid (adding 0.05% CaCO 3 ), 150r / min 37 ℃ enrichment culture for 12h; the enrichment solution was diluted to 10 with sterile physiological saline -8 , spread each dilution gradient on the plate in turn, culture at 37°C for 24 hours, select a plate with a single colony number of 30-300, and select a single colony with obvious calcium carbonate dissolution circle around the colony, and pick the corresponding single colony to the MRS solid medium with the inoculation needle Three districts are cross-marked until the single bacterial colony that occurs is uniform in size, and Gram staining is selected as purple (positive), and the single bacterial colony with uniform shape and size of the stained bacterium is initially confirmed as lactic acid bac...

Embodiment 2

[0049] The broad-spectrum antibacterial activity of embodiment 2 Lactobacillus salivarius (L.salivarius) GZPH2

[0050] (1) Preparation of 39 strains of indicator bacteria

[0051] Inoculate a single colony of 39 strains of indicator bacteria (Table 1) into the nutrient broth liquid medium, culture at 37°C, 150r / min for 8h, and adjust its concentration to 1×10 6 CFU / mL, stored at 4°C, for later use;

[0052] Table 1 39 different experimental indicator strains

[0053]

[0054]

[0055] (2) Preparation of lactic acid bacteria

[0056] Inoculate the single colony of Lactobacillus salivarius (L.salivarius) GZPH2 identified in Example 1 in 50mL MRS liquid medium, 37°C, 150r / min, cultivate for 12h, and cultivate its concentration to 1×10 8 CFU / mL; 10000RCF, 4°C, centrifuge the lactic acid bacteria culture solution for 10 minutes, take the supernatant for the antibacterial test, store at 4°C, and set aside;

[0057] (3) (Supernatant) antibacterial activity screening

[00...

Embodiment 3

[0067] Other characteristic analysis of embodiment 3 Lactobacillus salivarius (L.salivarius) GZPH2

[0068] (1) Hemolysis analysis

[0069] Get 5 μ L seed liquid (final concentration is 8.0 * 10 8 CFU / mL) was spotted on a blood plate (sheep blood, Guangdong Huankai Biological Co., Ltd.), cultured at 37°C for 24 hours, and observed whether there was hemolysis around the colony. Select No. 8 vibrio parahaemolyticus (Vibrio parahaemolyticus) as positive control;

[0070] (2) Histamine detection

[0071] Prepare nutrient broth liquid medium, containing mixed amino acids (histidine 10.0g / L), adjust the pH of the medium to 5.5, add bromocresol purple (0.06g / L), agar powder (20g / L, gel strength >1300g / cm 2 ), sterilized at 120°C for 15min and poured onto the plate; activate the identified Lactobacillus salivarius (L.salivarius) GZPH2 bacterial strain in Example 1, after appropriate dilution, draw 100 μ L to coat the plate, cultivate it at 37°C for 48h, and make a positive control...

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Abstract

The invention belongs to the field of microbes, and concretely relates to a lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof. The lactic acid bacterium with wide-spectrum bacteriostatic activity is named lactobacillus salivarius GZPH2, is preserved at China Center for Type Culture Collection (CCTCC) since 27th, November, 2014, and has the preservation number of CCTCC M 2014598. The provided lactic acid bacterium lactobacillus salivarius GZPH2 has good bacteriostatic effect on multiple pathogenic bacteria of human and aquaculture bionts, and possesses the characteristics of not generating histamine, no hemolysis, acid resistance, cholate resistance and the like.

Description

technical field [0001] The invention belongs to the field of microorganisms, and in particular relates to a lactic acid bacterium with broad-spectrum antibacterial activity and application thereof. Background technique [0002] Vibrio (Vibrio) is one of the most common bacterial groups in the marine environment, widely present in seawater, seabed sediments, and seafood such as fish, shrimp, shellfish, crabs, sea cucumbers, and kelp. The 8th edition of "Bergey's Handbook of Bacteriology" recorded 35 species of Vibrio, a considerable part of which are important pathogenic bacteria of aquaculture organisms. In the aquaculture industry, the diseases of cultured organisms caused by Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio anguillarum, etc. are collectively called vibriosis, which causes huge economic losses to the aquaculture industry every year. At the same time, some Vibrios are also pathogenic bacteria of humans. They can not only cause gastroenteritis in humans...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K35/744A61P31/04C12R1/225
CPCA61K35/744C12N1/205C12R2001/225
Inventor 蔡俊鹏郭衍彪扶雄
Owner SOUTH CHINA UNIV OF TECH
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