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VBNC state Lactobacillus brevis CSHRR5-3 strain and application thereof

A kind of Lactobacillus brevis, strain technology, applied in the field of microorganisms, can solve problems such as lack of enzyme production system

Active Publication Date: 2021-08-06
ZHEJIANG NORMAL UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Studies have shown that insects lack a complete enzyme production system and need to rely on intestinal bacteria to provide a variety of enzymes to complete food digestion, nutrient absorption, material metabolism and body immune function

Method used

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  • VBNC state Lactobacillus brevis CSHRR5-3 strain and application thereof
  • VBNC state Lactobacillus brevis CSHRR5-3 strain and application thereof
  • VBNC state Lactobacillus brevis CSHRR5-3 strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, bacterial strain CSHRR5-3 screening

[0042] (1) The samples were collected from the fresh silkworm feces of the 5th instar of Guican No. 2 in Wuping Township (23°14'N, 106°22'E) in Jingxi City, Guangxi in August 2018. The isolation of VBNC lactic acid bacteria was based on recovery promotion Factor (Rpf) is obtained by MPN method plus dilution coating plate separation, and the specific steps are as follows:

[0043] 1) Preparation of culture supernatant containing Rpf active protein, with reference to CN102618464B "A kind of resuscitation medium that can promote the growth of VBNC bacteria and improve the separation abundance and its preparation method and application", Micrococcus luteus (Micrococcusluteus) IAM14879 T Cultivate in liquid medium to the late logarithmic phase, filter the supernatant, filter it with a 0.22 μm filter membrane, and use it as an active Rpf protein solution; the active Rpf protein solution is sterilized at 121°C for 20 minutes ...

Embodiment 2

[0063] Embodiment 2, identification of bacterial strain CSHRR5-3

[0064] Morphological identification: After being cultured on the MRS medium plate for 24 hours, the single colony formed is round, raised, with smooth edges, milky white, moist, opaque, and 1-3mm in diameter ( figure 1 ), the bacteria are rod-shaped, Gram-positive, capsulated, without flagella, and without spores ( figure 2 ).

[0065] Physiological and biochemical characteristics test: The physiological and biochemical characteristics of the strain CSHRR5-3 were determined by referring to the culture medium recommended in the "Common Bacteria Identification Manual", and it was found that the strain can use glucose, maltose and lactose to grow; produce acid and gas on the glucose fermentation medium The methyl red test is positive; the nitrate is not reduced, the gelatin is not liquefied, the hydrogen sulfide test, urea test, V-P test, catalase and oxidase are all negative, Simon's citrate cannot be used, and...

Embodiment 3

[0070] Embodiment 3, the measurement of bacterial strain CSHRR5-3 growth curve and acid production ability

[0071] Take 1 ring of bacterial lawn of strain CSHRR5-3, inoculate it into a 250mL Erlenmeyer flask filled with 50mL of MRS liquid medium, and cultivate it in a shaker at 37°C and 140r / min for 18-22h as a seed solution. With 1% (volume ratio) inoculum amount, inoculate the seed liquid into the Erlenmeyer flask containing 50mL MRS liquid medium, shake and cultivate in a shaker at 37°C and 140r / min for 48h, and take a small amount of bacterial suspension every 2h Determination of OD value and pH value, the growth curve and pH change curve are shown in Figure 4. It can be seen from the growth curve that the strain enters the logarithmic phase of growth at 8 hours, and enters the stable growth phase after 30 hours. The pH value dropped to 3.14 at 40h, and stabilized at around 3.14 at 40-48h, indicating that the strain CSHRR5-3 had a strong acid-producing ability.

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Abstract

The invention discloses Lactobacillus brevis CSHRR5-3, and the preservation number is CGMCC No. 18475. The Lactobacillus brevis CSHRR5-3 has the following properties: the strain has strong acid production performance; a strain fermentation broth has antibacterial properties, and the strain fermentation broth has thermal stability, ultraviolet stability, acid-base stability and protease stability; the strain has gastric acid resistance and cholate resistance; and the strain can produce exopolysaccharides, and the exopolysaccharides have antioxidant activity. The invention also provides application of the Lactobacillus brevis CSHRR5-3 as a bacteriostatic agent and an antioxidant.

Description

technical field [0001] The invention belongs to the technical field of microbes, and in particular relates to a resuscitated cultureable Lactobacillus brevis (Lactobacillus brevis) CSHRR5-3 in VBNC state with good antibacterial effect and strong acid production performance and its application. Background technique [0002] Lactic acid bacteria is a general term for a class of non-spore-forming, Gram-positive bacteria that can ferment sugars to produce lactic acid [1] , is a kind of widely used probiotics, often used in fermentation industry and probiotics. The culturable microorganisms that can be isolated from most natural environment samples are only about 0.01% to 10%, and the vast majority of microorganisms have not been effectively identified because they are in a viable but non-culturable state (viable but non-culturable, VBNC). Bacteria is an effective way to discover new strain resources and develop environmental functional bacteria [2] . Studies have shown that t...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/747A61P31/04A61P39/06C12R1/24
CPCA61K35/747A61P31/04A61P39/06Y02A50/30
Inventor 张萍华苏盼许梦洁王星晨张艳军于红丁林贤谢乃钧
Owner ZHEJIANG NORMAL UNIVERSITY
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