Heat-resisting type bifidobacterium longum, screening method thereof and preparation method for bacterial powder

A technology of Bifidobacterium longum and screening method, which is applied in the fields of heat-resistant Bifidobacterium longum and its screening method and bacterial powder preparation, and can solve the problems of inability to ensure product placement conditions, difficulty in maintaining high activity, and decline in storage stability, etc. problems, to achieve the effect of excellent stability of viable count, short screening cycle and slow downward trend

Inactive Publication Date: 2018-01-19
JIANGSU WECARE BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Probiotics generally have high requirements on the environment and poor tolerance to the outside world. It has been difficult to maintain high activity in long-term storage at room temperature, which limits product development and application.
Although the probiotics currently on the market are supplied in the form of freeze-dried bacteria powder, the storage stability of the probiotics generally shows a serious decline in the shelf life, especially for products stored at room temperature or at a high external temperature. Significant decline or sudden drop in the number of viable bacteria in a short period of time
[0004] At present, most domestic products of microecological preparations are stored at room temperature or at room temperature. In some areas or there are local periods of high temperature or long-term harsh weather, the storage conditions of the products cannot be guaranteed, or some products that require special processes need to go through high temperature for a long time. For example, in the manufacturing process of capsules, soft candies, chocolates and other products, the live bacteria in the products will be greatly lost, so a screening method that can improve the tolerance of the strain itself is needed to deal with the development and application of different products.

Method used

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  • Heat-resisting type bifidobacterium longum, screening method thereof and preparation method for bacterial powder
  • Heat-resisting type bifidobacterium longum, screening method thereof and preparation method for bacterial powder
  • Heat-resisting type bifidobacterium longum, screening method thereof and preparation method for bacterial powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1: Screening, isolation and cultivation of thermotolerant Bifidobacterium longum

[0046] Bifidobacterium longum was isolated from feces samples of adult healthy human beings, diluted aseptically, and the diluted solution was drawn and spread on the selective MRS medium, cultured under anaerobic conditions at 37°C for 48 hours, and the calcium-dissolving circle was selected to be larger. The colony was a milky white raised colony, and was examined under a microscope. After being purified and separated by multiple streaks, the growth temperature was initially screened at 25-45°C, and a total of 112 strains grew at a pH of 4.0-7.2, and were stored at 4°C.

[0047] The components of the diluent used are: 0.085% sodium chloride, 0.1% casein peptone, and the rest is water. The main components of the selective medium used are: casein peptone 10g, yeast extract powder 5g, dipotassium hydrogen phosphate 6g, ammonium citrate 2g, sodium acetate 25g, magnesium sulfate 0.57...

Embodiment 2

[0049] Example 2: Stress screening and strain identification by cyclic heat stimulation

[0050] The strains screened and stored at low temperature in Implementation 1 were expanded by MRS medium, the fermentation broth was centrifuged, the supernatant was poured, and 1-5mL of diluent was added, blown and mixed evenly, so that the bacteria sludge was fully dispersed in the diluent, and the After the bacteria-containing dilution is sealed, put it in a constant temperature water bath at 75°C for heat shock, and time the heat shock for 5-30 minutes. After the heat shock is completed, quickly use cooling water to cool it below 20°C, and spread the heat-shocked bacteria-containing dilution. Separation is done by streaking, and the well-growth and medium-sized colonies are selected for expansion in the MRS medium. The expanded fermentation broth is centrifuged, diluted, heat-shocked, coated, and screened for a series of cycles. The number of cycles is 50 times. Conditional screening...

Embodiment 3

[0058] Embodiment 3: Characteristic test of Bifidobacterium longum bacterial strain BL21

[0059] 1. Adhesion characteristic test

[0060] The adhesion of lactic acid bacteria to host intestinal epithelial cells is strain-specific rather than host-specific, so an in vitro model can be used to screen the desired lactic acid bacteria. The adhesion process of bifidobacteria is firstly through non-specific contact, and then binds to the receptor. In this experiment, according to the Bifidobacterium longum strain BL21 obtained in Example 2, its ability to adhere to the extracellular mechanism was tested, and it was found that it has High adhesion properties.

[0061] Centrifuge the bacterial liquid of Bifidobacterium longum BL21 at 8000r / min for 15 minutes, remove the supernatant, collect the precipitated bacterial cells, remix the bacterial cells and RPMI-1640 cell culture medium, and make the concentration reach 10 8 CFU / mL. Add the prepared bacterial suspension to a 24-well p...

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Abstract

The invention relates to the field of probiotics and especially relates to a heat-resisting type bifidobacterium longum, a screening method thereof and a preparation method for bacterial powder. The heat-resisting type bifidobacterium longum is preserved at China General Microbiological Culture Collection Center with the preservation number of CGMCC NO.10452. The screening method thereof comprisesthe steps of collecting of samples, conditional preliminary screening and secondarily screening for stress resistance. The preparation method thereof comprises the following steps: activating bacterial strain, expanding culture, anaerobically fermenting, collecting and emulsifying thalli, freeze-drying emulsion, crushing and then packaging. The heat-resisting type bifidobacterium longum providedby the invention has high activity, is capable of obviously promoting the stability within the shelf life under normal temperature or high temperature and has excellent acid resistance, cholate resistance and intestinal tract adhesion in human gastrointestinal tracts.

Description

technical field [0001] The invention relates to the field of probiotics, in particular to a heat-resistant bifidobacterium longum, a screening method thereof, and a preparation method of bacterial powder. Background technique [0002] Bifidobacterium longum belongs to probiotics that can be added to food, and mainly involves the application of food, health care products, daily chemical products, feed and other products. Probiotic powder, especially probiotic powder related to lactic acid bacteria, is common in most probiotic fermented foods, health product preparations, biological feed, and pharmaceutical preparations. [0003] Probiotics generally have high environmental requirements and poor tolerance to the outside world. It has been difficult to maintain high activity in long-term storage at room temperature, which limits product development and application. Although the probiotics currently on the market are supplied in the form of freeze-dried bacteria powder, the sto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12Q1/04C12R1/01
Inventor 方曙光陈珂可冯胜鹏姜甜朱建国
Owner JIANGSU WECARE BIOTECHNOLOGY CO LTD
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