Bifidobacterium adolescentis bacteriocin as well as production method and special production strain of bifidobacterium adolescentis bacteriocin

A technology of bifidobacteria and bacteriocin, applied in the field of food biology, to achieve good food processing characteristics and safety, and the effect of a wide antibacterial spectrum

Active Publication Date: 2014-09-17
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no reports of bacteriocin-producing Bifidobacterium adolescentis at home and abroad.

Method used

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  • Bifidobacterium adolescentis bacteriocin as well as production method and special production strain of bifidobacterium adolescentis bacteriocin
  • Bifidobacterium adolescentis bacteriocin as well as production method and special production strain of bifidobacterium adolescentis bacteriocin
  • Bifidobacterium adolescentis bacteriocin as well as production method and special production strain of bifidobacterium adolescentis bacteriocin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Isolation and Identification of Bifidobacterium adolescentis BL-8CGMCC No. 7791

[0024] 1. Screening of bacterial strains

[0025] Aseptically weigh 25g of the feces of the long-lived elderly in Turpan, Xinjiang, put it into 225mL of buffered peptone water quickly, and then dilute it 10 times to the number of viable bacteria 10 -6 -10 -10 cfu / mL, and then draw 0.2mL for Hengate anaerobic roller tube, and put it in a 37°C incubator for 48-72h. Select small and medium-sized colonies characterized by milky white or slightly yellowish, neat edges, convex round, smooth, and soft texture, and inoculate them in the modified MRS liquid medium, culture anaerobically for 24-48 hours, and perform Gram staining. Select colonies with morphological characteristics of bifidobacteria observed by Gram staining, white or yellowish, smooth and neat edges, and transparent circles around them, and inoculate them in the modified MRS medium, and culture them in aerobic and anaer...

Embodiment 2

[0032] Embodiment 2, Utilize Bifidobacterium adolescentis (Bifidobacterium adolescentis) BL-8CGMCC No. 7791 to produce bacteriocin

[0033] 1. Detection method of bacteriocin activity

[0034] (1) select standard curve range: 500mg nisin (nisin) standard substance (10 6 AU / g) was dissolved in 50mL sterile 0.02mol / L dilute hydrochloric acid solution to make 10 4 AU / mL standard solution for use. With 0.02mol / L HCl successively make 10 4 The AU / mL nisin standard solution was diluted into 5000, 2500, 1000, 750, 500, 250, 100, 75, 50, 25, 10 AU / mL standard solutions. Take 100 μL respectively and add them to the Oxford cups that have prepared the plates in turn, repeat 3 plates for each concentration, and do the antibacterial test, take the correction value of the diameter of the inhibition zone as the abscissa, and the logarithm of the titer of nisin as the ordinate, Draw a standard curve. Determine the nisin potency range when the diameter of the inhibition zone has a good li...

Embodiment 3

[0058] Example 3, Extraction and purification of bacteriocin produced by Bifidobacterium adolescentis BL-8CGMCC No. 7791

[0059] 1. Extraction by ammonium sulfate salting out precipitation method

[0060]Determination of optimal ammonium sulfate saturation: with the optimal medium composition and culture conditions obtained in Case 2, culture Bifidobacterium adolescentis BL-8CGMCC №.7791, and obtain the fermented liquid at 4°C, 8000rpm Centrifuge for 10 min, retain the supernatant, add 20-80% ammonium sulfate, stir for 2 h, and place overnight at 4°C. Take out and centrifuge at 12000rpm at 4°C for 10min, redissolve the precipitate in 1 / 5 of the original volume of the fermentation broth in 25mM phosphate buffered saline PBS (pH6.5) to obtain the crude extract of bacteriocin, and use PBS with the same saturation of ammonium sulfate as the control . Finally, the obtained bacteriocin crude extract was dialyzed with a 1000Da dialysis bag to desalinate, then concentrated to the o...

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Abstract

The invention discloses bifidobacterium adolescentis bacteriocin as well as a production method and a special production strain of the bifidobacterium adolescentis bacteriocin. The bifidobacterium adolescentis bacteriocin provided by the invention is obtained by fermenting bifidobacterium adolescentis BL-8 CGMCC No.7791. The invention provides an optimal culture medium component and a culture condition of bacteriocin generated by metabolism of bifidobacterium adolescentis and also provides an extraction and purification method of the bifidobacterium adolescentis bacteriocin. The extraction and purification method comprises the steps of: crude extraction through ammonium sulfate salting and precipitating; gel filtration chromatography; cation exchange resin chromatographic purification to obtain pure bacteriocin. The bifidobacterium adolescentis bacteriocin provided by the invention has the molecular weight of 829.23Da, is novel bifidobacterium adolescentis bacteriocin, is very good in food processing property, wide in antibacterial spectrum and high in safety and can be used for developing natural biological preservatives for foods.

Description

technical field [0001] The invention relates to a bifidobacterium adolescentis bacteriocin, a production method thereof and a special production strain, and belongs to the field of food biotechnology. Background technique [0002] In the process of processing, transportation and storage, food is extremely susceptible to microbial contamination, corruption and deterioration, which can lead to various food safety incidents. The development of new green food preservatives is one of the important problems that the food industry urgently needs to solve. In recent years, it is non-toxic, has no side effects, The emergence of natural microbial preservatives without drug resistance and degradable in the human body provides a new idea to solve this problem. [0003] Lactic acid bacteria bacteriocin is a polypeptide or protein substance with antibacterial activity produced by lactic acid bacteria through the ribosome synthesis mechanism during the metabolic process, which can inhibit ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/00C07K1/36C07K1/30C07K1/18C07K1/16C12R1/01
Inventor 刘国荣任丽王成涛宋振芹苏航
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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