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Mushroom rot-causing fungus bacteriophage and application thereof

A bacteriophage and bacterial liquid technology, applied in the field of microorganisms, can solve the problems of chemical reagents affecting the health of consumers, human deformities or cancer, etc., and achieve the effects of non-toxic side effects, inhibiting metabolism and reproduction, and reducing pollution.

Active Publication Date: 2021-07-30
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]Although the above preservation methods can effectively inhibit the microorganisms in edible fungi and prolong their shelf life, the residues of chemical reagents will also affect the health of eaters, and eating too much chemical Preservatives may cause deformities or cancer in humans

Method used

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  • Mushroom rot-causing fungus bacteriophage and application thereof
  • Mushroom rot-causing fungus bacteriophage and application thereof
  • Mushroom rot-causing fungus bacteriophage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Obtaining of bacteriophage BTXBG52

[0044] 1. Isolation and identification of spoilage bacteria XBG5 from edible fungi

[0045] (1) The edible fungi used in this stage are commercially available edible fungi, which are sealed and stored in sterile homogeneous bags, and placed in a constant temperature incubator at 37°C to accelerate corruption.

[0046] (2) Weigh 5 g of the above spoilage edible fungus samples and put them in a sterile homogeneous bag filled with 45 mL of sterile saline, and homogenize for 1-2 min with a homogenizer, which is 10 -1 The homogeneous sample was serially diluted with sterile saline. Use a pipette to take 1mL of each gradient dilution solution into a 90mm sterile plate, pour about 15-20mL of nutrient agar medium that has been melted and cooled to about 46°C, and immediately rotate the plate to fully mix the bacterial solution with the medium. After the culture medium was completely solidified, the plate was turned upside down a...

Embodiment 2

[0093] Embodiment 2: Drawing of one-step growth curve of phage BTXBG52

[0094] Take a ring of host bacteria cultured on a slant and inoculate it in a sterile LB liquid medium for 12-16 hours, then transfer it to a 100mL sterile LB liquid medium at a 3% inoculum size, at 37°C, 220r / h Min shaker culture to logarithmic growth phase (OD 600 ≈0.6), centrifuged at 4000r / min for 8min to collect the bacteria. Resuspend the bacteria in 500 μL of sterile liquid LB, add 500 μL of diluted phage particles to it to make MOI=10 -4 (MOI = phage titer / host bacterial concentration), after static adsorption for 1min, immediately centrifuge at 10000r / min for 1min and discard the supernatant to remove free phage particles. Immediately place it in a shaker at 37°C and 220r / min, take samples every 5 minutes, use the double-layer plate method to measure the titer of the phage in the process of 0-100 minutes, repeat three times at each time point to get the average titer, and draw a step growth cu...

Embodiment 3

[0097] Embodiment 3: Determination of bacteriophage BTXBG52 cracking ability

[0098] Pick a single colony of the host bacterium XBG5 on LB solid medium, inoculate it in 5mL sterile LB liquid medium, cultivate it on a shaker at 37°C and 220r / min for 12 hours, then serially dilute it 10 times, and measure it by plate coating method For the number of live bacteria, dilute the host bacteria liquid with sterile LB medium to make the bacterial concentration 10 7 cfu / mL. The purified phage lysate was amplified to a titer of 10 9 pfu / mL, the host bacterial liquid and phage lysate were mixed according to different MOI (MOI=phage titer / host bacterial colony number), and distributed in 96-well plates, and each MOI was set to 5 parallels. Place the 96-well plate at 37°C, 160r / min, shake and culture for 10h, and measure the OD of the culture solution with a microplate reader every 30min 600 , to OD 600 The average value is the ordinate, the time is the abscissa, and the antibacterial ...

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Abstract

The invention belongs to the technical field of microorganisms, and relates to a bacteriophage of enterobacter separated from rotten mushrooms and application of the bacteriophage as a biological additive component or a biological preservative in preservation of edible mushrooms. The mushroom rot-causing fungus acteriophage is specifically bacteriophage BTXBG52, and the preservation number is CGMCC NO.20708. The bacteriophage can specifically split enterobacter bacteria separated from rotten edible mushrooms, and has the advantages of strong thermal stability, high chloroform tolerance, large release amount and good preservative and fresh-keeping effects on edible mushrooms. Whole genome sequencing results show that the bacteriophage is free of virulence factors, and comparison results of BLAST homologous sequences in an NCBI database show that the homology of the bacteriophage genome and currently known bacteriophage strains in the database is only 9%. The bacteriophage is used for preparing a food bacteriostatic agent or a bacterial bacteriostatic agent, and the bacteriophage can be applied to preservation and fresh keeping of edible mushrooms, medical sterilization and factory production.

Description

Technical field: [0001] The invention belongs to the technical field of microbes, and relates to a bacteriophage isolated from enterobacteria in spoilage mushrooms and its application as a biological additive component or a biological fresh-keeping agent in the fresh-keeping of edible fungi. Background technique: [0002] The cells of edible fungi will die slowly after being picked, and the microorganisms attached to the fungus will multiply and grow rapidly. The mushroom body is rich in water and nutrients, which will cause the microorganisms to multiply in large numbers and quickly cause the fruiting body to rot and deteriorate. Santana et al. (Santana CC, VanettiMCD, Kaauya MCM. Microbial growth and color of minimally processed shitake mushroom stored at different temperatures. International Journal of Food Science and Technology, 2008, 43, 1281-1285.) studied how temperature changes affect the Microorganisms, the dominant spoilage bacteria during the storage process are ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A23B7/155A23L3/3571C12R1/92
CPCC12N7/00A23B7/155A23L3/3571A23V2002/00A23V2200/10
Inventor 贾士儒谭之磊杨洪江李欣颖李东航侯颖
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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