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Strain of bacillus amyloliquefaciens, and antifungal protein and prawn fresh-keeping application thereof

A technology of amylolytic spores and antibacterial proteins, which is applied in the direction of bacteria, chemical instruments and methods, and methods based on microorganisms, and can solve problems that affect the development of the seafood industry, spoilage of shrimp bodies, and high shrimp spoilage rates.

Active Publication Date: 2017-03-29
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high protein and high water content of the prawn, it is easy to spoil under the action of microorganisms and enzymes after harvesting. In addition to the limitations of the logistics industry, the spoilage rate of the prawn is very high, which affects the development of the seafood industry.
[0012] The muscles, body fluids and viscera of fresh and alive prawns are all sterile, and the bacteria in the intestinal tract are also in a state of mutual restraint and balance. However, the part of the body surface that is in direct contact with the water body is attached to a variety of microorganisms, and they gradually move inward after the shrimp dies. Invasion, decomposition of nutrients, and eventually lead to spoilage of shrimp body

Method used

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  • Strain of bacillus amyloliquefaciens, and antifungal protein and prawn fresh-keeping application thereof
  • Strain of bacillus amyloliquefaciens, and antifungal protein and prawn fresh-keeping application thereof
  • Strain of bacillus amyloliquefaciens, and antifungal protein and prawn fresh-keeping application thereof

Examples

Experimental program
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Embodiment 1

[0046] The screening and identification of Bacillus amyloliquefaciens PJ comprises the following steps:

[0047] 1. Sudden death of live Penaeus vannamei shrimp, rinse with running water, and take the shrimp intestine; weigh 1g of shrimp intestine, cut it into pieces, soak it in 9ml of normal saline, and make the stock solution of the bacterial suspension;

[0048] 2. Gradiently dilute the stock solution of the bacterial liquid suspension, select the concentration of the bacterial liquid with a McFarland turbidity gradient of 0.5 and mix it evenly with the NA medium, and after cooling, cultivate it in a 30°C incubator for 24 hours; the formula of the NA medium is: beef Cream 3g, peptone 10g, NaCl 5g, agar 20g, water 1000ml, pH value 7.0-7.2;

[0049] 3. Pick the strains with rough surface and wrinkles, white opaque, round or irregular shape, etc. that meet the characteristics of the Bacillus amyloliquefaciens colony, carry out Gram staining, and observe through the microscope ...

Embodiment 2

[0070] Isolate and prepare antibacterial protein from the fermentation broth of Bacillus amyloliquefaciens PJ, comprising the following steps:

[0071] After activation of Bacillus amyloliquefaciens PJ, pick one ring and inoculate it in 100mL NA liquid medium, and culture it under shaking at 25-37°C and 150-200r / min for 8 hours to prepare a seed fermentation liquid; take the seed liquid and inoculate it at a pH value of In the basic fermentation medium of 6.5-7.2, the inoculum size is 3-6% (V / V), and the bacteria are shaken at 28-37°C and 150-200r / min for 12-18h. After the fermentation is completed, the fermentation broth is centrifuged at 10000r / min for 20min to obtain a supernatant, and the supernatant (that is, containing the antibacterial protein) is concentrated to 10ml to be the supernatant concentrate. Add ammonium sulfate to 20ml of the supernatant to 40-100% saturation, let it stand overnight, then centrifuge to collect the precipitate, and vacuum freeze-dry the preci...

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Abstract

The invention discloses a strain of bacillus amyloliquefaciens, and an antifungal protein and a prawn fresh-keeping application thereof. The bacillus amyloliquefaciens PJ has been preserved in Guangdong Culture Collection Center (GDMCC) on September 12nd, 2016. The address of GDMCC is fifth floor, No.59 building, courtyard, Xianlie middle road No.100, Guangzhou, Guangdong province. The preservation number is GDMCC No. 60076. The provided novel strain and antifungal protein thereof have a biological activity on preventing pseudomonas, micrococcus luteus, shewanella spp., and acinetobacter, can be applied to the fields such as health care product, food fresh-keeping, cosmetic preservative, and the like, and have a good market potential.

Description

technical field [0001] The invention belongs to the field of biological preservation, and in particular relates to a strain of bacillus amyloliquefaciens and its antibacterial protein and the application thereof in preservation of prawns. Background technique [0002] Food preservation refers to the method of using physical, chemical and biological means to process food, so as to slow down its spoilage speed as much as possible and maintain its original freshness. [0003] Due to the high protein and high moisture characteristics of seafood, it is easy to spoil during storage and transportation. Therefore, it is urgent to update the preservation technology of seafood. Common fresh-keeping methods for seafood include low-temperature preservation, modified atmosphere preservation, irradiation preservation, chemical preservation and biological preservation. [0004] Refrigeration and preservation are divided into four preservation methods: refrigeration, ice temperature, sligh...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P21/02C07K14/32A23B4/22C12R1/07
CPCA23B4/22C07K14/32C12N1/205C12R2001/07
Inventor 黄儒强吴佩君朱宝君汪光华杨喆王静辉马广智陈祥茂陈珂王倩高林林
Owner SOUTH CHINA NORMAL UNIVERSITY
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