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A kind of microbacterium in intertidal zone and its application

A microbacillus and intertidal zone technology, applied in the field of microorganisms, to achieve the effect of ensuring food safety

Active Publication Date: 2018-09-07
GUANGDONG OCEAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the use of microorganisms in shrimp paste to degrade T-2 toxin

Method used

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  • A kind of microbacterium in intertidal zone and its application
  • A kind of microbacterium in intertidal zone and its application
  • A kind of microbacterium in intertidal zone and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1 Separation and purification of T-2 toxin-degrading microorganisms

[0019] The preparation of traditional shrimp paste: the commercially available fresh shrimps are selected, washed and drained, after beating, add 30% of the shrimp weight in salt, stir well and divide into two parts, one part is used as a sample of fresh shrimp paste, and the other part is submerged in a fermenter for fermentation Naturally fermented, exposed to the sun and dew at night, stirred twice a day, 20 minutes each time, and fermented for 20 days as a sample of fermented shrimp paste.

[0020] Take 2.00 g of the fresh shrimp paste and fermented shrimp paste samples above, divide the two batches of samples into 4 groups, add 0, 10, 20, and 30 ng of T-2 toxin, and carry out micro-fermentation under the same conditions. After 1d, 5d, 10d, 15d, and 20d of fermentation, LC-MS / MS was used to detect the residual amount of T-2 toxin in the fermentation broth.

[0021] Degradation rate of T-2...

Embodiment 2

[0023] Identification of the isolated bacterial strain of embodiment 2

[0024] The bacterial strain CAMT25481 isolated in Example 1 was analyzed for biochemical characteristics using the VITEK 2 Gram-negative Bacteria Identification Card, and referred to the "Bergery Bacteria Identification Manual". Phylogenetic analysis and strain identification were carried out with 16S rRNA analysis method.

[0025] The culture characteristics of strain CAMT25481 are: young cultures are bacillus, old cultures are spherical, 1.1~1.2μm×1.4~3.2μm, Gram positive; move with perinatal flagella, facultative anaerobic; on nutrient agar The colony is flat, light orange, and the pigment does not spread; it is not sporulating, not acid-resistant, and salt-tolerant; alkaliphilic, can grow at pH6.5-11.5; chemoheterotrophic bacteria, fermentative metabolism.

[0026] Biochemical properties of strain CAMT25481: producing β-glucuronidase, α-glucosidase, pyrrolidinyl arylase, tyrosine arylase, β-xylosidas...

Embodiment 3

[0033] Example 3 Test of Degradation Ability of Microbacterium CAMT25481 in the Intertidal Zone to T-2 Toxin

[0034] The pure culture of the above strain CAMT25481 was inoculated into nutrient broth NB containing 5, 10, 15, 20, 25 ng / mL T-2 toxin, wherein the concentration of the strain CAMT25481 was 10 5 -107 CFU / mL; vortex for 60s, culture in a full-temperature shaker cabinet at 30±1°C, 180r / min, after 72 hours, draw 2mL of NB enrichment solution into a centrifuge tube, add 2mL of ethyl acetate, vortex for 60s, and sonicate for 10min , take the supernatant in another centrifuge tube (if the stratification is not obvious, centrifuge at 4000r / min for 5min), repeat the extraction 3 times, and combine the extracts. The extract was blown dry at 60°C under a nitrogen blower. Aspirate 2mL methanol aqueous solution (V / V:3 / 7) to dissolve, vortex for 30s, and sonicate for 5min. Pass through a 0.22 μm filter membrane, detect the T-2 toxin content by LC-MS / MS, and record the experime...

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Abstract

The invention belongs to the technical field of microorganisms, and particularly discloses intertidal Exiguobacterium aestuarii and application thereof. The strain is the intertidal Exiguobacterium aestuarii CAMT25481, the strain is preserved in China General Microbiological Culture Collection Center (CGMCC) on Oct.19th, 2015, and the preservation number of the strain is CGMCC NO. 11512. The degradation rate of the strain CAMT25481 to T-2 toxin can reach 73.80 percent to the maximum extent, the T-2 toxin can be effectively degraded, the significant problems that feed and raw material shrimps are polluted by the T-2 toxin and cannot be utilized, and environmental pollution is severe are solved, and a foundation is laid for ensuring the food safety of highly processed products of the raw material shrimps.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a microbacterium in the intertidal zone and its application. Background technique [0002] T-2 toxin is a sesquiterpene mycotoxin produced by a variety of Fusarium fungi. It is the most toxic toxin in the trichothecene family of toxins. 9 -C 10 Inter-double bond, hydroxyl, and acetoxy, the toxicity is mainly manifested as cytotoxicity and immune system toxicity, and has the "three-cause" effects of teratogenic, carcinogenic, and mutagenic, and has a potential carcinogenic threat to human and animal health. T-2 toxin is ubiquitous in aquatic feed, and prawns have strong tolerance to mycotoxins, so T-2 toxin can easily accumulate in prawns. In addition, the pure product of T-2 toxin is a white needle-like crystal, insoluble in water, easily soluble in polar organic solvents such as methanol, absolute ethanol, ethyl acetate, acetone, etc., stable in nature, strong in heat r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L5/20C12R1/01
Inventor 王雅玲孙力军谢主兰刘颖黄正花
Owner GUANGDONG OCEAN UNIVERSITY
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