Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of Lysobacter

A technology of aflatoxin and ochratoxin, applied in the field of microbiology and biodegradation, can solve the problems of rare reports and no substantial detoxification significance

Active Publication Date: 2016-01-27
ANHUI AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the virus-free strains reported so far have the following two common problems in practical application: First, the detoxification mechanism of some virus-free strains belongs to physical adsorption, not substantial biodegradation, and is adsorbed on the cell wall of the bacteria detoxification may occur under different physical and chemical environments in animals or humans, and has no substantial detoxification significance; second, the detoxification rate of the reported strains to toxins is mostly measured at a higher concentration of 100 μg / kg or more. There are few reports on the actual detoxification ability of low-concentration aspergillus toxoids (less than 20 μg / kg) in complex matrices such as food raw materials and feedstuffs

Method used

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  • Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of Lysobacter
  • Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of Lysobacter
  • Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of Lysobacter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Screening, identification and cultivation of aflatoxin B1 and ochratoxin A degrading bacteria

[0023] 1. Screening of bacterial strains

[0024] The samples were collected from soil heavily polluted by PAHs (near oil refineries, auto repair shops) and moldy food heavily polluted by mycotoxins. Take 0.5 g of the collected sample and add it to 150 mL of enrichment medium (the contents of OTA and AFB1 are about 20 μg / L each), and enrich and culture in a constant temperature incubator at 30°C for 7 days. After the first enrichment culture, inoculate 5 mL of the enrichment culture solution into 150 mL of fresh OTA and AFB1 enrichment medium, increase the concentration of OTA and AFB1 to 30 μg / L, and continue the enrichment culture under the same conditions for 7 sky. Complete the third enrichment culture experiment with the same enrichment method, increasing the concentration of OTA and AFB1 to 50μg / L. After the enrichment culture was completed, the enriched cu...

Embodiment 2

[0034] Example 2 Degradation characteristics of bacterial strain CW239 detected by high performance liquid chromatography

[0035] Degradation dynamic detection of degrading strains: The degrading strain CW239 was continuously activated for 2 generations on the most suitable solid medium, inoculated in 4mL liquid medium for overnight culture, and fresh bacterial liquid was obtained. Inoculate 50 μL of fresh bacteria liquid into 4mL LOTA test medium (containing 20.0 μg / LOTA) and AFB1 test medium (containing 20.0 μg / LAFB1) respectively, shake the inoculated test tube for 0h, 12h, 24h and 48h, and degrade each The experiment was set up with 3 repetitions. Escherichia coli K12 (E.coliK12) was used as a negative control strain. After the culture was completed, mix well, centrifuge at 8000r / min for 10min, and collect the supernatant and bacterial precipitate respectively.

[0036] The degraded supernatant was passed through the OTA immunoaffinity column and the AFB1 immunoaffinity...

Embodiment 3

[0041] Example 3 Application of bacterial strain CW239 in detoxification treatment of corn soybean meal type feed

[0042] 1. Experimental materials

[0043] Strain activation medium Ⅰ: tryptone 17.0g / L, soytone 3.0g / L, glucose 2.5g / L, NaCl 5.0g / L, K 2 HPO 4 2.5g / L, agar 20.0g / L.

[0044] Strain activation medium II: peptone 5.0g / L, beef extract 30.0g / L, NaCl 5.0g / L, pH7.0-7.2.

[0045] The above-mentioned medium was autoclaved at 120°C for 15 minutes, and the experimental feed was a corn-soybean meal-based diet.

[0046] 2. Experimental method

[0047] Add an appropriate amount of OTA and AFB1 standard stock solution to 50mL of phosphate buffer solution, mix it and immediately pour it into 100g of the crushed feed sample, stir evenly, so that the final concentration of OTA and AFB1 reaches 40.0μg / kg respectively, and feed in Dry in a cool and ventilated place for later use. Continuously activate the strains to be tested for 2 generations on the solid activation medium I...

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Abstract

The invention provides Lysobacter capable of efficiently degrading aflatoxin B1 and ochratoxin A and application of the Lysobacter and particularly provides double-function Lysobacter sp. CW239 and application thereof to the degradation of low-pollution concentration aflatoxin B1 and ochratoxin A. Compared with existing aflatoxin degrading bacteria, the Lysobacter sp. CW239 has the advantages that the Lysobacter sp. CW239 can achieve excellent degrading effect under a low-concentration toxin pollution condition; under a liquid fermentation condition, in fermentation broth, which contains the aflatoxin B1 and ochratoxin A, with the final concentration of 20 microgram/L, the 12-hour ochratoxin A degradation rate of the Lysobacter sp. CW239 is 53.1%, and the 48-hour degradation rate reaches 99.8%; the 12-hour aflatoxin B1 degradation rate of the Lysobacter sp. CW239 is 42.5%, and the 48-hour degradation rate reaches 83.4%; when the Lysobacter sp. CW239 is used for processing feed (with the final concentration of 20 microgram/kg) polluted by toxins, the 48-hour ochratoxin A degradation rate is 68.7%, and the 48-hour aflatoxin B1 degradation rate is 52.1%; the Lysobacter sp. CW239 has substantial application value and significance when being applied to food and feed bio-detoxification.

Description

technical field [0001] The invention relates to the fields of microbiology and biodegradation, in particular to a lysobacterium that efficiently degrades aflatoxin B1 and ochratoxin A and an application thereof. Background technique [0002] Aspergillus mycotoxins are mainly secondary metabolites produced by Aspergillus flavus, A. (OTA) is the most toxic and most widely polluted in agricultural production and food industry. In 1993, aflatoxin was classified as a Class I carcinogen by the World Health Organization (WHO) Cancer Research Institute. Because of its strong carcinogenic, teratogenic and mutagenic properties, aflatoxin has gradually become the focus of public health and scientific research. Contamination of grain and oil crops with aflatoxins and ochratoxins has become a global problem. In animal husbandry, the two types of aspergillus can endanger animal health by contaminating feed, leading to low productivity in animal husbandry, causing serious economic losses,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20A23K10/00C12R1/01
Inventor 王旭周育姜楠张正竹
Owner ANHUI AGRICULTURAL UNIVERSITY
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