Luteimonas sp for degrading alflatoxin B1 and ochratoxin A and application of luteimonas sp

A technology of Luteomonas luteus and aflatoxin, which is applied in the field of highly efficient degradation of aflatoxin B1 and ochratoxin A, and can solve the problems of no substantial detoxification significance and rare reports

Active Publication Date: 2016-01-20
江苏奥迈生物科技有限公司
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  • 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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  • Luteimonas sp for degrading alflatoxin B1 and ochratoxin A and application of luteimonas sp
  • Luteimonas sp for degrading alflatoxin B1 and ochratoxin A and application of luteimonas sp
  • Luteimonas sp for degrading alflatoxin B1 and ochratoxin A and application of luteimonas sp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Screening, identification and cultivation of aflatoxin B1 and ochratoxin A degrading bacteria

[0024] 1. Screening of bacterial strains

[0025] 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

[0035] Example 2 Degradation characteristics of bacterial strain CW574 detected by high performance liquid chromatography

[0036] Degradation dynamic detection of degrading strains: The degrading strain CW574 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.

[0037] The degraded supernatant was passed through the OTA immunoaffinity column and the AFB1 immunoaffinity...

Embodiment 3

[0042] Example 3 Application of bacterial strain CW574 in detoxification treatment of corn soybean meal type feed

[0043] 1. Experimental materials

[0044] 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.

[0045] Strain activation medium II: peptone 5.0g / L, beef extract 30.0g / L, NaCl 5.0g / L, pH7.0-7.2.

[0046] The above-mentioned medium was autoclaved at 120°C for 15 minutes, and the experimental feed was a corn-soybean meal-based diet.

[0047] 2. Experimental method

[0048] 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 luteimonas sp for degrading alflatoxin B1 and ochratoxin A and application of the luteimonas sp and particularly provides a strain of difunctional efficient degrading bacteria (Luteimonas sp.) CW574 and application thereof in degrading low-pollution-concentration alflatoxin B1 and ochratoxin A. Compared with existing aflatoxin degrading bacteria, the strain CW574 can achieve excellent degrading effects under the condition of low-concentration toxin pollution. Under the condition of liquid fermentation, in a fermentation culture solution containing alflatoxin B1 and ochratoxin A with the final concentration being 20 micrograms per liter, the degrading rate of the strain CW574 on ochratoxin A is 90.1% in 48 h; the degrading rate of the strain CW574 on alflatoxin B1 is 85.5% in 48 h. When the strain CW574 is used for treating fodder (the final concentration is 20 micrograms per kg) polluted by toxins, the degrading rate on OTA is 48.3% in 48 h, and the degrading rate on AFB1 is 52.1% in 48 h. The strain CW574 has substantive application value and significance on the application aspects of food and feed biological detoxification.

Description

technical field [0001] The invention relates to the field of microbiology and biodegradation, in particular to a bacterium Luteus luteus capable of efficiently degrading 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23L5/20A23K10/16C12R1/01
Inventor 周育王旭姜楠韦朝领邬艳博
Owner 江苏奥迈生物科技有限公司
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