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Alcaligenes faecalis and application thereof in degrading ochratoxin A

A kind of technology of Alcaligenes faecalis and microbial cells, applied in the field of Alcaligenes faecalis

Active Publication Date: 2016-08-17
ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the degradation of ochratoxin A by Alcaligenes faecalis

Method used

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  • Alcaligenes faecalis and application thereof in degrading ochratoxin A
  • Alcaligenes faecalis and application thereof in degrading ochratoxin A
  • Alcaligenes faecalis and application thereof in degrading ochratoxin A

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation, purification and identification of Alcaligenes faecalis ASAGF-OD1

[0027] Soil samples were collected from the pilot test base of the Beiqijia National Grain Bureau Science Research Institute in Changping District, Beijing. The 96-well deep-well plate was used as the culture carrier, and the micro-enrichment culture method was adopted. The samples were first made into bacterial suspensions in sterile water. solution, inoculated in OTA-LB liquid medium with 10% inoculation amount, the final concentration of OTA in the medium was 1 μg / ml, and transplanted with 10% inoculum amount after culturing for 7 days, and the OTA concentration in the medium remained unchanged. After 5 consecutive transplants, the OTA content was detected, and the OTA-LB medium containing the same OTA concentration without inoculation was used as a negative control. Spread the bacterial suspension with degraded OTA on the LB agar plate at an appropriate dilution according to the...

Embodiment 2

[0032] The 0D-1 strain of the present invention has been preserved on January 25, 2016 in the General Microbiology Center (CGMCC) of China Microbiological Culture Collection Management Committee (CGMCC) (address: No. 3, No. 1, Beichen West Road, Chaoyang District, Beijing, Microbiological Research, Chinese Academy of Sciences institute, zip code 100101), and its deposit number is CGMCC No.12100. Example 2 Detection method of ochratoxin A and ochratoxin alpha

[0033] HPLC-FLD detection method:

[0034] The detection method refers to Bragulat et al. (Bragulat, M., Abarca, M., & F. (2001). An easy screening method for fungi producing ochratoxin A in pureculture. Int J Food Microbiol, 71 (2), 139-144.) method and improvement. The details are as follows: draw 600 μL of sample, centrifuge at 14000 r / min for 10 min, take the supernatant and filter it with a 0.22 μm filter membrane, analyze it with HPLC-FLD or store the sample at 4°C.

[0035] Detection conditions: Agilent 5TC-C1...

Embodiment 3

[0039] Example 3 Alcaligenes faecalis ASAGF-OD1 is used to degrade ochratoxin A

[0040] Alcaligenes faecalis ASAGF-0D1 culture: Pick a single colony of Alcaligenes faecalis ASAGF-0D1 (preservation number: CGMCC No.12100), inoculate it in 3mL LB liquid medium, culture at 30°C, rotate at 220r / min, and cultivate Time 30h. After the fermentation, the fermentation broth was stored at 4°C for later use.

[0041] Take 50 μL of the prepared Alcaligenes faecalis ASAGF-OD1 fermentation broth and inoculate it in 50 mL of OTA-LB medium. The culture temperature is 30 and 37 °C. Samples are taken every 12 hours to measure the OTA content and calculate the detoxification rate. The experiment was repeated at least 3 times.

[0042] The formula of LB medium: tryptone 10g, yeast extract powder 5g, sodium chloride 10g, agar 1.5%, add distilled water to 1L, pH7.0-7.2, sterilize at 121°C for 20min.

[0043] Among them, the preparation method of OTA-LB medium: transfer a certain volume of ochra...

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Abstract

The invention discloses alcaligenes faecalis and application thereof in degrading ochratoxin A. The alcaligenes faecalis ASAGF-0D1 has the preservation number of CGMCC No. 12100. The invention also provides a biological detoxication agent containing a preserved strain and a preparation method of the biological detoxication agent. In addition, the invention also provides the application of the strain or the biological detoxication agent in degrading the ochratoxin A.

Description

technical field [0001] The invention relates to a strain of Alcaligenes faecalis, which can be used to degrade ochratoxin A in food or feed. Background technique [0002] Ochratoxin (Ochratoxin A, OTA) is a toxic secondary metabolite produced by fungi, widely present in food and feed, with nephrotoxicity, carcinogenicity, teratogenicity, neurotoxicity and immunotoxicity, etc., harmful to animals and There is great harm to human health. [0003] The main measures to control OTA pollution in feed and food are physical, chemical and biological detoxification techniques. Physical methods such as adsorption, heating, and radiation cannot effectively control the OTA content in food or feed. The chemical method can effectively degrade and transform OTA mainly through oxidation, acid and alkali, etc., but the nutritional content is greatly destroyed, and it causes uncertainty about the health hazards of chemical reagent residues. In addition, physical and chemical methods are als...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12P1/04A23L3/3571A23L5/20C12R1/05
CPCA23L3/3571C12P1/04C12N1/205C12R2001/05
Inventor 张晓琳张宏海汪洋赵晨
Owner ACAD OF NAT FOOD & STRATEGIC RESERVES ADMINISTRATION
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