Kluyveromyces marxianus C2 and preparation method and application thereof

A Kluyveromyces cerevisiae and seed technology, applied to Kluyveromyces marxianus C2, the preparation method and application field of the yeast, can solve the problems of non-listing and the like, achieve safe use, good environmental tolerance, strong inulin The effect of degrading enzyme activity

Active Publication Date: 2014-07-30
WUHAN KEQIAN BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The patent application number is 201310083469.1, and the name is "a preparation method and application of a zearalenone biodegradation agent", which provides a brewer's yeast extract that can be used for the biodegradation of zearalenone toxin; the above patent, at the same time De Vossia, which degrades a variety of mycotoxins, belongs to the genus De

Method used

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  • Kluyveromyces marxianus C2 and preparation method and application thereof
  • Kluyveromyces marxianus C2 and preparation method and application thereof
  • Kluyveromyces marxianus C2 and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A kind of acquisition of Kluyveromyces marxe C2, its steps are:

[0034] 1. Collect fresh feces from healthy pigs and soil samples from vegetable fields and ditches around the pig farms from Huangpi natural breeding soil pig farm in Hubei. Shake the samples with sterile water to evenly form a suspension. After gradient dilution of 1000, take 0.2ml and add 100ug of aflatoxin B1 was spread on the Martin Bengal red agar plate (purchased from Qingdao Haibo Biology), and a single colony was cultivated at 30°C. Pick different single colonies and streak on the Martin Bengal red agar plate for several times to obtain purified strains, verify the degradation effect on aflatoxin B1, and finally obtain a strain capable of degrading aflatoxin B1, add glycerol, and store in a -80°C refrigerator save. For this strain, it is defined as C2 strain.

[0035] 2. Through physiological, biochemical and 26s identification, it was finally determined that C2 was a strain of Kluyveromyces marxi...

Embodiment 2

[0038] A preparation method of Kluyveromyces marxe C2, the steps are:

[0039] A. Use an inoculation loop to pick the slanted seeds of the C2 strain, inoculate the YDP shake flask seed medium, and cultivate at 35°C for 18-20 hours as the primary seed solution.

[0040] The formula of YDP medium is (1L): glucose 20g, yeast powder 10g, peptone 20g; pH value 7.2, sterilized at 121°C for 20min.

[0041] B, the seed liquid of cultivating is inoculated in 50L fermenter by the inoculum amount of 1% (V / V) and enlarges culture, and fermentation medium filling liquid is 65%, supplements defoamer on-line in real time in the cultivation process, controls dissolved oxygen concentration Above 30%, temperature controlled at 32°C, cultivated for 24 hours, and the OD600 value reached about 10, used as the secondary seed solution.

[0042] C, by 3% (V / V) inoculum amount, the secondary seed liquid is inoculated into the fermentation medium of 10t fermenter tank, the amount of fermentation liqui...

Embodiment 3

[0046] C2 bacteria degradation aflatoxin B1 solution test:

[0047] Prepare aflatoxin sterile solution of 6000ng / ml, and add it to the following three test solutions according to the ratio of 1:9 (v / v), A 1 press 10 6 cfu / ml YPD fresh culture medium after inoculating Kluyveromyces marxe C2, B 1 press 10 6 cfu / ml inoculated with Kluyveromyces marxe C2 and cultured for 30 hours in YPD fermentation broth, C 1 For the YPD medium not inoculated with Kluyveromyces marxe C2, put the above solution into a 37-degree shaker, shake it for 60 hours, break it up with a homogenizer, and extract it with methanol to be tested. Detection method: Immunoaffinity chromatography purification high performance liquid chromatography, the results are shown in Table 1, Kluyveromyces marxe C2 can grow in the medium containing aflatoxin B1, and can metabolize aflatoxin B1.

[0048] Table 1 Degradation rate of C2 bacteria to aflatoxin B1

[0049] sample

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Abstract

The invention discloses a Kluyveromyces marxianus C2 and a preparation method and an application thereof. The preservation number of the Kluyveromyces marxianus C2 is CCTCC NO: M2014059. The aflatoxin B1, zearalenone and ochratoxin A can be degraded by Kluyveromyces marxianus C2 and the biodegradation rate reaches more than 80%. The fermented Kluyveromyces marxianus C2 is put into toxin-contaminated liquid or solid food or feed matrix at a certain ratio to enable mycotoxins to be fast, safely and efficiently degraded and detoxified and can also be used for oral detoxification of animals. The strain disclosed by the invention has the advantages of low production cost and safe application and has wide application prospects.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to Kluyveromyces marxe C2, which can degrade various mycotoxins, and also relates to a preparation method and application of the yeast. Background technique [0002] According to the estimates of the United Nations Food and Agriculture Organization (FAO), 25% of the grains in the world are polluted by mycotoxins every year, and an average of 2% are inedible; in addition, the diseases and deaths caused by animal poisoning caused by toxin pollution have caused great harm to the grain industry and animal husbandry. Huge economic loss. The survey found that it is also very common that animal raw fishmeal is contaminated with multiple mycotoxins at the same time. After animals eat moldy feed, it will cause weight loss or cause disease. After moldy feed is eaten by animals, toxins and their metabolites may remain in livestock products and their processed products, and enter the ...

Claims

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Application Information

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IPC IPC(8): C12N1/16A23K1/16A23L1/015A61K36/06A61P39/02C12R1/645A23K10/18A23L5/20A23L31/10
Inventor 徐高原王杨波周明光金建云陈章表李芳韩进蔡皓金梅林陈焕春
Owner WUHAN KEQIAN BIOLOGY CO LTD
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