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Method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungus fermentation liquor

A technology of aflatoxin and fermentation liquid, applied in the direction of enzymes, biochemical equipment and methods, enzymes, etc., to achieve the effects of protecting biological activity, meeting industrial needs, and simplifying operations

Inactive Publication Date: 2021-08-03
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to solve the shortcomings existing in the existing extraction process of aflatoxin degrading mold, and to provide a method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungal fermentation broth. This method utilizes plant-derived tannins to precipitate proteins The characteristics of AFB in the fermentation supernatant of Aspergillus terreus 1 The degrading enzyme and tannin form a complex and precipitate, polyethylene glycol (PEG6000) is easy to combine with the tannin in the precipitation complex, so that the AFB1 degrading enzyme can be resolved, and the AFB1 degrading enzyme can be separated efficiently while preserving the biological activity of the enzyme Purpose of purification

Method used

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  • Method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungus fermentation liquor
  • Method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungus fermentation liquor
  • Method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungus fermentation liquor

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Embodiment 1

[0022] A method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungal fermentation broth, comprising the following steps:

[0023] Step 1: fungal fermentation, select Aspergillus terreus to cultivate in the culture medium, filter the fermentation liquid through medium-speed filter paper, centrifuge and collect supernatant A in a centrifuge tube to obtain fungal fermentation liquid, the preservation number of Aspergillus terreus is CGMCC N0: 18840, Medium conditions of fungal fermentation broth: 3g / L beef extract, 10g / L peptone, 6g / L glucose, 5g / L NaCl, prepared with water, pH 7.0, autoclaved at 121°C, fermentation temperature 25-37°C , Fermentation time 3-7d, shaker speed 150-180rpm, shaker flask type is three-concave triangular flask;

[0024] Step 2: Composite precipitation of AFB1 degrading enzyme, add tannin to the centrifuge tube containing fungal fermentation broth in step 1, wherein the concentration of tannin is: 10mg / mL, after fully mixin...

Embodiment 2

[0028] A method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungal fermentation broth, comprising the following steps:

[0029] Step 1: fungal fermentation, select Aspergillus terreus to cultivate in the culture medium, filter the fermentation liquid through medium-speed filter paper, centrifuge and collect supernatant A in a centrifuge tube to obtain fungal fermentation liquid, the preservation number of Aspergillus terreus is CGMCC N0: 18840, Medium conditions of fungal fermentation broth: 3g / L beef extract, 10g / L peptone, 6g / L glucose, 5g / L NaCl, prepared with water, pH 7.0, autoclaved at 121°C, fermentation temperature 25-37°C , Fermentation time 3-7d, shaker speed 150-180rpm, shaker flask type is three-concave triangular flask;

[0030] Step 2: Composite precipitation of AFB1 degrading enzyme, add tannin to the centrifuge tube containing fungal fermentation broth in step 1, wherein the concentration of tannin is: 40mg / mL, after fully mixin...

Embodiment 3

[0034] A method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungal fermentation broth, comprising the following steps:

[0035]Step 1: fungal fermentation, select Aspergillus terreus to cultivate in the culture medium, filter the fermentation liquid through medium-speed filter paper, centrifuge and collect supernatant A in a centrifuge tube to obtain fungal fermentation liquid, the preservation number of Aspergillus terreus is CGMCC N0: 18840, Medium conditions of fungal fermentation broth: 3g / L beef extract, 10g / L peptone, 6g / L glucose, 5g / L NaCl, prepared with water, pH 7.0, autoclaved at 121°C, fermentation temperature 25-37°C , Fermentation time 3-7d, shaker speed 150-180rpm, shaker flask type is three-concave triangular flask;

[0036] Step 2: Composite precipitation of AFB1 degrading enzyme, add tannin to the centrifuge tube containing fungal fermentation broth in step 1, wherein the concentration of tannin is: 60mg / mL, after fully mixing...

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Abstract

The invention discloses a method for efficiently separating and extracting aflatoxin B1 (AFB1) degrading enzyme from fungus fermentation liquor, and relates to the technical field of biology. The method comprises the steps: step 1, fungus fermentation: selecting aspergillus terreus to be cultured in a culture medium, filtering the fermentation liquor through medium-speed filter paper, carrying out centrifugation, collecting supernate A in a centrifugal tube, and preparing the fungus fermentation liquor; step 2, composite precipitation of the AFB1 degrading enzyme: adding tannin into a centrifuge tube containing the fungal fermentation liquor in the step 1, adding tannin with a concentration of 10-80 mg / mL into each mL of the fungal fermentation liquor, fully and uniformly mixing, precipitating for 10-70 min, centrifuging for 5-20 min at a rotating speed of 5000-10000 rpm, and collecting a composite precipitate C; step 3, redissolving and extracting of the AFB1 degrading enzyme: fully redissolving the composite precipitate C prepared in the step 2 by using a PEG6000 solution with the concentration of 10-40 mg / mL, so as to prepare an AFB1 degrading enzyme solution; and step 4, determining the degradation rate of the AFB1 degrading enzyme liquid, and achieving the purpose of efficient separation and purification of the AFB1 degrading enzyme while preserving the biological activity of the enzyme.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for efficiently separating and extracting aflatoxin B1 degrading enzyme from fungal fermentation broth. Background technique [0002] Since the outbreak of mycotoxin disease in turkeys in the United Kingdom in 1960, a large number of investigations and studies on mycotoxins have been carried out around the world. Currently, more than 300 species of fungi are known to produce mycotoxins. The main mycotoxins include: aflatoxin, zearalenone, ochratoxin, trichothecenes, and citrinin. These toxins are widely distributed and have been isolated from a large number of feed ingredients and mixed feeds. Among various mycotoxins, aflatoxin is considered the most serious and toxic, because it can cause liver poisoning, and has strong immunosuppressive, carcinogenic, mutagenic and teratogenic properties. According to the data of the Food and Agriculture Organization of the United Natio...

Claims

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Application Information

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IPC IPC(8): C12N9/00
CPCC12N9/00
Inventor 谢岩黎王佳兴孙淑敏马卫宾李倩杨玉辉刘晨
Owner HENAN UNIVERSITY OF TECHNOLOGY
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