Preparation method of compound enzyme preparation for efficiently degrading aflatoxin B1

A compound enzyme preparation, aflatoxin technology, applied in the fields of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of little research, achieve significant material saving, high efficiency, and improve the effect of degradability

Inactive Publication Date: 2017-05-10
HENAN UNIVERSITY OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are very few studies on the efficient degradation of aflatoxin B1 by mixing different bacterial extracellular enzymes according to a certain percentage of components to make a compound enzyme preparation

Method used

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  • Preparation method of compound enzyme preparation for efficiently degrading aflatoxin B1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Fermentation and culture of Neurospora palpus and Penicillium raceae, and centrifugation of the respective culture solutions in centrifuge tubes at 5500rpm for 10 minutes, separating the supernatant from the lower layer of bacteria, concentrating the supernatant by ultrafiltration 10 times, using 50 % saturated ammonium sulfate precipitation, centrifuged at 10,000 rpm for 5 minutes, discarded the supernatant, redissolved the precipitated protein in phosphate buffer, dialyzed overnight and then freeze-dried, and then used the obtained protein as the crude enzyme preparation of N. Penicillium penicillium crude enzyme preparation, save for future use.

[0029]Mix the crude enzyme preparation of Neurospora aerugaus, the crude enzyme preparation of Penicillium raceae, and the fungal laccase respectively in component percentages (20%, 20%, 60%), and weigh 0.2g of the compound enzyme preparation (which contains 0.04g Crude enzyme preparation from Neurospora californica, 0.04g ...

Embodiment 2

[0032] On the basis of the preparation of the crude enzyme preparations of Neuromonas aeruginosa and the crude enzyme preparations of Penicillium raceae in Example 1, the crude enzyme preparations of Neuromonas arborescens, the crude enzyme preparations of Penicillium raceus and fungal laccase were respectively divided into components Percentages (40%, 40%, 20%) were mixed to make a compound enzyme preparation, and weighed 0.4g compound enzyme preparation (including 0.16g of N. fungal laccase), fully shaken and dissolved in 1L sterile distilled water to make an enzyme solution, so that the concentration in the system is 0.4g / L, adding aflatoxin B1 standard product to make the initial concentration of aflatoxin B1 in the enzyme solution is 1ppm. At this time, the ratio of the compound enzyme preparation to the aflatoxin B1 in the degradation system was 0.4 g / L·ppm. Control the culture temperature at 36°C, pH at 6.5, and detect the degradation time required when the remaining c...

Embodiment 3

[0035] On the basis of the preparation of the crude enzyme preparations of Neuromonas aeruginosa and the crude enzyme preparations of Penicillium raceae in Example 1, the crude enzyme preparations of Neuromonas arborescens, the crude enzyme preparations of Penicillium raceus and fungal laccase were respectively divided into components Percentage (40%, 40%, 20%), weigh 0.4g compound enzyme preparation (including 0.16g crude enzyme preparation of Neuromonas aerulus, 0.16g crude enzyme preparation of Penicillium laceii, 0.08g fungal laccase), Dissolve in 1L sterile distilled water with sufficient shaking to make an enzyme solution, so that the concentration in the system is 0.4g / L, and add aflatoxin B1 standard substance to make the initial concentration of aflatoxin B1 in the enzyme solution 1ppm. The content of the initial aflatoxin B1 was detected as a control. At this time, the ratio of the compound enzyme preparation to the aflatoxin B1 in the degradation system was 0.4 g / L·...

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Abstract

Crude enzyme preparations are respectively obtained by a method for centrifugation, concentration, ammonium sulfate precipitation, freezing and drying of fermenting medium solutions for bacillus subtilis and penicillium raistrickii, and are mixed with fungal laccase through a specific component ratio, so that a compound enzyme preparation for efficiently degrading aflatoxin B1 is prepared. The bacillus subtilis crude enzyme preparation and the penicillium raistrickii crude enzyme preparation are mixed with the fungal laccase, the respective component ratio occupied is controlled to be 20-60 percent, and the sum is 1. The prepared compound enzyme preparation can efficiently degrade the aflatoxin B1, and the degrading interval is 76-89 percent. Under the same treatment conditions, the degrading efficiency of the compound enzyme preparation is improved by 25 percent or above as compared with that of each of the three components for the aflatoxin B1. When the ratio of the compound enzyme preparation to the aflatoxin B1 in a system is controlled to be 0.2-0.4 g / L.ppm, compared with any component in the compound enzyme preparation, the compound enzyme preparation shows significant material-saving property.

Description

technical field [0001] The invention belongs to the field of enzyme preparations, and relates to a preparation method of a compound enzyme preparation for efficiently degrading aflatoxin B1. Background technique [0002] Aflatoxin is a typical fungal secondary metabolite, which has strong carcinogenic, teratogenic and mutagenic effects. The main organ of Aspergillus flavus is the liver, and can induce liver cell canceration, which brings great harm to humans and animals. The scope of aflatoxin contamination is relatively wide, such as peanuts, corn, dried fruits, milk and other daily foods, because of its huge toxicity, it has attracted more and more attention. The pollution of food by aflatoxin has seriously affected the import and export trade of various countries and brought huge losses to the economy. [0003] Aflatoxin is structurally composed of a difuran ring and an oxanaphthone ring. The unsaturated lactone ring structure and difuran ring in its structural formula ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02A23L5/20
CPCC12N9/0004C12N9/0061C12Y110/03002
Inventor 王乐黄巍殷海成王金水胡元森
Owner HENAN UNIVERSITY OF TECHNOLOGY
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