Composite enzyme and additive, their application and method for removing mycotoxin
A technology of mycotoxins and compound enzymes, which is applied in the field of removing mycotoxins and compound enzymes, and can solve problems such as difficult to achieve high-efficiency removal, difficult to remove multiple toxins, and decreased removal efficiency
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[0031] The present invention also provides a preparation method of the above-mentioned enzyme compound preparation, the method comprising: uniformly mixing each component according to the aforementioned weight ratio.
[0032] According to the present invention, the fumonisins may include their respective toxin types, for example, FA1, FA2, FB1, FB2, FB3, FB4, FC1, FC2, FC3, FC4 and FP1, among which fumonisin FB1 is most preferred. The ochratoxins may also include their respective toxin types, for example, ochratoxin A and ochratoxin B.
[0033] In the second aspect, the present invention also discloses an additive, wherein the additive contains the complex enzyme as described above.
[0034] In a preferred situation, the additive uses the complex enzyme provided by the invention as an active ingredient. In the additive, the content of the complex enzyme is 0.001-10 g / kg, preferably 0.01-8 g / kg, more preferably 0.1-5 g / kg. In addition, the additives may also contain solvents ...
preparation example 1
[0056] This preparation example is used to illustrate the preparation of amidase provided by the invention
[0057] (1) Acquisition of genes
[0058] The following nucleotide fragments were synthesized by artificial chemical synthesis (commissioned Bao Biological Engineering (Dalian) Co., Ltd., the same below); after the initiation codon ATG at the 5' end of the nucleotide sequence shown in SEQ ID NO:1 Add an Nde I restriction site, add an Xho I restriction site and a stop codon TAG at the 3' end.
[0059] (2) Construction of recombinant plasmids
[0060] Use restriction endonucleases Nde I and Xho I (purchased from NEB Company) to perform double enzyme digestion on the PET30a plasmid (with His tag, purchased from Invitrogen Company, USA), and digest it in a water bath at 37°C for 4 h, and the enzyme digestion system (50 μL) as follows:
[0061]
[0062] After agarose gel electrophoresis, the digested product was purified and recovered. Then, a ligation reaction was per...
preparation example 2-4
[0071] Preparation example 2-4 is used to illustrate the preparation of amidase provided by the invention
[0072] According to the method of Preparation Example 1, amidases X2-X4 were prepared respectively, the difference is that Preparation Examples 2-4 respectively used SEQ ID NO: 3, SEQ ID NO: 4 and SEQ ID NO: 5 to replace SEQ ID NO: 1 .
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