Fumonisin degrading enzyme, coding gene, recombinant vector, cell, additive and application thereof
A fumonisin and recombinant cell technology, which is applied in the field of biotechnology related to Ming Dynasty, can solve the problems of unutilization, long degradation time, and low removal efficiency, and achieve low pH requirements, environmental friendliness, and good stability.
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Embodiment 1
[0069] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.
[0070] (1) Acquisition of genes
[0071] The following nucleotide fragments were synthesized by artificial chemical synthesis (commissioned by Bao Bioengineering (Dalian) Co., Ltd., the same below); the initiation codon ATG was added to the 5' end of the nucleotide sequence shown in SEQ ID NO:1 Add the Xho I restriction site before the Nde I restriction site and the 3' end codon TAG.
[0072] (2) Construction of recombinant plasmids
[0073] 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:
[0074]
[0075] After agarose gel electrophoresis, the digested product was purified and recovered. Then, ...
Embodiment 2
[0086] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.
[0087] The degrading enzyme was prepared according to the method of Example 1, and the influence of temperature and pH on the enzyme activity was determined, except that SEQ ID NO: 3 was used instead of SEQ ID NO: 1.
Embodiment 3
[0089] This example is used to illustrate the degrading enzyme provided by the present invention and its preparation method and application.
[0090] The degrading enzyme was prepared according to the method of Example 1, and the influence of temperature and pH on the enzyme activity was determined, except that SEQ ID NO: 4 was used instead of SEQ ID NO: 1.
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