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Zearalenone degrading enzyme fusion protein and coding gene thereof and application thereof

A technology of zearalenone and fusion protein, applied in the field of agricultural biology, can solve the problems of low expression amount, high cost, time-consuming and the like, and achieve the effects of increasing expression amount and improving activity

Active Publication Date: 2019-07-26
INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Gene zhd101 has been confirmed to have the ability to degrade zearalenone, but its expression level in exogenous expression system is low. ), thin-layer chromatography (TCL method), immunoaffinity column-high performance liquid chromatography (IAC-HPLC), enzyme-linked immunosorbent assay (ELISA), etc., the operation is more complicated, time-consuming and costly

Method used

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  • Zearalenone degrading enzyme fusion protein and coding gene thereof and application thereof
  • Zearalenone degrading enzyme fusion protein and coding gene thereof and application thereof
  • Zearalenone degrading enzyme fusion protein and coding gene thereof and application thereof

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Embodiment 1

[0046] 1. Amplify the fusion protein sequence

[0047] PCR method was used to amplify the zhd101 and xynb gene fragments, and the recombinant plasmid pPICZ(α)A-xynb-linker-zhd101 was obtained by connecting with the vector pPICZ(α)A through the recombination kit, and transformed into Pichia pastoris X33 to obtain the recombinant Pichia red yeast strain X33 / xynb-linker-zhd101.

[0048] The primers used in PCR are as follows:

[0049] xynb-pPICZ(α)A:

[0050] F: 5'-AGAGAGGCTGAAGCTGAATTCGTTCCACACGACTCTGTTGCTCAAAGATCC

[0051] R: 5'-TGGTGGTTGGAGTAGTACCACCGGAAGAAGCAGATTGAACAGTAATGGAGGAGGA

[0052] zhd101-pPICZ(α)A

[0053] F: 5'-GCTTCTTCCGGTGGTACTACTCCAACCACCACTATGAGAACTAGATCTACTATT

[0054] R: 5'-TTCTAGAAAGCTGGCGGCCGCTTACAAATATTTTTTGAGTAGTCTCAACAAC

[0055] pPICZ(α)A-xynb-linker-zhd101

[0056] F: 5'-GAGACTACTCAAAAAATATTTGTAAGCGGCCGCCAGCTTTCTAGAACAAAAA

[0057] R: 5'-AGCAACAGAGTCGTGTGGAACGAATTCAGCTTCAGCCTCTCTTTTTCTCGAG

[0058] Among them, xynb-pPICZ(α)AF and xynb-pPICZ(α)...

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Abstract

The invention belongs to the field of agricultural biotechnology, and particularly relates to a zearalenone degrading enzyme fusion protein and a coding gene thereof and application thereof. The zearalenone degrading enzyme fusion protein is composed of a zearalenone degrading enzyme ZHD101 and xylanase XYNB, and an amino acid sequence of the zearalenone degrading enzyme ZHD101 is as shown in SEQID No. 1, and an amino acid sequence of the xylanase XYNB is as shown in SEQ ID No. 2. According to the zearalenone degrading enzyme fusion protein and the coding gene thereof and the application thereof, the expression level of the zearalenone degrading enzyme is greatly increased, and the enzyme activity is 2.2 times that of ZHD101 expressed alone.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a zearalenone-degrading enzyme fusion protein and its coding gene and application. Background technique [0002] Zearalenone (Zearalenone, ZEN), also known as F-2 toxin, its chemical name is 6-(10-hydroxy-6-oxyl-undecenyl) β-rapenolide, which is a It is a secondary metabolite mainly produced by fungi such as Fusarium graminearum, Fusarum culmorum and Fusarum crookwellense, and it is also the most widely polluted Fusarium in the world toxin. The molecular formula of ZEN is C 18 h 23 O, the average molecular weight is 318Da, white crystal, the boiling point is 510.12°C, the melting point is 217.32°C, relatively stable to heat, it will not be degraded when heated at 120°C for 4 hours. ZEN is insoluble in water, soluble in alkaline aqueous solution, ether, benzene, chloroform, alcohols and acids, slightly soluble in petroleum ether. [0003] At present, the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/81C12N1/19A23L5/20C12R1/84
CPCA23L5/27C12N9/16C12N9/248C12N15/815
Inventor 姚斌柏映国罗会颖于心蕊陈家明涂涛孟昆王亚茹黄火清苏小运王苑师霞
Owner INST OF ANIMAL SCI OF CHINESE ACAD OF AGRI SCI
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