A kind of immobilized organophosphorus pesticide degrading enzyme and its preparation method and application

A technology for organophosphorus pesticides, degrading enzymes, applied in the directions of immobilization on/in organic carriers, biochemical equipment and methods, chemical instruments and methods, etc.

Active Publication Date: 2018-11-02
冯鹏波
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there have been no reports on the use of wild-type SBD or its mutants in the isolation and purification of organophosphorus pesticide degrading enzymes, let alone the use of corn-immobilized organophosphorus pesticide degrading enzymes in the treatment of pesticide production wastewater

Method used

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  • A kind of immobilized organophosphorus pesticide degrading enzyme and its preparation method and application
  • A kind of immobilized organophosphorus pesticide degrading enzyme and its preparation method and application
  • A kind of immobilized organophosphorus pesticide degrading enzyme and its preparation method and application

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Experimental program
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Embodiment Construction

[0061] 1. Wild-type starch binding domain (SBD-connecting peptide (Linker), organophosphate pesticide degrading enzyme OP M164F DNA Design and Synthesis

[0062] According to the nucleotide sequence of Rhizopus oryzae glucoamylase with sequence number ABI75077.1 recorded in the global public sequence database GeneBank as a template, the codon of the starch binding domain (SBD) was optimized according to Pichia pastoris (SEQ ID No.1) , the encoded amino acid sequence is shown in SEQ ID No.2. Respectively add restriction endonuclease XhoI restriction endonuclease site at its front end, rear end add restriction endonuclease KpnI restriction endonuclease restriction site, and join peptide Linker such as SEQ ID No.8 at the front end of KpnI restriction endonuclease site shown to maintain flexibility between SBD and other fusion enzymes. The connecting peptide is derived from the amino acid sequence between domains I and II of human serum albumin, its codons have been optimized ac...

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Abstract

The invention provides a starch binding domain mutant of rhizopus oryzae glucoamylase, a fusion enzyme of the mutant and an organic phosphorus pesticide degrading enzyme, a preparation method and an application in treating pesticide waste water. By analyzing a great amount of binding sites of a starch binding domain, a plurality of binding site mutants are acquired, and a result shows that the mutant SBDm (S33T, F58W) has excellent performance that the mutant can be used as an affinity tag to be bonded with a starch matrix. The mutant SBDm is fused with the organic phosphorus pesticide degrading enzyme (OP), the organic phosphorus pesticide degrading enzyme can be rapidly immobilized on the starch matrix from fermenting liquid in one step, and the enzyme activity is not affected by the affinity tag and immobilization. The degrading enzyme after being immobilized on the starch matrix has a vast application prospect on the aspects of treating the organic phosphorus pesticide production waste water and removing the residue of other pesticides.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a starch-binding domain (SBD) mutant, a fusion enzyme of an SBD mutant and an organophosphorus pesticide degrading enzyme, an immobilized fusion enzyme, and the use of the immobilized enzyme for processing A method for organophosphorus pesticide production wastewater. Background technique [0002] Pesticides have made great contributions to the development of modern agriculture and human food supply. Organophosphorus pesticides (organophosphorus pesticides) are the most widely used pesticides in agricultural production due to their high efficacy, wide range of control and low cost (Singh BK. 2009. Organophosphorus-degrading bacteria: ecology and industrial applications. Nature Reviews Microbiology. 7:156-164). However, with the development of agriculture, the use of organophosphorus pesticides is increasing, and a large amount of organophosphorus pesticides remain in water ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/34C12N15/56C12N9/16C12N9/80C12N9/88C12N15/81C12N11/10C02F3/34C02F101/30
CPCC02F3/342C02F2101/306C07K2319/20C12N9/16C12N9/2428C12N9/80C12N9/88C12N11/10C12N15/62C12Y302/01003
Inventor 徐建华陈江泽卡拉·然·徐冯鹏波
Owner 冯鹏波
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