Immobilized organic phosphorus pesticide degrading enzyme as well as preparation method and application thereof

A technology for organophosphorus pesticides and degrading enzymes, applied in the field of fusion enzymes of SBD mutants and organophosphorus pesticide degrading enzymes, immobilized fusion enzymes, starch binding domain mutants, and treatment of organophosphorus pesticide production wastewater

Active Publication Date: 2016-01-06
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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  • Immobilized organic phosphorus pesticide degrading enzyme as well as preparation method and application thereof
  • Immobilized organic phosphorus pesticide degrading enzyme as well as preparation method and application thereof
  • Immobilized organic phosphorus pesticide degrading enzyme as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

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 code word of the starch binding domain (SBD) is optimized according to Pichia pastoris (SEQ ID No.1), which The encoded amino acid sequence is shown in SEQ ID No.2. Respectively add restriction endonuclease XhoI restriction endonuclease restriction endonuclease XhoI restriction endonuclease site at its front end, back end add restriction endonuclease KpnI restriction endonuclease restriction endonuclease restriction site, and join peptide Linker at the front end of KpnI restriction endonuclease restriction site as shown in SEQIDNo.8 , to maintain flexibility between SBD and other fusion enzymes. The connecting peptide comes from the amino aci...

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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 invention relates to the field of biological technology, and specifically relates 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 treating organic Method for phosphorus 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), due to the advantages of high drug efficacy, wide control range, and low cost, are the most widely used pesticides in agricultural production (SinghBK.2009. Organophosphorus-degradingbacteria: ecology and industrial applications. Nature Reviews Microbiology. 164). However, with the development of agriculture, the use of organophosphorus pesticides is increasing, and a large amount of organophosphorus pesticides...

Claims

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Application Information

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Patent Type & Authority Applications(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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