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- W2292190239 abstract "The wide use of pesticides is a threat to the public health. An attractive tool for bioremediation of pesticides pollution is the bacterial pesticides degrading enzyme, phosphotriesterase (OPH), which is able to hydrolyze a wide range of organophosphate compounds. Yet, engineering of a more potent hydrolytic machinery into this enzyme could yield an advantageous bioremediation tool. We undertook the in-vitro evolution strategy in order to improve the hydrolyzing power of phosphotriesterase towards certain organophosphates. The opd gene, coding for the phosphotriesterase enzyme, was amplified from Flavobacterium by PCR, and introduced into pmalE gene fusion system, expressed in E. coli, to allow simple procedure for purification of the recombinant mature native (and future mutant) enzyme. Random mutagenesis of the opd gene was carried out by error-prone PCR, and controlled low level mutagenesis was adopted. Mutagenesis products served for generation of an expression library, using a modified pUC 18 as an expression vector. A screening system was developed to allow monitoring of enzymatic kinetics with whole expressing cells, and to isolate phosphotriesterase mutants with altered enzymatic properties." @default.
- W2292190239 created "2016-06-24" @default.
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- W2292190239 date "1999-01-01" @default.
- W2292190239 modified "2023-09-27" @default.
- W2292190239 title "DIRECTED EVOLUTION OF ABACTERIAL PESTICIDES DEGRADING ENZYME" @default.
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