Surface Display of Methyl Parathion Hydrolase on Saccharomyces cerevisiae and Its Application in Degradation of Methyl Parathion

doi:10.13560/j.cnki.biotech.bull.1985.2015.04.026

Abstract

Abstract: The methyl parathion hydrolase(MPH)gene of Pseudomonas sp.WBC-3 was amplified by PCR and cloned into the multiple cloning site of the surface display vector pYD1 to construct a recombinant plasmid pYD1-MPH. Then plasmid pYD1-MPH was transformed into Saccharomyces cerevisiae EBY100. The 2% galactose was used to induce the expression of MPH on the cell surface of EBYl00, and the display of MPH on the cell surface of S.cerevisiae was confirmed by immunofluorescence. The characteristic of the displayed MPH and degradation effect of methyl parathion in water by the engineered yeast were also investigated. The result showed that the engineered yeast strain, which have a whole cell catalytic activity of MPH, was successfully constructed. The activity of MPH displayed on the yeast cells was 18.2 U/mg of cell dry cells by the induction of 2% galactose for 48 h. The displayed MPH had the optimal pH of 9.5 and the optimal temperature of 30℃, respectively and was stable in the pH range of 4.0-10.5 and up to 45℃. The displayed MPH was stimulated by Mn²⁺，Co²⁺，Zn²⁺，Ca²⁺，Hg²⁺，K⁺，Ni²⁺, and was inhibited by Na⁺，Fe³⁺，Ag⁺. The engineered yeast strain could hydrolyze over 80% of 20.0 mg/L methyl parathion in tap water in 1 h.

Key words: methyl parathion hydrolase, surface display, Saccharomyces cerevisiae, methyl parathion degradation

Wang Xingxing, Chi Zhe. Surface Display of Methyl Parathion Hydrolase on Saccharomyces cerevisiae and Its Application in Degradation of Methyl Parathion[J]. Biotechnology Bulletin, 2015, 31(3): 178-184.

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