title: Enhanced arsenic accumulation by engineered yeast cells expressing Arabidopsis thaliana phytochelatin synthase.
abstract: Phytochelatins (PCs) are naturally occurring peptides with high-binding capabilities for a wide range of heavy metals including arsenic (As). PCs are enzymatically synthesized by phytochelatin synthases and contain a (gamma-Glu-Cys)(n) moiety terminated by a Gly residue that makes them relatively proteolysis resistant. In this study, PCs were introduced by expressing Arabidopsis thaliana Phytochelatin Synthase (AtPCS) in the yeast Saccharomyces cerevisiae for enhanced As accumulation and removal. PCs production in yeast resulted in six times higher As accumulation as compared to the control strain under a wide range of As concentrations. For the high-arsenic concentration, PCs production led to a substantial decrease in levels of PC precursors such as glutathione (GSH) and gamma-glutamyl cysteine (gamma-EC). The levels of As(III) accumulation were found to be similar between AtPCS-expressing wild type strain and AtPCS-expressing acr3Delta strain lacking the arsenic efflux system, suggesting that the arsenic uptake may become limiting. This is further supported by the roughly 1:3 stoichiometric ratio between arsenic and PC2 (n = 2) level (comparing with a theoretical value of 1:2), indicating an excess availability of PCs inside the cells. However, at lower As(III) concentration, PC production became limiting and an additive effect on arsenic accumulation was observed for strain lacking the efflux system. More importantly, even resting cells expressing AtPCS pre-cultured in Zn(2+) enriched media showed PCs production and two times higher arsenic removal than the control strain. These results open up the possibility of using cells expressing AtPCS as an inexpensive sorbent for the removal of toxic arsenic.
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside, California 92521.
authors.forename: Shailendra
authors.initials: S
authors.lastname: Singh
authors.forename: Wonkyu
authors.initials: W
authors.lastname: Lee
authors.forename: Nancy A
authors.initials: NA
authors.lastname: Dasilva
authors.forename: Ashok
authors.initials: A
authors.lastname: Mulchandani
authors.forename: Wilfred
authors.initials: W
authors.lastname: Chen
mesh_terms: D019881:Aminoacyltransferases / Q000235:genetics / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology*; D001151:Arsenic / Q000302:isolation & purification / Q000378:metabolism*; D001673:Biodegradation, Environmental; D004785:Environmental Pollutants / Q000302:isolation & purification / Q000378:metabolism*; D005818:Genetic Engineering; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics
chemical_list: D004785:Environmental Pollutants; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase; D001151:Arsenic