Enhanced tolerance and accumulation of heavy metal ions by engineered Escherichia coli expressing Pyrus calleryana phytochelatin synthase.
Contamination by heavy metals is a major environmental problem worldwide and microbial bioremediation is an efficient method for removing this type of pollution. The plant enzymephytochelatin synthase (PCS, also known as glutathione g-glutamylcysteinyltransferase, EC2.3.2.15) involved in the synthesis of phytochelatins (PCs), which are metal-binding cysteine-rich peptides, has a major role in the detoxification of heavy metals in plants. Expression of the PcPCS1 gene from the bean pear (Pyrus calleryana Dcne.) was induced after cadmium and copper treatments. However, functional analysis of this gene in vivo has not been reported. And it is or not suitable for bioremediation also needs to be assessed. In this study, we found Escherichia coli with over-expressed PcPCS1 had enhanced tolerance to cadmium, copper, sodium, and mercury. E. colicells transformed with pPcPCS1 was found to survive in solid M9 medium containing 2.0 mM Cd(2+), 4.0 mM Cu(2+). 4.5% (w/v) Na+, or 200 μ MHg(2+). Moreover, the growth curve showed 1.5 mM Cd(2+), 2.5 mM Cu(2+), 3.5% (w/v) Naþ, and 100 μ MHg(2+) had no effect on the growth of the E. coli cells transformed with pPcPCS1. Also, we found the contents of PCs and the accumulation of cadmium,copper, sodium, and mercury ions were enhanced in the recombinant E. coli strain Rosetta(TM) (DE3).These results suggested the PcPCS1 gene might be a candidate for heavy metal bioremediation via recombinant bacteria.
Institute of Horticulture, Jiangsu Academy of Agricultural Sciences, Nanjing, P. R. China.
Hui
H
Li
Yu
Y
Cong
Jing
J
Lin
Youhong
Y
Chang
D019881:Aminoacyltransferases / Q000235:genetics* / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002104:Cadmium / Q000378:metabolism; D002681:China; D003300:Copper / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*; D008628:Mercury / Q000378:metabolism; D019216:Metals, Heavy / Q000378:metabolism*; D054811:Phytochelatins / Q000378:metabolism; D031989:Pyrus / Q000201:enzymology / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012964:Sodium / Q000378:metabolism
D019216:Metals, Heavy; D011993:Recombinant Fusion Proteins; D002104:Cadmium; D003300:Copper; D054811:Phytochelatins; D012964:Sodium; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase; D008628:Mercury