A Molecular Biotechnology For Removal of Toxic Heavy Metals

The thiosulfate reductase gene (PhsABC) from Salmonella typhimurium was expressed in Escherichia coli in order to produce sulfide from inorganic thiosulfate and precipitate metals as metal sulfide complexes. A 5.1-kb DNA fragment containing the native phsABC and a 3.7-kb DNA fragment, excluding putative promoter and regulatory regions were inserted into expression vectors pTrc99A and pJB866, respectively. Upon expression of phsABC, E. coli DH5$\alpha$ harboring the phsABC constructs showed higher thiosulfate reductase activity and produced significantly more sulfide than the control strain (E. coli DH5$\alpha$) under both aerobic and anaerobic conditions. Among the four constructs, E. coli DH5$\alpha$ harboring pSB74 produced the highest level of thiosulfate reductase and removed most of heavy metals from solution under anaerobic conditions. In a mixture of 100 $\mu$M each of cadmium, lead, and zinc, the strain could remove $99\%$ of the total metals from solution within 10 hours. Cadmium was removed first, lead second, and zinc last. In contrast, a negative control did not produce any measurable sulfide and removed very little metals from solution. These results have important implications for removal of metals from wastewater contaminated with several metals.