Argüello, José M.
Nickel acquisition is important for many biological and biochemical activities, such as urease activity. A 621AA long hypothetical heavy-metal transporting ATPase (Swiss-port accession #Q9Y8R2, gene name APE 2571) of Aeropyrum pernix was identified as a Nickel/Cadmium/Cobalt transporter CPx ATPase and named CotA. APE2571 was cloned and expressed in Escherichia coli. The expression was checked by immunoblotting after sodium dodecyl sulfate-polyacrylamide gel electrophoresis with an anti-His antibody. CotA enzymatic activity and Ni? binding affinity were studied by ATPase assay. Three methods of bacteria cell lysis, French press, beads beater and sonication were tested for optimum yield of CopA and its specific activity. Beads beater was determined having advantage on time saving and preserving CopA's activity.
Worcester Polytechnic Institute
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Chemistry and Biochemistry