Characterization of Bacillus anthracis arginase: effects of pH, temperature, and cell viability on metal preference.
BACKGROUND
Arginase (RocF) hydrolyzes L-arginine to L-ornithine and urea. While previously characterized arginases have an alkaline pH optimum and require activation with manganese, arginase from Helicobacter pylori is optimally active with cobalt at pH 6. The arginase from Bacillus anthracis is not well characterized; therefore, this arginase was investigated by a variety of strategies and the enzyme was purified.


RESULTS
The rocF gene from B. anthracis was cloned and expressed in E. coli and compared with E. coli expressing H. pylori rocF. In the native organisms B. anthracis arginase was up to 1,000 times more active than H. pylori arginase and displayed remarkable activity in the absence of exogenous metals, although manganese, cobalt, and nickel all improved activity. Optimal B. anthracis arginase activity occurred with nickel at an alkaline pH. Either B. anthracis arginase expressed in E. coli or purified B. anthracis RocF showed similar findings. The B. anthracis arginase expressed in E. coli shifted its metal preference from Ni > Co > Mn when assayed at pH 6 to Ni > Mn > Co at pH 9. Using a viable cell arginase assay, B. anthracis arginase increased dramatically when the cells were grown with manganese, even at final concentrations of <1 muM, whereas B. anthracis grown with cobalt or nickel (> or =500 microM) showed no such increase, suggesting existence of a high affinity and specificity manganese transporter.


CONCLUSIONS
Unlike other eubacterial arginases, B. anthracis arginase displays unusual metal promiscuity. The unique properties of B. anthracis arginase may allow utilization of a specific metal, depending on the in vivo niches occupied by this organism.
Department of Biology, University of South Alabama, Mobile, AL 36688, USA. rjv302@jaguar1.usouthal.edu
Ryan J
RJ
Viator
Richard F
RF
Rest
Ellen
E
Hildebrandt
David J
DJ
McGee
D001119:Arginase / Q000378:metabolism*; D001408:Bacillus anthracis / Q000201:enzymology* / Q000254:growth & development / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002470:Cell Survival / Q000502:physiology*; D003035:Cobalt / Q000737:chemistry / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008345:Manganese / Q000737:chemistry / Q000378:metabolism; D019216:Metals, Heavy / Q000737:chemistry* / Q000378:metabolism; D009532:Nickel / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
D001426:Bacterial Proteins; D019216:Metals, Heavy; D003035:Cobalt; D008345:Manganese; D009532:Nickel; D001119:Arginase