Direct identification of a bacterial manganese(II) oxidase, the multicopper oxidase MnxG, from spores of several different marine Bacillus species.
Microorganisms catalyze the formation of naturally occurring Mn oxides, but little is known about the biochemical mechanisms of this important biogeochemical process. We used tandem mass spectrometry to directly analyze the Mn(II)-oxidizing enzyme from marine Bacillus spores, identified as an Mn oxide band with an in-gel activity assay. Nine distinct peptides recovered from the Mn oxide band of two Bacillus species were unique to the multicopper oxidase MnxG, and one peptide was from the small hydrophobic protein MnxF. No other proteins were detected in the Mn oxide band, indicating that MnxG (or a MnxF/G complex) directly catalyzes biogenic Mn oxide formation. The Mn(II) oxidase was partially purified and found to be resistant to many proteases and active even at high concentrations of sodium dodecyl sulfate. Comparative analysis of the genes involved in Mn(II) oxidation from three diverse Bacillus species revealed a complement of conserved Cu-binding regions not present in well-characterized multicopper oxidases. Our results provide the first direct identification of a bacterial enzyme that catalyzes Mn(II) oxidation and suggest that MnxG catalyzes two sequential one-electron oxidations from Mn(II) to Mn(III) and from Mn(III) to Mn(IV), a novel type of reaction for a multicopper oxidase.
Dick
Gregory J
GJ
Marine Biology Research Division, Scripps Institution of Oceanography, University of California San Diego, 9500 Gilman Drive, La Jolla, California 92093-02021, USA.
Torpey
Justin W
JW
Beveridge
Terry J
TJ
Tebo
Bradley M
BM
D001407:Bacillus; D001483:Base Sequence; D003300:Copper / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D008345:Manganese / Q000378:metabolism*; D008386:Marine Biology; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013171:Spores, Bacterial / Q000201:enzymology* / Q000648:ultrastructure; D053719:Tandem Mass Spectrometry; D014871:Water Microbiology
D008345:Manganese; D003300:Copper; D010088:Oxidoreductases