Abstract
Supernatant fluid from Leptothrix discophora SS-1 cultures possessed high Mn2+-ozidizing activity. Studies of temperature and pH optima, chemical inhibition, and protease sensitivity suggested that the activity may be enzymatic. Kinetic studies of unconcentrated supernatant fluid indicated an apparent Km of 7 microM Mn2+ in the 1 to 200 microM Mn2+ range. The greatest Vmax value observed was 1.4 nmol of Mn2+ oxidized min-1 micrograms of protein-1 in unconcentrated samples. When the supernatant fluid was concentrated on DEAE-cellulose and the activity was eluted with MgSO4, an Mn2+-oxidizing protein was detected in the concentrate by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Mn2+-oxidizing protein appeared to have a molecular weight of 110,000 in 10% polyacrylamide gels and of 100,000 in 8% gels. Periodic acid-Schiff base staining of overloaded polyacrylamide gels showed that the DEAE-cellulose concentrate contained abundant high-molecular-weight polysaccharides; concurrent staining of the Mn2+-oxidizing band suggested that it too contained carbohydrate components. Isolation of the protein was achieved by subjecting the DEAE-cellulose concentrate to Sephacryl gel filtration in the presence of 1% sodium dodecyl sulfate, followed by preparative electrophoresis and reverse-polarity elution. However, these procedures resulted in loss of a large proportion of the activity, which precluded recovery of the protein in significant quality.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Reference19 articles.
1. Recovery of native proteins from preparative electrophoresis gel slices by reverse polarity elution;Abramovitz A. S.;Prep. Biochem.,1984
2. Influence of manganese on growth of a sheathless strain of Leptothrix discophora;Adams L. F.;Appl. Environ. Microbiol.,1985
3. Physiology and ultrastructure of Leptothrix discophora SS-1;Adams L. F.;Arch. Microbiol.,1986
4. Stimulation of heterotrophic and autotrophic growth of Sphaerotilus discophorus by manganous ions. Antonie van Leeuwenhoek J;Ali S. H.;Microbiol. Serol.,1971
5. Manganese oxidation by Leptothrix discophora;Boogerd F. C.;J. Bacteriol.,1986
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