Nitrate-Dependent Degradation of Acetone by Alicycliphilus and Paracoccus Strains and Comparison of Acetone Carboxylase Enzymes
- 1 October 2011
- journal article
- Published by American Society for Microbiology in Applied and Environmental Microbiology
- Vol. 77 (19), 6821-6825
- https://doi.org/10.1128/aem.05385-11
Abstract
A novel acetone-degrading, nitrate-reducing bacterium, strain KN Bun08, was isolated from an enrichment culture with butanone and nitrate as the sole sources of carbon and energy. The cells were motile short rods, 0.5 to 1 by 1 to 2 μm in size, which gave Gram-positive staining results in the exponential growth phase and Gram-negative staining results in the stationary-growth phase. Based on 16S rRNA gene sequence analysis, the isolate was assigned to the genus Alicycliphilus . Besides butanone and acetone, the strain used numerous fatty acids as substrates. An ATP-dependent acetone-carboxylating enzyme was enriched from cell extracts of this bacterium and of Alicycliphilus denitrificans K601 T by two subsequent DEAE Sepharose column procedures. For comparison, acetone carboxylases were enriched from two additional nitrate-reducing bacterial species, Paracoccus denitrificans and P. pantotrophus . The products of the carboxylase reaction were acetoacetate and AMP rather than ADP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of cell extracts and of the various enzyme preparations revealed bands corresponding to molecular masses of 85, 78, and 20 kDa, suggesting similarities to the acetone carboxylase enzymes described in detail for the aerobic bacterium Xanthobacter autotrophicus strain Py2 (85.3, 78.3, and 19.6 kDa) and the phototrophic bacterium Rhodobacter capsulatus . Protein bands were excised and compared by mass spectrometry with those of acetone carboxylases of aerobic bacteria. The results document the finding that the nitrate-reducing bacteria studied here use acetone-carboxylating enzymes similar to those of aerobic and phototrophic bacteria.Keywords
This publication has 26 references indexed in Scilit:
- ATP-Dependent Carboxylation of Acetophenone by a Novel Type of CarboxylaseJournal of Bacteriology, 2010
- Growth Yields in Bacterial Denitrification and Nitrate AmmonificationApplied and Environmental Microbiology, 2007
- ATP-Dependent Enolization of Acetone by Acetone Carboxylase from Rhodobacter capsulatusBiochemistry, 2005
- Biochemical, Molecular, and Genetic Analyses of the Acetone Carboxylases from Xanthobacter autotrophicus Strain Py2 and Rhodobacter capsulatus Strain B10Journal of Bacteriology, 2002
- 14CO2 Exchange with Acetoacetate Catalyzed by Dialyzed Cell-Free Extracts of the Bacterial Strain Bunn Grown with Acetone and NitrateJBIC Journal of Biological Inorganic Chemistry, 1995
- Anaerobic Degradation of Acetone and Higher Ketones via Carboxylation by Newly Isolated Denitrifying BacteriaMicrobiology, 1989
- Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G‐250 and R‐250Electrophoresis, 1988
- The Microbial Metabolism of AcetoneMicrobiology, 1980
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970
- Variables Influencing Results, and the Precise Definition of Steps in Gram Staining as a Means of Standardizing the Results ObtainedStain Technology, 1962