caa a22 4500 397556438 CHVBK 20180308164803.0 cr unu---uuuuu 161202e199609 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021441 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021441 A Thermophilic Alcohol Dehydrogenase from Bacillus acidocaldarius Not Reactive towards Ketones [Elektronische Daten] [Sabato D'Auria, Francesco La Cara, Filomena Nazzaro, Nunzia Vespa, Mose Rossi] An NAD-dependent alcohol-aldehyde oxidoreductase was purified to homogeneity and characterized from cell extracts of the thermophilic microorganism Bacillus acidocaldarius. The 500-fold purified homogeneous enzyme had a molecular mass of 154 kDa, as shown by gel filtration and glycerol gradient centrifugation. On sodium dodecyl sulfate polyacrylamide gel electrophoresis the protein showed one band of 38 kDa, indicating that the enzyme is a tetramer composed of subunits of identical molecular weight. Ethanol was the best substrate with the highest kcat/Km values, and the enzyme showed a substrate specificity that included linear, secondary and cyclic alcohols, as well as anisaldehyde, but it was not active on ketones. The protein contains eight zinc atoms per tetrainer, four of which are removed by chelating agents with a concomitant loss of thermal stability. Circular dichroism spectra and determination of the NH2-terminal sequence allowed structural and homology comparison with other alcohol dehydrogenases from animal and bacterial sources. © 1996 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence alcohol dehydrogenase nationallicence Bacillus acidocaldarius nationallicence circular dichroism nationallicence en-zymatic activity nationallicence thermophilic enzymes nationallicence D'Auria Sabato Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut La Cara Francesco Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut Nazzaro Filomena Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut Vespa Nunzia Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut Rossi Mose Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut The Journal of Biochemistry Oxford University Press 120/3(1996-09), 498-504 0021-924X 120:3<498 1996 120 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021441 text/html Onlinezugriff via DOI 1 other jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021441 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- D'Auria Sabato Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut NATIONALLICENCE 700 1- La Cara Francesco Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut NATIONALLICENCE 700 1- Nazzaro Filomena Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut NATIONALLICENCE 700 1- Vespa Nunzia Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut NATIONALLICENCE 700 1- Rossi Mose Institute of Protein Biochemistry and Enzymology C.N.R. Via Marconi, 10, 80125 Naples, Italy aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 120/3(1996-09), 498-504 0021-924X 120:3<498 1996 120 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford