title: TBADH activity in water-miscible organic solvents: correlations between enzyme performance, enantioselectivity and protein structure through spectroscopic studies.
abstract: The enantioselective reduction of 2-pentanone to (R)- and (S)-2-pentanol by Thermoanaerobacter (formerly Thermoanaerobium) brockii alcohol dehydrogenase (TBADH) in mixtures of water and water-miscible organic solvents was investigated. Significant enzymatic activity was retained in up to 87% methanol, ethanol and acetonitrile. The changes in enzyme activity as a function of organic solvent were correlated to structural alterations of TBADH with a series of spectroscopic studies (fluorescence, fluorescence quenching and circular dichroism (CD)). Interestingly, this study shows that the tetrameric form of TBADH is not critical for catalytic performance.
authors.affiliation: Bioorganic & Biophysical Chemistry Laboratory, Department of Chemistry & Biomedical Sciences, University of Kalmar, SE-391 82, Kalmar, Sweden. linus.olofsson@hik.se
authors.forename: Linus
authors.initials: L
authors.lastname: Olofsson
authors.forename: Ian A
authors.initials: IA
authors.lastname: Nicholls
authors.forename: Susanne
authors.initials: S
authors.lastname: Wikman
mesh_terms: D000097:Acetonitriles / Q000737:chemistry; D020106:Acrylamide / Q000737:chemistry; D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000302:isolation & purification; D002942:Circular Dichroism; D000431:Ethanol / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D010084:Oxidation-Reduction; D010422:Pentanones / Q000737:chemistry; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013237:Stereoisomerism; D046968:Thermoanaerobacter / Q000201:enzymology*; D014364:Tryptophan / Q000737:chemistry; D014867:Water / Q000737:chemistry*
chemical_list: D000097:Acetonitriles; D010422:Pentanones; D021122:Protein Subunits; D014867:Water; D020106:Acrylamide; D000431:Ethanol; D014364:Tryptophan; D000426:Alcohol Dehydrogenase; C076402:2-pentanone; D000432:Methanol; C032159:acetonitrile