Insight into the stereospecificity of short-chain thermus thermophilus alcohol dehydrogenase showing pro-S hydride transfer and prelog enantioselectivity.
The stereochemistry of the hydride transfer in reactions catalyzed by NAD(H)-dependent alcohol dehydrogenase from Thermus thermophilus HB27 was determined by means of (1)H-NMR spectroscopy. The enzyme transfers the pro-S hydrogen of [4R-(2)H]NADH and exhibits Prelog specificity. Enzyme-substrate docking calculations provided structural details about the enantioselectivity of this thermophilic enzyme. These results give additional insights into the diverse active site architectures of the largely versatile short-chain dehydrogenase superfamily enzymes. A feasible protocol for the synthesis of [4R-(2)H]NADH with high yield was also set up by enzymatic oxidation of 2-propanol-d(8) catalyzed by Bacillus stearothermophilus alcohol dehydrogenase.
Pennacchio
Angela
A
Istituto di Biochimica delle Proteine, CNR, Via P. Castellino 111, I-80131, Naples, Italy.
Giordano
Assunta
A
Esposito
Luciana
L
Langella
Emma
E
Rossi
Mosè
M
Raia
Carlo A
CA
D019840:2-Propanol / Q000737:chemistry; D000426:Alcohol Dehydrogenase / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D003903:Deuterium; D008958:Models, Molecular; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D013237:Stereoisomerism; D016963:Thermus thermophilus / Q000201:enzymology*
D001426:Bacterial Proteins; D009243:NAD; D003903:Deuterium; D000426:Alcohol Dehydrogenase; D019840:2-Propanol