Effects of temperature and pressure on enantiospecificity of novel secondary alcohol dehydrogenase mutants from Thermoanaerobacter ethanolicus
Patel, Jay Mayank
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Alcohol dehydrogenases (ADHs) are enzymes that catalyze the reversible reduction of carbonyl compounds to their corresponding alcohols. In chapter 2, we study the effect of hydrostatic pressure on stereospecificity of secondary ADH (SADH) from Thermoanaerobacter ethanolicus catalyzed oxidation of alcohols. Under high pressure conditions of 137.5 MPa and at 298K, the enantiomeric ratio (E) can be enhanced to 13.5 compared to 3.9 at room temperature and pressure for (S)-2-hexanol over (R)-2-hexanol. In chapter 3, site saturation mutagenesis approach was adopted in creating a comprehensive SADH mutant library at W110; and we used phenylacetone as a model substrate to study the effectiveness of our library. We are pleased to note that five of our mutants gave reductions at >99.9% e.e. and two of the mutants showed an E of over 100 for (S)-1-phenyl-2-propanol.