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Alcohol dehydrogenase in non-aqueous media using high-pressure technologies : reaction set-up and deactivation determination
Thorey, Paul ; Knez, Željko ; Leitgeb, Maja
BACKGROUND: The demand for enantiomerically pure molecules is growing continuously and biocatalysis is a powerful technique for their production. Inthis work, the catalyst was an enzyme combined with ...its coenzyme, NADP. Highpressure technology, a second clean technology, was applied as well. Densegases are promising solvents for biocatalysis. They have been investigated extensively as reaction media for lipase-catalysed reactions, butseldom for reactions, catalysed with alcohol dehydrogenases, as in this work. RESULT: The production of optically pure R-1-phenylethanol from acetophenone was investigated. The NADP-dependent alcohol dehydrogenase from Lactobacillus brevis (LBADH) was used as a catalyst. The hydrogenation was performed with isopropanol as a co-substrate in different conditions: dense propane with LBADH and NADP co-immobilized on glass beads and in the biphasic system water/dense propane. The obtained R-1-phenylethanol was enantiopure. The conversions were up to 90%. Deactivation of LBADH was also measured in these media. CONCLUSION: Protocols were successfully developed for the testing of alcohol dehydrogenase activity in dense gases. Enantioselectivity of LBADH is excellent in those media but it deactivated quickly. An LBADH-catalysed reaction was performed in a dense gas for the first time.
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