Biocatalysis, Department of Biotechnology, Delft University of Technology, Julianalaan 136, Delft, 2628 BL, The Netherlands.
AMB Express. 2014 Mar 15;4:30. doi: 10.1186/s13568-014-0030-2. eCollection 2014.
The Michael hydratase - alcohol dehydrogenase (MhyADH) from Alicycliphilus denitrificans was previously identified as a bi-functional enzyme performing a hydration of α,β-unsaturated ketones and subsequent oxidation of the formed alcohols. The investigations of the bi-functionality were based on a spectrophotometric assay and an activity staining in a native gel of the dehydrogenase. New insights in the recently discovered organocatalytic Michael addition of water led to the conclusion that the previously performed experiments to identify MhyADH as a bi-functional enzyme and their results need to be reconsidered and the reliability of the methodology used needs to be critically evaluated.
先前从脱硫球形菌（Alicycliphilus denitrificans）中鉴定出 Michael 水解酶-醇脱氢酶（MhyADH）为一种双功能酶，可使α,β-不饱和酮水合，并随后氧化形成的醇。双功能的研究是基于分光光度法测定和脱氢酶的天然胶中活性染色。在最近发现的有机催化的水合 Michael 加成反应方面的新见解得出的结论是，先前进行的将 MhyADH 鉴定为双功能酶的实验及其结果需要重新考虑，并且需要严格评估所使用方法的可靠性。
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