Skip to Content
Merck

Michael hydratase alcohol dehydrogenase or just alcohol dehydrogenase?

AMB Express (2014-06-21)
Verena Resch, Jianfeng Jin, Bi-Shuang Chen, Ulf Hanefeld
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
2-Cyclohexen-1-one, ≥95%
Sigma-Aldrich
Potassium nitrate, 99.99% trace metals basis
Sigma-Aldrich
Potassium nitrate, 99.999% trace metals basis
Sigma-Aldrich
2,6-Dichlorophenol, 99%
Sigma-Aldrich
Potassium nitrate, BioReagent, suitable for cell culture, suitable for plant cell culture
Sigma-Aldrich
Nitrotetrazolium Blue chloride, ≥90.0% (HPLC)
Sigma-Aldrich
Nitrotetrazolium Blue chloride, powder, electrophoresis grade
Sigma-Aldrich
Potassium nitrate, BioXtra, ≥99.0%
Sigma-Aldrich
Potassium phosphate dibasic, anhydrous, for luminescence, for molecular biology, BioUltra, ≥99.0% (T)
Sigma-Aldrich
Potassium phosphate monobasic, 99.99% trace metals basis
Sigma-Aldrich
Calcium chloride, AnhydroBeads, −10 mesh, ≥99.9% trace metals basis
Sigma-Aldrich
Magnesium sulfate solution, BioUltra, for molecular biology
Sigma-Aldrich
Potassium phosphate dibasic, 99.95% trace metals basis
Sigma-Aldrich
Methylene Blue solution, 0.05 wt. % in H2O
Sigma-Aldrich
Calcium chloride, AnhydroBeads, −10 mesh, ≥99.99% trace metals basis
Sigma-Aldrich
Tetrazolium Blue Chloride, indicator for germination, suitable for microbiology, ≥90% (T)
Sigma-Aldrich
Calcium chloride, anhydrous, powder, 99.99% trace metals basis
Sigma-Aldrich
Magnesium sulfate, ≥99.99% trace metals basis
Sigma-Aldrich
Calcium chloride solution, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
Potassium phosphate monobasic, BioUltra, for molecular biology, anhydrous, ≥99.5% (T)
Sigma-Aldrich
Methylene Blue solution, for microscopy, concentrate according to Ehrlich, concentrated, aqueous solution
Sigma-Aldrich
Cyclohexanol, ReagentPlus®, 99%
Sigma-Aldrich
Xanthine, ≥99%
Sigma-Aldrich
Potassium phosphate dibasic, meets USP testing specifications
Sigma-Aldrich
Calcium chloride, anhydrous, BioReagent, suitable for insect cell culture, suitable for plant cell culture, ≥96.0%
Sigma-Aldrich
Calcium chloride
Sigma-Aldrich
Tetrazolium Blue Chloride, used in colorimetric determination of reducing compounds
Sigma-Aldrich
Magnesium sulfate solution, for molecular biology, 1.00 M±0.04 M
Sigma-Aldrich
Magnesium sulfate, BioReagent, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Potassium phosphate monobasic, for molecular biology, ≥98.0%