Deracemization of Secondary Alcohols by using a Single Alcohol Dehydrogenase

ABSTRACT The gene encoding a novel alcohol dehydrogenase that belongs to the aldo-keto reductase superfamily has been identified in the hyperthermophilic archaeon Pyrococcus furiosus. The gene, referred to as adhD, was functionally expressed in Escherichia coli and subsequently purified to homogeneity. The enzyme has a monomeric conformation with a molecular mass of 32 kDa. The catalytic activity of the enzyme increases up to 100°C, and a half-life value of 130 min at this temperature indicates its high thermostability. AdhD exhibits a broad substrate specificity with, in general, a preference for the reduction of ketones (pH optimum, 6.1) and the oxidation of secondary alcohols (pH optimum, 8.8). Maximal specific activities were detected with 2,3-butanediol (108.3 U/mg) and diacetyl-acetoin (22.5 U/mg) in the oxidative and reductive reactions, respectively. Gas chromatrography analysis indicated that AdhD produced mainly (S)-2-pentanol (enantiomeric excess, 89%) when 2-pentanone was used as substrate. The physiological role of AdhD is discussed.

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Chemoenzymatic Deracemization of Secondary Alcohols by using a TEMPO-Iodine-Alcohol Dehydrogenase System

ChemCatChem ◽

10.1002/cctc.201500816 ◽

2015 ◽

Vol 7 (24) ◽

pp. 4016-4020 ◽

Cited By ~ 14

Author(s):

Daniel Méndez-Sánchez ◽

Juan Mangas-Sánchez ◽

Iván Lavandera ◽

Vicente Gotor ◽

Vicente Gotor-Fernández

Keyword(s):

Alcohol Dehydrogenase ◽

Secondary Alcohols ◽

Dehydrogenase System

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Thermostable Alcohol Dehydrogenase from Thermococcus kodakarensis KOD1 for Enantioselective Bioconversion of Aromatic Secondary Alcohols

Applied and Environmental Microbiology ◽

10.1128/aem.03873-12 ◽

2013 ◽

Vol 79 (7) ◽

pp. 2209-2217 ◽

Cited By ~ 17

Author(s):

Xi Wu ◽

Chong Zhang ◽

Izumi Orita ◽

Tadayuki Imanaka ◽

Toshiaki Fukui ◽

...

Keyword(s):

Alcohol Dehydrogenase ◽

Enantiomeric Excess ◽

Chiral Alcohols ◽

Secondary Alcohols ◽

Optimal Temperature ◽

Content Type ◽

Hyperthermophilic Archaeon ◽

Thermococcus Kodakarensis ◽

Highly Active ◽

Thermococcus Kodakarensis Kod1

ABSTRACTA novel thermostable alcohol dehydrogenase (ADH) showing activity toward aromatic secondary alcohols was identified from the hyperthermophilic archaeonThermococcus kodakarensisKOD1 (TkADH). The gene,tk0845, which encodes an aldo-keto reductase, was heterologously expressed inEscherichia coli. The enzyme was found to be a monomer with a molecular mass of 31 kDa. It was highly thermostable with an optimal temperature of 90°C and a half-life of 4.5 h at 95°C. The apparentKmvalues for the cofactors NAD(P)+and NADPH were similar within a range of 66 to 127 μM.TkADH preferred secondary alcohols and accepted various ketones and aldehydes as substrates. Interestingly, the enzyme could oxidize 1-phenylethanol and its derivatives having substituents at themetaandparapositions with high enantioselectivity, yielding the corresponding (R)-alcohols with optical purities of greater than 99.8% enantiomeric excess (ee).TkADH could also reduce 2,2,2-trifluoroacetophenone to (R)-2,2,2-trifluoro-1-phenylethanol with high enantioselectivity (>99.6% ee). Furthermore, the enzyme showed high resistance to organic solvents and was particularly highly active in the presence of H2O–20% 2-propanol and H2O–50%n-hexane orn-octane. This ADH is expected to be a useful tool for the production of aromatic chiral alcohols.

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Enantioselective oxidation of secondary alcohols at a quinohaemoprotein alcohol dehydrogenase electrode

Applied Biochemistry and Biotechnology ◽

10.1007/bf02787770 ◽

1998 ◽

Vol 75 (2-3) ◽

pp. 151-162 ◽

Cited By ~ 5

Author(s):

Wim A. C. Somers ◽

Edwin C. A. Stigter ◽

Wim van Hartingsveldt ◽

Jan Pieter van der Lugt

Keyword(s):

Alcohol Dehydrogenase ◽

Secondary Alcohols ◽

Enantioselective Oxidation

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