1.1.1.B4: (R)-specific secondary alcohol dehydrogenase (NADH)

This is an abbreviated version, for detailed information about (R)-specific secondary alcohol dehydrogenase (NADH), go to the full flat file.

Reaction

(R)-R-CHOH-R'
+
NAD+
=
R-CO-R'
+
NADH
+
H+

Synonyms

(R)-epinephrine dehydrogenase, ADH, ADH1, ADH2, ADHTt, carbonyl reductase, carbonyl reductase (NADH, specific for (S)-configuration of alcohol), CHY1186, CpCR, mLDH, More, SADH, SDR, secondary alcohol dehydrogenase, short-chain NAD(H)-dependent alcohol dehydrogenase

ECTree

     1 Oxidoreductases
         1.1 Acting on the CH-OH group of donors
             1.1.1 With NAD+ or NADP+ as acceptor
                1.1.1.B4 (R)-specific secondary alcohol dehydrogenase (NADH)

Engineering

Engineering on EC 1.1.1.B4 - (R)-specific secondary alcohol dehydrogenase (NADH)

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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A90S
-
activity with NADPH as cofactor decreased to about 50% of wild-type, activity with NADH strongly decreased
G37D/R38P
-
activity with NADH is decreased relative to Arg38Pro alone, but is higher than that of the wild-type enzyme
M140I
-
no activity with NADPH as cofactor
R38P
-
no activity with NADPH as cofactor, fourfold increase in activity with NADH
V112D
-
no activity with NADPH as cofactor, strongly decreased activity with NADH
S154Y
mutant exhibits nearly 13fold, 5.4fold, and 2.3fold increase in kcat/Km value, kcat value, and specific activity toward 3,5-bis(trifluoromethyl)acetophenone
S154Y/L194I
180% of wild-type activity
S154Y
-
mutant exhibits nearly 13fold, 5.4fold, and 2.3fold increase in kcat/Km value, kcat value, and specific activity toward 3,5-bis(trifluoromethyl)acetophenone
-
S154Y/L194I
-
180% of wild-type activity
-
W95L/N249Y
-
the mutant exhibits higher activity but decreased affinity toward aliphatic alcohols, aldehydes as well as NAD+ and NADH compared to the wild type enzyme, optimum pH is at about pH 8.6
C295A
-
the mutant shows an increased size of the alkyl group which can bind in the substrate small pocket by one carbon atom compared to the wild-type enzyme
I86A/C295A
-
the mutant enzyme shows broadened substrate specificity for aryl ketones and broadened substrate specificity for meta-substituted, but not para-substituted, acetophenones compared to the wild-type enzyme. The increase of the substrate specificity of I86A/C295A SADH is accompanied by a decrease in the kcat/Km values of acetophenones, possibly due to the substrates fitting loosely inside the more open active site