1.1.1.B20: meso-2,3-butandiol dehydrogenase

This is an abbreviated version, for detailed information about meso-2,3-butandiol dehydrogenase, go to the full flat file.

Reaction

(2R,3S)-butane-2,3-diol
+
NAD+
=
acetoin
+
NADH
+
H+

Synonyms

2,3-BD dehydrogenase, 2,3-butanediol dehydrogenae, ADH-9, ARA1, BDH, budC, butACg, mbdh, meso-2,3-BD dehydrogenase, meso-2,3-BDH, meso-2,3-butanediol dehydrogenase, meso-acetoin reductase, meso-BD, meso-BDH, NAD(H)-dependent meso-2,3-BDH, NAD(H)-dependent meso-2,3-butanediol 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.B20 meso-2,3-butandiol dehydrogenase

Engineering

Engineering on EC 1.1.1.B20 - meso-2,3-butandiol dehydrogenase

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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D194G
site-directed mutagenesis, the mutant binds the substrate but is catalytically almost inactive. The mutant is inactive with (2S,3S)-butanediol, meso-butanediol and (2R,3R)-butanediol. D194G enzyme mutant shows a similar secondary structure compared to Enterobacter aerogenes BDH. While the mutant is highly susceptible to protease digestion compared to the wild-type enzyme. Homology modeling of the mutant enzyme, with meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae, PDB ID 1GEG, as a template, reveals that Gly194 seems to lose the hydrogen bond interactions with the surrounding residues (Gly206, Gly207 and Thr209), resulting in a putative conformational changes of mutant D194G which might be responsible for the loss of activity
moe
-
construction and engineering of Corynebacterium glutamicum strain DELTAaceEDELTApqoDELTAldhA(pEKEx2-als,aldB,butACg). Chromosomal inactivation of the putative BDH gene butACg (cg2958) in strain DELTAaceEDELTApqoDELTAldhA. BDH activity is nearly abolished upon inactivation of butACg indicating that Corynebacterium glutamicum expresses a single BDH under the experimental conditions examined. BDH activity increases 3fold in strain DELTAaceEDELTApqoDELTAldhA(pEKEx2-als,aldB,butACg) compared to the respective control. The inactivation of butACg gene decreases the BDH activity 75fold for the DELTAaceEDELTApqoDELTAldhADELTAbutACg(pEKEx2) strain compared to strain DELTAaceEDELTApqoDELTAldhA(pEKEx2). The major form of 2,3-butanediol is meso-2,3-butandediol, and the ratio meso-2,3-BD/optically active 2,3-BD is 95:5, the main side products are glycerol, ethanol, and acetoin
N146F
-
11% of wild-type specific activity
Q140I
-
trace activity below 0.1 U/mg
Q140I/N146F
-
poor activity
Q140I/N146F/W190H
-
trace activity below 0.1 U/mg with substrate meso-butanediol, 2.9 U/mg with substrate (2S,3S)-2,3-butanediol
Q140I
-
mutation mimicking the corresponding residue in (S,S)-butanediol dehydrogenase. No activity with substrates meso-butanediol or (S,S)-butanediol
Q140I/N146F
-
mutation mimicking the corresponding residues in (S,S)-butanediol dehydrogenase. Poor activity with substrates meso-butanediol or (S,S)-butanediol
additional information