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Literature summary extracted from

  • Machielsen, R.; Looger, L.; Raedts, J.; Dijkhuizen, S.; Hummel, W.; Henneman, H.; Daussmann, T.; van der Oost, J.
    Cofactor engineering of Lactobacillus brevis alcohol dehydrogenase by computational design (2009), Eng. Life Sci., 9, 38-44.
No PubMed abstract available

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.B4 expression in Escherichia coli Levilactobacillus brevis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.B4 A90S activity with NADPH as cofactor decreased to about 50% of wild-type, activity with NADH strongly decreased Levilactobacillus brevis
1.1.1.B4 G37D/R38P activity with NADH is decreased relative to Arg38Pro alone, but is higher than that of the wild-type enzyme Levilactobacillus brevis
1.1.1.B4 M140I no activity with NADPH as cofactor Levilactobacillus brevis
1.1.1.B4 R38P no activity with NADPH as cofactor, fourfold increase in activity with NADH Levilactobacillus brevis
1.1.1.B4 V112D no activity with NADPH as cofactor, strongly decreased activity with NADH Levilactobacillus brevis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.B4 0.017
-
 NADPH wild-type, pH 6.5, 30°C Levilactobacillus brevis
1.1.1.B4 0.028
-
 NADPH mutant R38P, pH 5.5, 30°C Levilactobacillus brevis
1.1.1.B4 0.12
-
 NADH mutant R38P, pH 5.5, 30°C Levilactobacillus brevis
1.1.1.B4 0.13
-
 NADH wild-type, pH 6.5, 30°C Levilactobacillus brevis

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.B4 Levilactobacillus brevis
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.B4 acetophenone + NADH + H+
-
 Levilactobacillus brevis phenylethanol + NAD+
-
 ?
1.1.1.B4 acetophenone + NADPH + H+
-
 Levilactobacillus brevis phenylethanol + NADP+
-
 ?

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.B4 30
-
 wild-type, half-life 160 h, mutant R38P, half-life 220 h Levilactobacillus brevis
1.1.1.B4 42
-
 wild-type, half-life 1.25 h, mutant R38P, half-life 3 h Levilactobacillus brevis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.B4 5.5
-
 wild-type, cofactor NADH. Mutant R38P, both cofactors NADH and NADPH Levilactobacillus brevis
1.1.1.B4 6 6.5 wild-type, cofactor NADPH Levilactobacillus brevis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.B4 NADPH clear preference for NADPH as cofactor. The phosphate group of NADPH interacts with both the side chains and main-chain amides of Arg38 and Thr15, stabilizing this cofactor significantly better than NADH Levilactobacillus brevis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.B4 75
-
 NADH wild-type, pH 6.5, 30°C Levilactobacillus brevis
1.1.1.B4 300
-
 NADH mutant R38P, pH 5.5, 30°C Levilactobacillus brevis
1.1.1.B4 5100
-
 NADPH mutant R38P, pH 5.5, 30°C Levilactobacillus brevis
1.1.1.B4 9300
-
 NADPH wild-type, pH 6.5, 30°C Levilactobacillus brevis