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Literature summary extracted from

  • Schlieben, N.H.; Niefind, K.; Mueller, J.; Riebel, B.; Hummel, W.; Schomburg, D.
    Atomic resolution of R-specific alcohol dehydrogenase from Lactobacillus brevis provide the structual bases of its substrate and cosubstrate specificity (2005), J. Mol. Biol., 349, 801-813.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.2 cloned into the pkk177-3H vector, transformation in Escherichia coli Levilactobacillus brevis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.2 16°C, wild-type RADH: 15 mg/ml stock solution (21% polyethyleneglycol monomethyl ether 550, 0.1 M Tris-HCl, 50 nM MgCl2, 40 mM acetophenone, 10 mM NADP+), growth for 3 weeks, max. size 0.25 mm x 0.25 mm x 1.0 mm, RADH-G37D: 21 mg/ml stock solution (methyl-2,4-pentanediol, 20% polyethyleneglycol 400 or polyethyleneglycol monomethyl ether 550, 0.1 M Hepes, 50 mM MgCl2 59 mM 1-phenylethanol, 25 mM NADH), growth 1 week, max. size 0.4 mm x 0.5 mm x 1.7 mm Levilactobacillus brevis

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.2 G37D regular PCR with site-specific mutation for both DNA strands, the gene is divided into two separate amplification products, preference for NAD+ rather than NADP+, depends on an a interaction between the adenosine ribose moiety of NAD+ and the inserted aspartate side chain Levilactobacillus brevis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.2 0.015
-
 NADP+ reduction, recombinant wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 0.04
-
 NADPH oxidation, recombinante wild-typ, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 0.75
-
 NAD+ reduction, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 1.6
-
 NADH oxidation, recombinante mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 2.8
-
 acetophenone cofactor NADPH, reduction, recombinante wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 2.9
-
 1-phenylethanol cofactor NADP+, oxidation, recombinate wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 9.1
-
 NADP+ reduction, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 10.8
-
 1-phenylethanol NAD+, oxidation, recombinante mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 10.9
-
 acetophenone cofactor NADH, reduction, recombinante mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 55.5
-
 1-phenylethanol cofactor NADP+, oxidation, recombinate mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.2 Levilactobacillus brevis
-
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.2 acetophenone + NADH + H+ recombinant mutant G37D Levilactobacillus brevis 1-phenylethanol + NAD+ recombinate mutant G37D r
1.1.1.2 acetophenone + NADPH + H+ recombinant wild-type Levilactobacillus brevis 1-phenylethanol + NADP+ recombinant wild-type r
1.1.1.2 phenylethanol + NADP+
-
 Levilactobacillus brevis phenylacetaldehyde + NADPH + H+
-
 r

Synonyms

EC Number Synonyms Comment Organism
1.1.1.2 RADH
-
 Levilactobacillus brevis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.2 30
-
-
 Levilactobacillus brevis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.2 4.4
-
 NADP+ reduction, recombinant wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 5.4
-
 Phenylethanol oxidation, cosubstrate NADP+, recombinant wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 5.9
-
 NADP+ reduction, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 8.2
-
 Phenylethanol oxidation, cosubstrate NADP+, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 9.5
-
 Phenylethanol oxidation, cosubstrate NAD+, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 9.9
-
 NAD+ reduction, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 35.7
-
 acetophenone cofactor NADH, reduction, recombinant mutant G37Dm 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 36.6
-
 NADH oxidation, recombinant mutant G37D, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 38.1
-
 NADPH oxidation, recombinant wild-type, 30°C, pH 7.0 Levilactobacillus brevis
1.1.1.2 44.5
-
 acetophenone cofactor NADPH, reduction, recombinant wild-type, 30°C, pH 7.0 Levilactobacillus brevis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.2 7
-
-
 Levilactobacillus brevis

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.2 NAD+
-
 Levilactobacillus brevis
1.1.1.2 NADH
-
 Levilactobacillus brevis
1.1.1.2 NADP+
-
 Levilactobacillus brevis
1.1.1.2 NADPH
-
 Levilactobacillus brevis