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Literature summary extracted from

  • Pennacchio, A.; Sannino, V.; Sorrentino, G.; Rossi, M.; Raia, C.A.; Esposito, L.
    Biochemical and structural characterization of recombinant short-chain NAD(H)-dependent dehydrogenase/reductase from Sulfolobus acidocaldarius highly enantioselective on diaryl diketone benzil (2013), Appl. Microbiol. Biotechnol., 97, 3949-3964.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.1 heterologously overexpressed in Escherichia coli Sulfolobus acidocaldarius

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.1.1.1 the crystal structure of the binary complex SaADH2–NADH, determined at 1.75 A resolution, reveals details of the active site providing hints on the structural basis of the enzyme enantioselectivity Sulfolobus acidocaldarius

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.1 1-butyl-3-methylimidazolium tetrafluoroborate 5%, 50% inhibition, presumably due to a competition of the BF4- ion with the coenzyme phosphate moiety for the anion-binding site of the enzyme Sulfolobus acidocaldarius
1.1.1.1 Fe2+ 1 mM, 7% inhibition Sulfolobus acidocaldarius

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.1 0.04
-
 NADH 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.11
-
 2,2-dichloroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.18
-
 NAD+ 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 0.43
-
 benzil 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.81
-
 isoborneol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 4.2
-
 ethyl benzoylformate 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 5
-
 1-phenyl-1,2-propanedione 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 5.1
-
 cycloheptanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 6.3
-
 2,2,2-trifluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 8.5
-
 tetralin-1-ol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 8.7
-
 1-Indanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 11.9
-
 2',3',4',5',6'-pentafluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 16.6
-
 ethyl 3-methyl-2-oxobutyrate 65°C, pH 5.0 Sulfolobus acidocaldarius

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.1.1.1 Li+ 1 mM, 1.15fold activation Sulfolobus acidocaldarius
1.1.1.1 Mn2+ 1 mM, 1.1 fold activation Sulfolobus acidocaldarius
1.1.1.1 Na+ 1 mM, 1.13fold activation Sulfolobus acidocaldarius

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.1 27024
-
 4 * 27024, ESI-MS analysis Sulfolobus acidocaldarius
1.1.1.1 29000
-
 4 * 29000, SDS-PAGE Sulfolobus acidocaldarius
1.1.1.1 109000
-
 sucrose density gradient centrifugation Sulfolobus acidocaldarius

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.1 Sulfolobus acidocaldarius Q4J9F2
-
-
1.1.1.1 Sulfolobus acidocaldarius DSM 639 Q4J9F2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.1
-
 Sulfolobus acidocaldarius

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.1 1-decalone + NADH + H+ 85% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius decahydronaphthalen-1-ol + NAD+
-
 ?
1.1.1.1 1-indanol + NAD+ 26% of the activity compared to isoborneol Sulfolobus acidocaldarius 1-indanone + NADH + H+
-
 ?
1.1.1.1 1-indanol + NAD+ 26% of the activity compared to isoborneol Sulfolobus acidocaldarius DSM 639 1-indanone + NADH + H+
-
 ?
1.1.1.1 1-phenyl-1,2-propanedione + NADH + H+ 1-phenyl-1,2-propanedione and ethyl 3-methyl-2-oxobutyrate are the best substrate in the reduction reaction Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 1-phenyl-1,2-propanedione + NADH + H+ 1-phenyl-1,2-propanedione and ethyl 3-methyl-2-oxobutyrate are the best substrate in the reduction reaction Sulfolobus acidocaldarius DSM 639 ?
-
 ?
1.1.1.1 2',3',4',5',6'-pentafluoroacetophenone + NADH + H+ 45% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 1-(2,3,4,5,6-pentafluorophenyl)ethanol + NAD+
-
 ?
1.1.1.1 2,2'-dichlorobenzil + NADH + H+
-
 Sulfolobus acidocaldarius 1,2-bis(2-chlorophenyl)-2-hydroxyethanone + NAD+
-
 ?
1.1.1.1 2,2,2-trifluoroacetophenone + NADH + H+ 35% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 2,2,2-trifluoro-1-phenylethanol + NAD+
-
 ?
1.1.1.1 2,2-dichloroacetophenone + NADH + H+ 30% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 2,2-dichloro-1-phenylethanol + NAD+
-
 ?
1.1.1.1 2-methylcyclohexanone + NADH + H+ 13% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 2-methylcyclohexanol + NAD+
-
 ?
1.1.1.1 3-methylcyclohexanol + NAD+ 10% of the activity compared to isoborneol Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 3-methylcyclohexanone + NADH + H+ 33% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 3-methylcyclohexanol + NAD+
-
 ?
1.1.1.1 4-methylcyclohexanone + NADH + H+ 60% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius 4-methylcyclohexanol + NAD+
-
 ?
1.1.1.1 acetophenone + NADH + H+
-
 Sulfolobus acidocaldarius 1-phenylethanol + NAD+
-
 ?
1.1.1.1 acetophenone + NADH + H+
-
 Sulfolobus acidocaldarius DSM 639 1-phenylethanol + NAD+
-
 ?
1.1.1.1 benzil + NADH + H+ 62% of the activity compared to 1-phenyl-1,2-propanedione. The enzyme catalyses the asymmetric reduction of benzil to (R)-benzoin with both excellent conversion (98%) and optical purity (98%) by way of an efficient in situ NADH-recycling system involving a second thermophilic ADH Sulfolobus acidocaldarius (R)-benzoin + NAD+
-
 ?
1.1.1.1 cycloheptanol + NAD+
-
 Sulfolobus acidocaldarius cycloheptanone + NADH + H+
-
 ?
1.1.1.1 cyclohexanone + NADH + H+
-
 Sulfolobus acidocaldarius cyclohexanol + NAD+
-
 ?
1.1.1.1 cyclohexanone + NADH + H+
-
 Sulfolobus acidocaldarius DSM 639 cyclohexanol + NAD+
-
 ?
1.1.1.1 cyclopentanol + NAD+ 55% of the activity compared to isoborneol Sulfolobus acidocaldarius cyclopentanone + NADH + H+
-
 ?
1.1.1.1 ethyl 3-methyl-2-oxobutyrate + NADH + H+ 1-phenyl-1,2-propanedione and ethyl 3-methyl-2-oxobutyrate are the best substrate in the reduction reaction Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 ethyl benzoylformate + NADH + H+ 23% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 ethyl pyruvate + NADH + H+ 16% of the activity compared to 1-phenyl-1,2-propanedione Sulfolobus acidocaldarius ethyl 2-hydroxypropionate + NAD+
-
 ?
1.1.1.1 isoborneol + NAD+ isoborneol is the best substrate in the oxidation reaction Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 additional information the enzyme displays a preference for the reduction of alicyclic, bicyclic and aromatic ketones and alpha-ketoesters, but is poorly active on aliphatic, cyclic and aromatic alcohols, showing no activity on aldehydes Sulfolobus acidocaldarius ?
-
 ?
1.1.1.1 additional information the enzyme displays a preference for the reduction of alicyclic, bicyclic and aromatic ketones and alpha-ketoesters, but is poorly active on aliphatic, cyclic and aromatic alcohols, showing no activity on aldehydes Sulfolobus acidocaldarius DSM 639 ?
-
 ?
1.1.1.1 tetralin-1-ol + NAD+ 51% of the activity compared to isoborneol Sulfolobus acidocaldarius 3,4-dihydronaphthalen-1(2H)-one + NADH + H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.1 tetramer 4 * 29000, SDS-PAGE Sulfolobus acidocaldarius
1.1.1.1 tetramer 4 * 27024, ESI-MS analysis Sulfolobus acidocaldarius

Synonyms

EC Number Synonyms Comment Organism
1.1.1.1 SaADH2
-
 Sulfolobus acidocaldarius

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.1 65
-
 assay at Sulfolobus acidocaldarius
1.1.1.1 78
-
 the reaction rate increases up to 78°C and then decreases rapidly due to thermal inactivation Sulfolobus acidocaldarius

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.1 65 85 65°C: about 60% of maximal activity, 85°C: about 70% of maximal activity Sulfolobus acidocaldarius

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.1 75
-
 30 min, stable up to Sulfolobus acidocaldarius
1.1.1.1 88
-
 30-min half-inactivation temperature Sulfolobus acidocaldarius

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.1.1.1 0.65
-
 2,2-dichloroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 1.6
-
 benzil 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 1.7
-
 2,2,2-trifluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 1.9
-
 ethyl benzoylformate 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 3.2
-
 2',3',4',5',6'-pentafluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 5.3
-
 1-phenyl-1,2-propanedione 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 6.2
-
 1-Indanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 7
-
 cycloheptanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 9.6
-
 tetralin-1-ol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 16.6
-
 isoborneol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 19.3
-
 NAD+ 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 26
-
 ethyl 3-methyl-2-oxobutyrate 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 26.2
-
 NADH 65°C, pH 5.0 Sulfolobus acidocaldarius

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.1 5
-
 ketone reduction reaction Sulfolobus acidocaldarius
1.1.1.1 10
-
 alcohol oxidation reaction Sulfolobus acidocaldarius

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.1 NAD+ strict requirement for NAD(H) as the coenzyme, no activity with NADP+. The specificity constant value is 6fold higher for NADH than NAD+ Sulfolobus acidocaldarius
1.1.1.1 NADH strict requirement for NAD(H) as the coenzyme, no activity with NADPH. The specificity constant value is 6fold higher for NADH than NAD+ Sulfolobus acidocaldarius

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.1.1.1 0.27
-
 2,2,2-trifluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.27
-
 2',3',4',5',6'-pentafluoroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.45
-
 ethyl benzoylformate 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 0.71
-
 1-Indanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 1.06
-
 1-phenyl-1,2-propanedione 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 1.13
-
 tetralin-1-ol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 1.37
-
 cycloheptanol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 1.57
-
 ethyl 3-methyl-2-oxobutyrate 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 3.72
-
 benzil 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 5.91
-
 2,2-dichloroacetophenone 65°C, pH 5.0 Sulfolobus acidocaldarius
1.1.1.1 20.5
-
 isoborneol 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 107
-
 NAD+ 65°C, pH 10.5 Sulfolobus acidocaldarius
1.1.1.1 655
-
 NADH 65°C, pH 5.0 Sulfolobus acidocaldarius