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Literature summary extracted from

  • Kurokawa, J.; Asano, M.; Nomoto, S.; Makino, Y.; Itoh, N.
    Gene cloning and characterization of dihydrolipoamide dehydrogenase from Microbacterium luteolum: A useful enzymatic regeneration system of NAD+ from NADH (2010), J. Biosci. Bioeng., 109, 218-223.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.8.1.4 NAD+ 0.2 mM NAD+ demonstrates a strong activating effect on LPD, and the activity is 5.2 times higher than that without NAD+ Microbacterium luteolum

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.8.1.4 expressed in Escherichia coli JM109 cells Microbacterium luteolum

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8.1.4 0.0061
-
 NADH in 50 mM potassium phosphate buffer (pH 6.0), at 25°C Microbacterium luteolum
1.8.1.4 3.3
-
 Lipoamide in the presence of 0.2 mM NAD+, in 50 mM potassium phosphate buffer (pH 6.0), at 25°C Microbacterium luteolum

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.8.1.4 49912
-
 2 * 49912, calculated from amino acid sequence Microbacterium luteolum
1.8.1.4 50000
-
 2 * 50000, SDS-PAGE Microbacterium luteolum
1.8.1.4 102000
-
 native enzyme, gel filtration Microbacterium luteolum

Organism

EC Number Organism UniProt Comment Textmining
1.8.1.4 Microbacterium luteolum B6F1A8
-
-
1.8.1.4 Microbacterium luteolum JCM 9174 B6F1A8
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.8.1.4 DEAE-Toyopearl 650 M column chromatography, butyl-Toyopearl 650 M column chromatography, and TSK-gel G3000SW gel filtration Microbacterium luteolum

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.8.1.4 2.5
-
 crude extract, in the presence of 0.2 mM NAD+, in 50 mM potassium phosphate buffer (pH 6.0), at 25°C Microbacterium luteolum
1.8.1.4 59.2
-
 after 23.9fold purification, in the presence of 0.2 mM NAD+, in 50 mM potassium phosphate buffer (pH 6.0), at 25°C Microbacterium luteolum

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.8.1.4 1-methoxyphenazinium methosulfate + NADH 248.1% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 1-methoxyphenazinium methosulfate + NADH 248.1% activity compared to lipoamide Microbacterium luteolum JCM 9174 ? + NAD+
-
 ?
1.8.1.4 2,6-dimethoxy-1,4-benzoquinone + NADH 8.0% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 ferrocene + NADH 3.6% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 ferrocenecarboxylic acid + NADH 4.1% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 iodonitrotetrazolium chloride + NADH 19.3% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 lipoamide + NADH 100% activity Microbacterium luteolum dihydrolipoamide + NAD+
-
 ?
1.8.1.4 lipoamide + NADH 100% activity Microbacterium luteolum JCM 9174 dihydrolipoamide + NAD+
-
 ?
1.8.1.4 menadione + NADH 2.9% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 menadione + NADH 2.9% activity compared to lipoamide Microbacterium luteolum JCM 9174 ? + NAD+
-
 ?
1.8.1.4 additional information no activity with NADPH, potassium ferricyanide and methylene blue Microbacterium luteolum ?
-
 ?
1.8.1.4 additional information no activity with NADPH, potassium ferricyanide and methylene blue Microbacterium luteolum JCM 9174 ?
-
 ?
1.8.1.4 nitrotetrazolium blue + NADH 2.9% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 oxidized 2,6-dichlorophenolindophenol + NADH 233.4% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 oxidized 2,6-dichlorophenolindophenol + NADH 233.4% activity compared to lipoamide Microbacterium luteolum JCM 9174 ? + NAD+
-
 ?
1.8.1.4 resazurin + NADH 155.8% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 resorufin + NADH 19.1% activity compared to lipoamide Microbacterium luteolum ? + NAD+
-
 ?
1.8.1.4 sulfonated tetrazolium WST-1 + NADH 39.2% activity compared to lipoamide Microbacterium luteolum formazan derivative + NAD+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.8.1.4 homodimer 2 * 50000, SDS-PAGE Microbacterium luteolum
1.8.1.4 homodimer 2 * 49912, calculated from amino acid sequence Microbacterium luteolum

Synonyms

EC Number Synonyms Comment Organism
1.8.1.4 dihydrolipoamide dehydrogenase
-
 Microbacterium luteolum
1.8.1.4 LPD
-
 Microbacterium luteolum

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.8.1.4 55 70 the remaining activity after thermal treatment at pH 7.0 for 15 min is 90% at 55°C, 80% at 60°C, 70% at 65 °C, and 45% at 70°C Microbacterium luteolum

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.8.1.4 6
-
-
 Microbacterium luteolum

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.8.1.4 6 7 high activity in a pH range of 6.0 to 7.0 Microbacterium luteolum

Cofactor

EC Number Cofactor Comment Organism Structure
1.8.1.4 FAD 1.34 mol FAD per 1 mol of subunit of the enzyme Microbacterium luteolum
1.8.1.4 NADH high affinity for NADH, no activity with NADPH Microbacterium luteolum

General Information

EC Number General Information Comment Organism
1.8.1.4 physiological function LPD is a useful biocatalyst for regenerating NAD+ Microbacterium luteolum