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Literature summary for 1.8.1.4 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
Search for more literature for 1.8.1.4

Activating Compound

Activating Compound Comment Organism Structure
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)

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

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0061
-
 NADH in 50 mM potassium phosphate buffer (pH 6.0), at 25°C Microbacterium luteolum
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]

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

Organism

Organism UniProt Comment Textmining
Microbacterium luteolum B6F1A8
-
-
Microbacterium luteolum JCM 9174 B6F1A8
-
-

Purification (Commentary)

Purification (Comment) Organism
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]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
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
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)

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

Subunits

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

Synonyms

Synonyms Comment Organism
dihydrolipoamide dehydrogenase
-
 Microbacterium luteolum
LPD
-
 Microbacterium luteolum

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
 Microbacterium luteolum

pH Range

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

Cofactor

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

General Information

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