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Literature summary for 2.6.1.76 extracted from

  • Richter, A.A.; Mais, C.N.; Czech, L.; Geyer, K.; Hoeppner, A.; Smits, S.H.J.; Erb, T.J.; Bange, G.; Bremer, E.
    Biosynthesis of the stress-protectant and chemical chaperon ectoine biochemistry of the transaminase EctB (2019), Front. Microbiol., 10, 2811 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
gene ectB, genetic structure, recombinant overexpression of codon-optimized version of (Pl)ectB as N-terminally Strep II-tagged protein from modified plasmid pLC52 in Escherichia coli strain BL21 and TOP 10 cells Paenibacillus lautus

Protein Variants

Protein Variants Comment Organism
K274A site-directed mutagenesis, the mutant is unable to bind cofactor PLP and ins catalytically inactive Paenibacillus lautus
K274H site-directed mutagenesis, the mutant is unable to bind cofactor PLP and ins catalytically inactive. This amino acid substitution does not affect the quaternary assembly of the mutant protein Paenibacillus lautus
K274R site-directed mutagenesis, the mutant is unable to bind cofactor PLP and ins catalytically inactive Paenibacillus lautus

Inhibitors

Inhibitors Comment Organism Structure
KCl activates slightly at up to 350 mM, inhibits by 50% at 1.5 M Paenibacillus lautus
NaCl activates slightly at up to 350 mM, inhibits by 50% at 1.25 M Paenibacillus lautus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information Michaelis-Menten kinetics Paenibacillus lautus
0.43
-
L-2,4-diaminobutanoate pH 7.0, 45┬░C, recombinant enzyme Paenibacillus lautus
9.01
-
2-oxoglutarate pH 7.0, 45┬░C, recombinant enzyme Paenibacillus lautus

Metals/Ions

Metals/Ions Comment Organism Structure
KCl activates slightly at up to 350 mM, inhibits by 50% at 1.5 M Paenibacillus lautus
NaCl activates slightly at up to 350 mM, inhibits by 50% at 1.25 M Paenibacillus lautus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
200000
-
about, recombinant Strep II-tagged enzyme, gel filtration Paenibacillus lautus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-2,4-diaminobutanoate + 2-oxoglutarate Paenibacillus lautus
-
L-aspartate 4-semialdehyde + L-glutamate
-
?
L-2,4-diaminobutanoate + 2-oxoglutarate Paenibacillus lautus Y412MC10
-
L-aspartate 4-semialdehyde + L-glutamate
-
?

Organism

Organism UniProt Comment Textmining
Paenibacillus lautus D3EKC0 isolated from the effluent of a hot spring in the Yellowstone National Park (United States)
-
Paenibacillus lautus Y412MC10 D3EKC0 isolated from the effluent of a hot spring in the Yellowstone National Park (United States)
-

Purification (Commentary)

Purification (Comment) Organism
recombinant Strep II-tagged enzyme from Escherichia coli by streptavidin-affinity chromatography Paenibacillus lautus

Source Tissue

Source Tissue Comment Organism Textmining
cell culture growth at temperatures of up to 50┬░C Paenibacillus lautus
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Paenibacillus lautus L-aspartate 4-semialdehyde + L-glutamate
-
?
L-2,4-diaminobutanoate + 2-oxoglutarate preferred enzyme Paenibacillus lautus L-aspartate 4-semialdehyde + L-glutamate
-
?
L-2,4-diaminobutanoate + 2-oxoglutarate
-
Paenibacillus lautus Y412MC10 L-aspartate 4-semialdehyde + L-glutamate
-
?
L-2,4-diaminobutanoate + 2-oxoglutarate preferred enzyme Paenibacillus lautus Y412MC10 L-aspartate 4-semialdehyde + L-glutamate
-
?
additional information (Pl)EctB also catalyzes a GABA-TA-type reaction, EC 2.6.1.19, but with less activity Paenibacillus lautus ?
-
-
additional information (Pl)EctB also catalyzes a GABA-TA-type reaction, EC 2.6.1.19, but with less activity Paenibacillus lautus Y412MC10 ?
-
-

Subunits

Subunits Comment Organism
homotetramer 4 * 49100, recombinant Strep II-tagged enzyme, SDS-PAGE Paenibacillus lautus

Synonyms

Synonyms Comment Organism
(Pl)EctB
-
Paenibacillus lautus
EctB
-
Paenibacillus lautus
More see also EC 2.6.1.19 Paenibacillus lautus

Temperature Optimum [┬░C]

Temperature Optimum [┬░C] Temperature Optimum Maximum [┬░C] Comment Organism
45
-
-
Paenibacillus lautus

Temperature Range [┬░C]

Temperature Minimum [┬░C] Temperature Maximum [┬░C] Comment Organism
10 60 activity range, profile overview Paenibacillus lautus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Paenibacillus lautus

pH Range

pH Minimum pH Maximum Comment Organism
5.5 10 40% of maximal activity within this range Paenibacillus lautus

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate PLP, residue Lys274 attaches the PLP cofactor to enzyme (Pl)EctB. The architecture of the PLP-binding site, formed by two monomeric subunits, overview Paenibacillus lautus

General Information

General Information Comment Organism
evolution EctB-type proteins are highly conserved Paenibacillus lautus
additional information structural homology modeling of (Pl)EctB using the crystal structure of the GABA transaminase from Arthrobacter aurescens (PDB ID 4ATP, EC 2.6.1.19) as template Paenibacillus lautus