Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Ohshida, T.; Koba, K.; Hayashi, J.; Yoneda, K.; Ohmori, T.; Ohshima, T.; Sakuraba, H.
    A novel bifunctional aspartate kinase-homoserine dehydrogenase from the hyperthermophilic bacterium, Thermotoga maritima (2018), Biosci. Biotechnol. Biochem., 82, 2084-2093 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.3 gene TM_0547 , sequence comparisons, recombinant expression of the enzyme in Escherichia coli strain BL21(DE3) Thermotoga maritima
2.7.2.4 expressed in Escherichia coli BL21(DE3) Codon plus-RIPL cells Thermotoga maritima
2.7.2.4 gene TM_0547 , sequence comparisons, recombinant expression of the enzyme in Escherichia coli strain BL21(DE3) Thermotoga maritima

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.3 additional information L-homoserine oxidation of the Thermotoga maritima enzyme is almost impervious to inhibition by L-threonine, while L-threonine inhibits AK activity in a cooperative manner. The distinctive sequence of the regulatory domain in Thermotoga maritima AK-HseDH is likely responsible for the unique sensitivity to L-threonine. The quaternary structure of this enzyme is not affected by L-threonine Thermotoga maritima
2.7.2.4 L-threonine inhibits AK activity in a cooperative manner by 90% at 0.35 mM. The distinctive sequence of the regulatory domain in Thermotoga maritima AK-HseDH is likely responsible for the unique sensitivity to L-threonine. The quaternary structure of this enzyme is not affected by L-threonine Thermotoga maritima
2.7.2.4 additional information L-homoserine oxidation of the Thermotoga maritima enzyme is almost impervious to inhibition by L-threonine Thermotoga maritima

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.3 0.064
-
 NADP+ recombinant enzyme, pH 10.5, 55°C Thermotoga maritima
1.1.1.3 1.81
-
 L-homoserine recombinant enzyme, pH 10.5, 55°C Thermotoga maritima
2.7.2.4 3.54
-
 ATP at pH 7.0 and 55°C Thermotoga maritima
2.7.2.4 3.54
-
 ATP recombinant enzyme, pH 7.5, 55°C Thermotoga maritima
2.7.2.4 4.66
-
 L-aspartate at pH 7.0 and 55°C Thermotoga maritima
2.7.2.4 4.66
-
 L-aspartate recombinant enzyme, pH 7.5, 55°C Thermotoga maritima

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.2.4 Mg2+ 4 mM used in assay conditions Thermotoga maritima

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.3 450000
-
 about, recombinant enzyme, gel filtration Thermotoga maritima
2.7.2.4 450000
-
 gel filtration Thermotoga maritima
2.7.2.4 450000
-
 about, recombinant enzyme, gel filtration Thermotoga maritima

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.3 L-homoserine + NADP+ Thermotoga maritima
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+ Thermotoga maritima DSM 3109
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+ Thermotoga maritima ATCC 43589
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+ Thermotoga maritima JCM 10099
-
 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
2.7.2.4 ATP + L-aspartate Thermotoga maritima
-
 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate Thermotoga maritima DSM 3109
-
 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate Thermotoga maritima ATCC 43589
-
 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate Thermotoga maritima JCM 10099
-
 ADP + 4-phospho-L-aspartate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.3 Thermotoga maritima Q9WZ17
-
-
1.1.1.3 Thermotoga maritima ATCC 43589 Q9WZ17
-
-
1.1.1.3 Thermotoga maritima DSM 3109 Q9WZ17
-
-
1.1.1.3 Thermotoga maritima JCM 10099 Q9WZ17
-
-
2.7.2.4 Thermotoga maritima
-
-
-
2.7.2.4 Thermotoga maritima Q9WZ17
-
-
2.7.2.4 Thermotoga maritima ATCC 43589 Q9WZ17
-
-
2.7.2.4 Thermotoga maritima DSM 3109 Q9WZ17
-
-
2.7.2.4 Thermotoga maritima JCM 10099 Q9WZ17
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.3 recombinant enzyme from Escherichia coli strain BL21(DE3) by heat treatment, hydrophobic interaction chromatography, gel filtration, hydroxyapatite chromatography, and ultrafiltration Thermotoga maritima
2.7.2.4 recombinant enzyme from Escherichia coli strain BL21(DE3) by heat treatment, hydrophobic interaction chromatography, gel filtration, hydroxyapatite chromatography, and ultrafiltration Thermotoga maritima
2.7.2.4 Toyopearl ether-650M column chromatography, Superdex 200 gel filtration and Cellulofine HAP hydroxyl apatite column chromatography Thermotoga maritima

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.1.1.3 66.5
-
 purified recombinant enzyme, pH 10.5, 55°C Thermotoga maritima
2.7.2.4 103
-
 purified recombinant enzyme, pH 7.5, 55°C Thermotoga maritima

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.3 L-homoserine + NADP+
-
 Thermotoga maritima L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+
-
 Thermotoga maritima DSM 3109 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+
-
 Thermotoga maritima ATCC 43589 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 L-homoserine + NADP+
-
 Thermotoga maritima JCM 10099 L-aspartate 4-semialdehyde + NADPH + H+
-
 ?
1.1.1.3 additional information the bifunctional enzyme also exhibits aspartokinase (AK) activity, EC 2.7.2.4. It shows substantial activities of both AK and homoserine dehydrogenase (HseDH) Thermotoga maritima ?
-
-
1.1.1.3 additional information the bifunctional enzyme also exhibits aspartokinase (AK) activity, EC 2.7.2.4. It shows substantial activities of both AK and homoserine dehydrogenase (HseDH) Thermotoga maritima DSM 3109 ?
-
-
1.1.1.3 additional information the bifunctional enzyme also exhibits aspartokinase (AK) activity, EC 2.7.2.4. It shows substantial activities of both AK and homoserine dehydrogenase (HseDH) Thermotoga maritima ATCC 43589 ?
-
-
1.1.1.3 additional information the bifunctional enzyme also exhibits aspartokinase (AK) activity, EC 2.7.2.4. It shows substantial activities of both AK and homoserine dehydrogenase (HseDH) Thermotoga maritima JCM 10099 ?
-
-
2.7.2.4 ATP + L-aspartate
-
 Thermotoga maritima ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate
-
 Thermotoga maritima DSM 3109 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate
-
 Thermotoga maritima ATCC 43589 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 ATP + L-aspartate
-
 Thermotoga maritima JCM 10099 ADP + 4-phospho-L-aspartate
-
 ?
2.7.2.4 additional information the bifunctional enzyme also exhibits L-homoserine dehydrogenase activity, EC 1.1.1.3. It shows substantial activities of both aspartate kinase (AK) and homoserine dehydrogenase (HseDH) Thermotoga maritima ?
-
-
2.7.2.4 additional information the bifunctional enzyme also exhibits L-homoserine dehydrogenase activity, EC 1.1.1.3. It shows substantial activities of both aspartate kinase (AK) and homoserine dehydrogenase (HseDH) Thermotoga maritima DSM 3109 ?
-
-
2.7.2.4 additional information the bifunctional enzyme also exhibits L-homoserine dehydrogenase activity, EC 1.1.1.3. It shows substantial activities of both aspartate kinase (AK) and homoserine dehydrogenase (HseDH) Thermotoga maritima ATCC 43589 ?
-
-
2.7.2.4 additional information the bifunctional enzyme also exhibits L-homoserine dehydrogenase activity, EC 1.1.1.3. It shows substantial activities of both aspartate kinase (AK) and homoserine dehydrogenase (HseDH) Thermotoga maritima JCM 10099 ?
-
-

Subunits

EC Number Subunits Comment Organism
1.1.1.3 homopentamer or homohexamer x * 81000, recombinant enzyme, SDS-PAGE, x * 81433, sequence calculation Thermotoga maritima
2.7.2.4 homopentamer or homohexamer 5 or 6 * 81000, SDS-PAGE Thermotoga maritima
2.7.2.4 homopentamer or homohexamer 5 or 6 * 81433, calculated from amino acid sequence Thermotoga maritima
2.7.2.4 homopentamer or homohexamer x * 81000, recombinant enzyme, SDS-PAGE, x * 81433, sequence calculation Thermotoga maritima

Synonyms

EC Number Synonyms Comment Organism
1.1.1.3 AK-HseDH
-
 Thermotoga maritima
1.1.1.3 bifunctional aspartate kinase-homoserine dehydrogenase
-
 Thermotoga maritima
1.1.1.3 HseDH
-
 Thermotoga maritima
1.1.1.3 More see also EC 2.7.2.4 Thermotoga maritima
1.1.1.3 TM_0547
-
 Thermotoga maritima
2.7.2.4 AK-HseDH
-
 Thermotoga maritima
2.7.2.4 AK-HseDH bifunctional enzyme with aspartate kinase and homoserine dehydrogenase activities Thermotoga maritima
2.7.2.4 bifunctional aspartate kinase-homoserine dehydrogenase
-
 Thermotoga maritima
2.7.2.4 HseDH
-
 Thermotoga maritima
2.7.2.4 More see also EC 1.1.1.3 Thermotoga maritima
2.7.2.4 TM_0547
-
 Thermotoga maritima

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.3 70
-
 Hse oxidation Thermotoga maritima
2.7.2.4 70
-
-
 Thermotoga maritima
2.7.2.4 70
-
 L-aspartate phosphorylation Thermotoga maritima

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
1.1.1.3 50 80 over 30% of maximal activity within this range, profile overview Thermotoga maritima
2.7.2.4 50 85 over 50% of maximal activity at 50°C, 80% at 85°C, profile overview Thermotoga maritima

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.3 50 100 purified recombinant enzyme, 10 min, completely stable at 50-90°C, loss of 40% activity at 100°C Thermotoga maritima
2.7.2.4 50 100 purified recombinant enzyme, 10 min, completely stable at 50-90°C, loss of 40% activity at 100°C Thermotoga maritima
2.7.2.4 50 90 the enzyme remains stable (more than 90% activity) after 10 min incubation between 50 and 90°C and drops to about 50% activity after 10 min at 100°C Thermotoga maritima

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.3 10.5
-
 Hse oxidation Thermotoga maritima
2.7.2.4 7 8 L-aspartate phosphorylation Thermotoga maritima
2.7.2.4 7.5
-
-
 Thermotoga maritima

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.1.1.3 6.5 11 over 60% of maximal activity within this range, profile overview Thermotoga maritima
2.7.2.4 6.5 9.5 over 60% of maximal activity within this range, profile overview Thermotoga maritima

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
1.1.1.3 5 12 purified recombinant enzyme, 10 min, 50°C, completely stable at pH 5.5-10.5, loss of 25% activity at pH 11.0, loss of 70% activity at pH 5.0 and pH 11.5, inactivation at pH 12.0 Thermotoga maritima
2.7.2.4 5 12 purified recombinant enzyme, 10 min, 50°C, completely stable at pH 5.5-10.5, loss of 25% activity at pH 11.0, loss of 70% activity at pH 5.0 and pH 11.5, inactivation at pH 12.0 Thermotoga maritima
2.7.2.4 5.5 10.5 the enzyme loses no activity when incubated at pH values between 5.5 and 10.5 for 10 min at 50°C Thermotoga maritima

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.3 NADP+
-
 Thermotoga maritima
1.1.1.3 NADPH
-
 Thermotoga maritima
2.7.2.4 NADP+
-
 Thermotoga maritima
2.7.2.4 NADPH
-
 Thermotoga maritima

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
2.7.2.4 0.037
-
 pH 7.5, 55°C Thermotoga maritima L-threonine

General Information

EC Number General Information Comment Organism
1.1.1.3 evolution the orientation of the three domains in the bifunctional aspartate kinase-homoserine dehydrogenase (AK-HseDH) homologue found in Thermotoga maritima totally differs from those observed in previously known AK-HseDHs, the domains line up in the order HseDH, AK, and regulatory domain Thermotoga maritima
1.1.1.3 metabolism biosynthetic pathway from L-aspartate to L-homoserine involving the bifunctional enzyme, overview Thermotoga maritima
1.1.1.3 physiological function aspartate kinase (AK, EC 2.7.2.4) and homoserine dehydrogenase (HseDH) are involved in the biosynthetic pathway from L-aspartate to L-homoserine (Hse) in plants and microorganisms. Hse is a common precursor for the synthesis of L-methionine, L-threonine, and L-isoleucine. At the first step in this pathway, L-aspartate is phosphorylated to beta-aspartyl phosphate (beta-Ap) by AK Thermotoga maritima
2.7.2.4 evolution the orientation of the three domains in the bifunctional aspartate kinase-homoserine dehydrogenase (AK-HseDH) homologue found in Thermotoga maritima totally differs from those observed in previously known AK-HseDHs, the domains line up in the order HseDH, AK, and regulatory domain Thermotoga maritima
2.7.2.4 metabolism biosynthetic pathway from L-aspartate to L-homoserine involving the bifunctional enzyme, overview Thermotoga maritima
2.7.2.4 physiological function aspartate kinase (AK) and homoserine dehydrogenase (HseDH, EC 1.1.1.3) are involved in the biosynthetic pathway from L-aspartate to L-homoserine (Hse) in plants and microorganisms. Hse is a common precursor for the synthesis of L-methionine, L-threonine, and L-isoleucine. At the first step in this pathway, L-aspartate is phosphorylated to beta-aspartyl phosphate (beta-Ap) by AK Thermotoga maritima