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

BRENDA support

Literature summary for 6.3.4.24 extracted from

  • Wang, Y.; Xu, H.; Harich, K.C.; White, R.H.
    Identification and characterization of a tyramine-glutamate ligase (MfnD) involved in methanofuran biosynthesis (2014), Biochemistry, 53, 6220-6230.
    View publication on PubMed
Search for more literature for 6.3.4.24

Crystallization (Commentary)

Crystallization (Comment) Organism
-
 Methanocaldococcus fervens

Protein Variants

Protein Variants Comment Organism
E21Q mutation causes a 3fold decrease in kcat(app) and a slight increase in KM(glutamate). The apparent second-order constants kcat/KM is comparable to the wild type Methanocaldococcus fervens
R251A mutation causes a 100fold decrease in kcat(app) and a more than 20fold increase in KM(glutamate), indicating that it plays an important role in catalysis Methanocaldococcus fervens
T253V mutation causes a 3fold decrease in kcat(app). The apparent second-order constants kcat/KM is comparable to the wild type Methanocaldococcus fervens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.6
-
 tyramine pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
0.8
-
 tyramine pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
0.8
-
 tyramine pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
1.1
-
 tyramine pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
1.5
-
 ATP pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
2
-
 ATP pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
2.3
-
 L-glutamate pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
2.5
-
 ATP pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
3
-
 ATP pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
3.6
-
 L-glutamate pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
7.3
-
 L-glutamate pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ the enzyme requires a divalent cation for activity. Maximum activity (100%) is observed with the addition of 10 mM Mn2+, followed by 89%, 56%, 46%, and 46% of relative activity by addition of 10 mM Mg2+, Co2+, Zn2+, and Fe2+, respectively. Less than 5% relative activities are observed when 10 mM Ni2+ or Ca2+ is added. However, no activity is detected by addition of Cu2+ or K+ Methanocaldococcus fervens
Fe2+ the enzyme requires a divalent cation for activity. Maximum activity (100%) is observed with the addition of 10 mM Mn2+, followed by 89%, 56%, 46%, and 46% of relative activity by addition of 10 mM Mg2+, Co2+, Zn2+, and Fe2+, respectively. Less than 5% relative activities are observed when 10 mM Ni2+ or Ca2+ is added. However, no activity is detected by addition of Cu2+ or K+ Methanocaldococcus fervens
Mg2+ the enzyme requires a divalent cation for activity. Maximum activity (100%) is observed with the addition of 10 mM Mn2+, followed by 89%, 56%, 46%, and 46% of relative activity by addition of 10 mM Mg2+, Co2+, Zn2+, and Fe2+, respectively. Less than 5% relative activities are observed when 10 mM Ni2+ or Ca2+ is added. However, no activity is detected by addition of Cu2+ or K+ Methanocaldococcus fervens
Mn2+ the enzyme requires a divalent cation for activity. Maximum activity (100%) is observed with the addition of 10 mM Mn2+, followed by 89%, 56%, 46%, and 46% of relative activity by addition of 10 mM Mg2+, Co2+, Zn2+, and Fe2+, respectively. Less than 5% relative activities are observed when 10 mM Ni2+ or Ca2+ is added. However, no activity is detected by addition of Cu2+ or K+ Methanocaldococcus fervens
Zn2+ the enzyme requires a divalent cation for activity. Maximum activity (100%) is observed with the addition of 10 mM Mn2+, followed by 89%, 56%, 46%, and 46% of relative activity by addition of 10 mM Mg2+, Co2+, Zn2+, and Fe2+, respectively. Less than 5% relative activities are observed when 10 mM Ni2+ or Ca2+ is added. However, no activity is detected by addition of Cu2+ or K+ Methanocaldococcus fervens

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
33000
-
 x * 33000, SDS-PAGE Methanocaldococcus fervens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + tyramine + L-glutamate Methanocaldococcus fervens the enzyme participates in the biosynthesis of methanofuran, a coenzymes involved in the reduction of carbon dioxide to methane ADP + phosphate + gamma-glutamyltyramine
-
 ?
ATP + tyramine + L-glutamate Methanocaldococcus fervens DSM 4213 the enzyme participates in the biosynthesis of methanofuran, a coenzymes involved in the reduction of carbon dioxide to methane ADP + phosphate + gamma-glutamyltyramine
-
 ?

Organism

Organism UniProt Comment Textmining
Methanocaldococcus fervens C7P8V7
-
-
Methanocaldococcus fervens DSM 4213 C7P8V7
-
-

Purification (Commentary)

Purification (Comment) Organism
expression in Escherichia coli Methanocaldococcus fervens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + tyramine + L-glutamate the enzyme participates in the biosynthesis of methanofuran, a coenzymes involved in the reduction of carbon dioxide to methane Methanocaldococcus fervens ADP + phosphate + gamma-glutamyltyramine
-
 ?
ATP + tyramine + L-glutamate maximum activity with ATP (100%). Activities with UTP (<0.2%), CTP (<0.08%), and GTP (<0.04%) are much lower. Reactions using tyrosine or phenylalanine instead of tyramine show none of the expected products. Enzymatic reactions including gamma-glutamylglutamate, pyroglutamate, or N-acetyl-glutamate instead of L-glutamate show no expected products. No activity with furyltyramine as substrate. Arg251 is an important residue for both catalysis and glutamate binding Methanocaldococcus fervens ADP + phosphate + gamma-glutamyltyramine
-
 ?
ATP + tyramine + L-glutamate the enzyme participates in the biosynthesis of methanofuran, a coenzymes involved in the reduction of carbon dioxide to methane Methanocaldococcus fervens DSM 4213 ADP + phosphate + gamma-glutamyltyramine
-
 ?
ATP + tyramine + L-glutamate maximum activity with ATP (100%). Activities with UTP (<0.2%), CTP (<0.08%), and GTP (<0.04%) are much lower. Reactions using tyrosine or phenylalanine instead of tyramine show none of the expected products. Enzymatic reactions including gamma-glutamylglutamate, pyroglutamate, or N-acetyl-glutamate instead of L-glutamate show no expected products. No activity with furyltyramine as substrate. Arg251 is an important residue for both catalysis and glutamate binding Methanocaldococcus fervens DSM 4213 ADP + phosphate + gamma-glutamyltyramine
-
 ?

Subunits

Subunits Comment Organism
? x * 33000, SDS-PAGE Methanocaldococcus fervens

Synonyms

Synonyms Comment Organism
Mefer_1180
-
 Methanocaldococcus fervens
mfnD
-
 Methanocaldococcus fervens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
70
-
 assay at Methanocaldococcus fervens

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
80
-
 wild-type MfnD and its three variants (E21Q, R251A and T253V). No protein aggregation and precipitation is observed when proteins are incubated for 10 min at 80 °C in 25 mM Tricine/CAPS/TES buffer at pH 10.5 Methanocaldococcus fervens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.05
-
 tyramine pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
0.07
-
 ATP pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
1.3
-
 L-glutamate pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
1.4
-
 ATP pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
1.4
-
 tyramine pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
1.7
-
 tyramine pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
2.1
-
 ATP pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
2.7
-
 L-glutamate pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
4.8
-
 tyramine pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
5.8
-
 L-glutamate pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
5.9
-
 ATP pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.7
-
 broad pH optimum with a maximum around pH 6.7 Methanocaldococcus fervens

pH Range

pH Minimum pH Maximum Comment Organism
6.3 10.5 pH 6.3: about 80% of maximal activity, pH 10.5: about 60% of maximal activity Methanocaldococcus fervens

General Information

General Information Comment Organism
physiological function the enzyme participates in the biosynthesis of methanofuran, a coenzymes involved in the reduction of carbon dioxide to methane Methanocaldococcus fervens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.02
-
 ATP pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
0.06
-
 tyramine pH 6.7, 70°C, mutant enzyme R251A Methanocaldococcus fervens
0.36
-
 L-glutamate pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
0.37
-
 L-glutamate pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
0.56
-
 ATP pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
1.1
-
 ATP pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
1.7
-
 tyramine pH 6.7, 70°C, mutant enzyme E21Q Methanocaldococcus fervens
2.3
-
 tyramine pH 6.7, 70°C, mutant enzyme T253V Methanocaldococcus fervens
2.5
-
 L-glutamate pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
3.9
-
 ATP pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens
6
-
 tyramine pH 6.7, 70°C, wild-type enzyme Methanocaldococcus fervens