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Literature summary extracted from

  • Burkhardt, I.; Ye, Z.; Janevska, S.; Tudzynski, B.; Dickschat, J.S.
    Biochemical and mechanistic characterization of the fungal reverse N-1-dimethylallyltryptophan synthase DMATS1Ff (2019), ACS Chem. Biol., 14, 2922-2931 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.5.1.B44 cloned into the expression vector pYE-express. The expression construct is transformed into Escherichia coli BL21 (DE3) Fusarium fujikuroi

Protein Variants

EC Number Protein Variants Comment Organism
2.5.1.B44 E109M the variant shows less than 1% of the wild-type Fusarium fujikuroi
2.5.1.B44 E90A completely inactive enzyme Fusarium fujikuroi
2.5.1.B44 F238R the variant shows less than 2% residual catalytic efficiency of the wild-type Fusarium fujikuroi
2.5.1.B44 I191Y the variant shows less than 2% residual catalytic efficiency of the wild-type Fusarium fujikuroi
2.5.1.B44 K187M the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi
2.5.1.B44 K253M the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi
2.5.1.B44 K332Q the variant shows 50% catalytic efficiency of the wild-type enzyme Fusarium fujikuroi
2.5.1.B44 R105K the variant exhibits 9% of the catalytic efficiency of the wild-type Fusarium fujikuroi
2.5.1.B44 R105M the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi
2.5.1.B44 R251K the variant shows 58% catalytic efficiency of the wild-type enzyme Fusarium fujikuroi
2.5.1.B44 R251M the variant shows 21% catalytic efficiency of the wild-type enzyme Fusarium fujikuroi
2.5.1.B44 R257L the variant shows less than 2% residual catalytic efficiency of the wild-type Fusarium fujikuroi
2.5.1.B44 S400Y the variant shows 56% catalytic efficiency of the wild-type enzyme Fusarium fujikuroi
2.5.1.B44 Y189F the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi
2.5.1.B44 Y255F the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi
2.5.1.B44 Y404F the variant shows diminished catalytic efficiency (kcat/Km) in comparison to the wild-type Fusarium fujikuroi

Inhibitors

EC Number Inhibitors Comment Organism Structure
2.5.1.B44 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 Fusarium fujikuroi
2.5.1.B44 Cu2+
-
 Fusarium fujikuroi
2.5.1.B44 geranyl diphosphate strong substrate inhibition above concentrations of 0.005 mM Fusarium fujikuroi
2.5.1.B44 Mn2+
-
 Fusarium fujikuroi
2.5.1.B44 Ni2+
-
 Fusarium fujikuroi
2.5.1.B44 Zn2+
-
 Fusarium fujikuroi

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.5.1.B44 0.0005
-
 geranyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.31
-
 L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.34
-
 dimethylallyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 3.4
-
 tyrosine pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 5.61
-
 5-hydroxy-L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 7.94
-
 D-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.5.1.B44 Ca2+ no dependency on divalent metal cations, but addition of Mg2+ and Ca2+ leads to slightly enhanced reaction rates Fusarium fujikuroi
2.5.1.B44 Mg2+ no dependency on divalent metal cations, but addition of Mg2+ and Ca2+ leads to slightly enhanced reaction rates Fusarium fujikuroi

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan Fusarium fujikuroi
-
 diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan Fusarium fujikuroi IMI58289
-
 diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.5.1.B44 Fusarium fujikuroi
-
-
-
2.5.1.B44 Fusarium fujikuroi IMI58289
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.5.1.B44
-
 Fusarium fujikuroi

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.5.1.B44 dimethylallyl diphosphate + 5-hydroxy-L-tryptophan
-
 Fusarium fujikuroi diphosphate + 5-hydroxy-1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + 5-hydroxy-L-tryptophan
-
 Fusarium fujikuroi IMI58289 diphosphate + 5-hydroxy-1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + D-tryptophan
-
 Fusarium fujikuroi diphosphate + 1-(2-methylbut-3-en-2-yl)-D-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + D-tryptophan
-
 Fusarium fujikuroi IMI58289 diphosphate + 1-(2-methylbut-3-en-2-yl)-D-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan
-
 Fusarium fujikuroi diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan the reaction catalyzed by the enzyme (DMATS1Ff) is nonstereospecific. No activity with isopentenyl diphosphate, farnesyl diphosphate and geranylgeranyl diphosphate Fusarium fujikuroi diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan
-
 Fusarium fujikuroi IMI58289 diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + L-tryptophan the reaction catalyzed by the enzyme (DMATS1Ff) is nonstereospecific. No activity with isopentenyl diphosphate, farnesyl diphosphate and geranylgeranyl diphosphate Fusarium fujikuroi IMI58289 diphosphate + 1-(2-methylbut-3-en-2-yl)-L-tryptophan
-
 ?
2.5.1.B44 dimethylallyl diphosphate + tyrosine
-
 Fusarium fujikuroi diphosphate + O-(3-methylbut-2-en-1-yl)-L-tyrosine
-
 ?
2.5.1.B44 dimethylallyl diphosphate + tyrosine
-
 Fusarium fujikuroi IMI58289 diphosphate + O-(3-methylbut-2-en-1-yl)-L-tyrosine
-
 ?
2.5.1.B44 geranyl diphosphate + L-tryptophan
-
 Fusarium fujikuroi diphosphate + 1-(3,7-dimethylocta-1,6-dien-3-yl)-L-tryptophan
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.5.1.B44 DMATS1Ff
-
 Fusarium fujikuroi

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.5.1.B44 35
-
-
 Fusarium fujikuroi

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
2.5.1.B44 25 40 25°C: about 50% of maximal activity, 40°C: about 70% of maximal activity Fusarium fujikuroi

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.5.1.B44 0.00084
-
 geranyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.0038
-
 D-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.0056
-
 tyrosine pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.02
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme K253M Fusarium fujikuroi
2.5.1.B44 0.02
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme K253M Fusarium fujikuroi
2.5.1.B44 0.03
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme Y189F Fusarium fujikuroi
2.5.1.B44 0.03
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme Y189F Fusarium fujikuroi
2.5.1.B44 0.06
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme Y255F Fusarium fujikuroi
2.5.1.B44 0.06
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme Y255F Fusarium fujikuroi
2.5.1.B44 0.1
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme I191Y Fusarium fujikuroi
2.5.1.B44 0.1
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme I191Y Fusarium fujikuroi
2.5.1.B44 0.1
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme R105K Fusarium fujikuroi
2.5.1.B44 0.1
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme R105K Fusarium fujikuroi
2.5.1.B44 0.11
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme F238R Fusarium fujikuroi
2.5.1.B44 0.11
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme F238R Fusarium fujikuroi
2.5.1.B44 0.12
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme R251M Fusarium fujikuroi
2.5.1.B44 0.12
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme R251M Fusarium fujikuroi
2.5.1.B44 0.16
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme E109M Fusarium fujikuroi
2.5.1.B44 0.16
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme E109M Fusarium fujikuroi
2.5.1.B44 0.16
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme R105M Fusarium fujikuroi
2.5.1.B44 0.16
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme R105M Fusarium fujikuroi
2.5.1.B44 0.21
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme K187M Fusarium fujikuroi
2.5.1.B44 0.21
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme K187M Fusarium fujikuroi
2.5.1.B44 0.44
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme R257L Fusarium fujikuroi
2.5.1.B44 0.44
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme R257L Fusarium fujikuroi
2.5.1.B44 0.69
-
 dimethylallyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.72
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme K332Q Fusarium fujikuroi
2.5.1.B44 0.72
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme K332Q Fusarium fujikuroi
2.5.1.B44 1.27
-
 L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 1.49
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme S400Y Fusarium fujikuroi
2.5.1.B44 1.49
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme S400Y Fusarium fujikuroi
2.5.1.B44 2.26
-
 5-hydroxy-L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 2.87
-
 L-tryptophan pH 7.5, 35°C, mutant enzyme R251K Fusarium fujikuroi
2.5.1.B44 2.87
-
 dimethylallyl diphosphate pH 7.5, 35°C, mutant enzyme R251K Fusarium fujikuroi

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.5.1.B44 8.5
-
-
 Fusarium fujikuroi

pH Range

EC Number pH Minimum pH Maximum Comment Organism
2.5.1.B44 6 9 pH 6.0: about 55% of maximal activity, pH 9.0: about 90% of maximal activity Fusarium fujikuroi

Ki Value [mM]

EC Number Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
2.5.1.B44 0.278
-
 geranyl diphosphate pH 7.5, 35°C Fusarium fujikuroi

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.5.1.B44 0.00048
-
 D-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.02
-
 tyrosine pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.16
-
 geranyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 0.4
-
 5-hydroxy-L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 1.99
-
 dimethylallyl diphosphate pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi
2.5.1.B44 4.07
-
 L-tryptophan pH 7.5, 35°C, wild-type enzyme Fusarium fujikuroi