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Literature summary for 4.1.99.2 extracted from

  • Zheng, R.C.; Tang, X.L.; Suo, H.; Feng, L.L.; Liu, X.; Yang, J.; Zheng, Y.G.
    Biochemical characterization of a novel tyrosine phenol-lyase from Fusobacterium nucleatum for highly efficient biosynthesis of L-DOPA (2018), Enzyme Microb. Technol., 112, 88-93 .
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
pyridoxal 5'-phosphate dependent on, activates Fusobacterium nucleatum subsp. nucleatum

Application

Application Comment Organism
synthesis high activity and high level expression of Fn-TPL in Escherichia coli provides a cheap and robust biocatalyst for large-scale production of L-DOPA Fusobacterium nucleatum subsp. nucleatum

Cloned(Commentary)

Cloned (Comment) Organism
gene tnaA, recombinant expression of codon-optimized His6-tagged enzyme in Escherichia coli strain BL21(DE3) Fusobacterium nucleatum subsp. nucleatum

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.18
-
L-tyrosine recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
3.6
-
phenol recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
6.28
-
L-Dopa recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
15.62
-
pyrocatechol recombinant enzyme, L-DOPA formation is measured from dicatechol, pyruvate, and ammonium, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-tyrosine + H2O Fusobacterium nucleatum subsp. nucleatum L-tyrosine is the physiological substrate of enzyme Fn-TPL phenol + pyruvate + NH3
-
r
L-tyrosine + H2O Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 L-tyrosine is the physiological substrate of enzyme Fn-TPL phenol + pyruvate + NH3
-
r

Organism

Organism UniProt Comment Textmining
Fusobacterium nucleatum subsp. nucleatum Q8RHM6
-
-
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 Q8RHM6
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His6-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and dialysis Fusobacterium nucleatum subsp. nucleatum

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-cysteine + H2O 12% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum thiophenol + pyruvate + NH3
-
r
L-DOPA + H2O 42% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum pyrocatechol + pyruvate + NH4+
-
r
L-phenylalanine + H2O 7% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum ? + pyruvate + NH3
-
r
L-serine + H2O 9% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum pyruvate + NH3
-
r
L-tryptophan + H2O 8% of the activity with L-tyrosine, reaction of EC 4.1.99.1 Fusobacterium nucleatum subsp. nucleatum indole + pyruvate + NH3
-
r
L-tryptophan + H2O 8% of the activity with L-tyrosine, reaction of EC 4.1.99.1 Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 indole + pyruvate + NH3
-
r
L-tyrosine + H2O L-tyrosine is the physiological substrate of enzyme Fn-TPL Fusobacterium nucleatum subsp. nucleatum phenol + pyruvate + NH3
-
r
L-tyrosine + H2O alpha,beta-elimination of L-tyrosine, best substrate Fusobacterium nucleatum subsp. nucleatum phenol + pyruvate + NH3
-
r
L-tyrosine + H2O L-tyrosine is the physiological substrate of enzyme Fn-TPL Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 phenol + pyruvate + NH3
-
r
L-tyrosine + H2O alpha,beta-elimination of L-tyrosine, best substrate Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 phenol + pyruvate + NH3
-
r
L-valine + H2O 6% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum ? + pyruvate + NH3
-
r
additional information low activity of Fn-TPL is observed toward natural amino acids including L-serine, L-cysteine, L-phenylalanine, L-valine, L-tryptophan, no activity with L-threonine, L-glutamine, L-alanine, D-alanine, L-arginine, L-leucine, L-histidine, L-aspartic acid, L-methionine, and D-methionine Fusobacterium nucleatum subsp. nucleatum ?
-
?
additional information low activity of Fn-TPL is observed toward natural amino acids including L-serine, L-cysteine, L-phenylalanine, L-valine, L-tryptophan, no activity with L-threonine, L-glutamine, L-alanine, D-alanine, L-arginine, L-leucine, L-histidine, L-aspartic acid, L-methionine, and D-methionine Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 ?
-
?
phenol + pyruvate + NH3
-
Fusobacterium nucleatum subsp. nucleatum L-tyrosine + H2O
-
r
pyrocatechol + pyruvate + NH4+
-
Fusobacterium nucleatum subsp. nucleatum L-DOPA + H2O
-
r
pyrocatechol + pyruvate + NH4+
-
Fusobacterium nucleatum subsp. nucleatum ATCC 25586 / CIP 101130 / JCM 8532 / LMG 13131 L-DOPA + H2O
-
r
S-methyl-L-cysteine + H2O 13% of the activity with L-tyrosine Fusobacterium nucleatum subsp. nucleatum methanthiol + pyruvate + NH3
-
r

Synonyms

Synonyms Comment Organism
Fn-TPL
-
Fusobacterium nucleatum subsp. nucleatum
TnaA
-
Fusobacterium nucleatum subsp. nucleatum
TPL
-
Fusobacterium nucleatum subsp. nucleatum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
20
-
assay at Fusobacterium nucleatum subsp. nucleatum

Temperature Range [°C]

Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
20 80 activity range, profile overview Fusobacterium nucleatum subsp. nucleatum

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.44
-
phenol recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
0.54
-
L-Dopa recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
1.72
-
L-tyrosine recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
2.68
-
pyrocatechol recombinant enzyme, L-DOPA formation is measured from dicatechol, pyruvate, and ammonium, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
assay at Fusobacterium nucleatum subsp. nucleatum

pH Range

pH Minimum pH Maximum Comment Organism
5.5 10 activity range, profile overview Fusobacterium nucleatum subsp. nucleatum

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on, activates, Km value is 0.0034 mM Fusobacterium nucleatum subsp. nucleatum

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.09
-
L-Dopa recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
0.12
-
phenol recombinant enzyme, L-tyrosine formation is measured from phenol, pyruvate, and ammonium, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum
9.58
-
L-tyrosine recombinant enzyme, pH 8.5, 20°C Fusobacterium nucleatum subsp. nucleatum