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Literature summary for 4.1.99.2 extracted from

  • Kim, S.; Sung, B.H.; Kim, S.C.; Lee, H.S.
    Genetic incorporation of l-dihydroxyphenylalanine (DOPA) biosynthesized by a tyrosine phenol-lyase (2018), Chem. Commun. (Camb.), 54, 3002-3005 .
    View publication on PubMed

Application

Application Comment Organism
synthesis the tyrosine phenol lyase (TPL) catalyzes synthesis of L-DOPA and can be used for large scale L-DOPA synthesis with direct incorporation of L-DOPA into proteins Citrobacter freundii

Cloned(Commentary)

Cloned (Comment) Organism
gene tnaA, recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3), subcloning in Escherichia coli strain DH10beta Citrobacter freundii

Protein Variants

Protein Variants Comment Organism
additional information L-dihydroxyphenylalanine (DOPA) is biosynthesized by a tyrosine-phenol lyase from catechol, pyruvate, and ammonia in recombinant Escherichia coli expressing the enzyme, and the biosynthesized amino acid is directly incorporated into proteins. Three biochemical experiments with mutant proteins containing DOPA confirm the genetic incorporation of biosynthesized DOPA, and reveal its potential for various biochemical applications Citrobacter freundii

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-tyrosine + H2O Citrobacter freundii
-
phenol + pyruvate + NH3
-
r
L-tyrosine + H2O Citrobacter freundii ATCC 8090
-
phenol + pyruvate + NH3
-
r

Organism

Organism UniProt Comment Textmining
Citrobacter freundii P31013
-
-
Citrobacter freundii ATCC 8090 P31013
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography Citrobacter freundii

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
catechol + pyruvate
-
Citrobacter freundii L-DOPA + H2O
-
r
catechol + pyruvate
-
Citrobacter freundii ATCC 8090 L-DOPA + H2O
-
r
L-tyrosine + H2O
-
Citrobacter freundii phenol + pyruvate + NH3
-
r
L-tyrosine + H2O
-
Citrobacter freundii ATCC 8090 phenol + pyruvate + NH3
-
r

Synonyms

Synonyms Comment Organism
TPL
-
Citrobacter freundii

Cofactor

Cofactor Comment Organism Structure
pyridoxal 5'-phosphate dependent on Citrobacter freundii