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Direct Enzymatic Synthesis of a Deep-Blue Fluorescent Noncanonical Amino Acid from Azulene and Serine.

ABSTRACT: We report a simple, one-step enzymatic synthesis of the blue fluorescent noncanonical amino acid ?-(1-azulenyl)-l-alanine (AzAla). By using an engineered tryptophan synthase ?-subunit (TrpB), stereochemically pure AzAla can be synthesized at scale starting from commercially available azulene and l-serine. Mutation of a universally conserved catalytic glutamate in the active site to glycine has only a modest effect on native activity with indole but abolishes activity on azulene, suggesting that this glutamate activates azulene for nucleophilic attack by stabilization of the aromatic ion.

SUBMITTER: Watkins EJ

PROVIDER: S-EPMC6980211 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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Direct Enzymatic Synthesis of a Deep-Blue Fluorescent Noncanonical Amino Acid from Azulene and Serine.

Watkins Ella J EJ Almhjell Patrick J PJ Arnold Frances H FH

Chembiochem : a European journal of chemical biology 20191118 1-2

We report a simple, one-step enzymatic synthesis of the blue fluorescent noncanonical amino acid β-(1-azulenyl)-l-alanine (AzAla). By using an engineered tryptophan synthase β-subunit (TrpB), stereochemically pure AzAla can be synthesized at scale starting from commercially available azulene and l-serine. Mutation of a universally conserved catalytic glutamate in the active site to glycine has only a modest effect on native activity with indole but abolishes activity on azulene, suggesting that ...[more]

PMID: 31513332

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