PDBsum entry 5ixj

Lyase

PDB id: 5ixj

Contents

Protein chains

384 a.a.

Ligands

THR ×4

Metals

_NA ×4

Waters ×846

References listed in PDB file

Key reference

• Title: Synthesis of β-Branched tryptophan analogues using an engineered subunit of tryptophan synthase.
• Authors: M.Herger, P.Van roye, D.K.Romney, S.Brinkmann-Chen, A.R.Buller, F.H.Arnold.
• Ref.: J Am Chem Soc, 2016, 138, 8388-8391. [DOI no: 10.1021/jacs.6b04836]
• PubMed id: 27355405

Abstract

We report that l-threonine may substitute for l-serine in the β-substitution reaction of an engineered subunit of tryptophan synthase from Pyrococcus furiosus, yielding (2S,3S)-β-methyltryptophan (β-MeTrp) in a single step. The trace activity of the wild-type β-subunit on this substrate was enhanced more than 1000-fold by directed evolution. Structural and spectroscopic data indicate that this increase is correlated with stabilization of the electrophilic aminoacrylate intermediate. The engineered biocatalyst also reacts with a variety of indole analogues and thiophenol for diastereoselective C-C, C-N, and C-S bond-forming reactions. This new activity circumvents the 3-enzyme pathway that produces β-MeTrp in nature and offers a simple and expandable route to preparing derivatives of this valuable building block.