PDBsum entry 5ixj

Lyase

PDB id: 5ixj

Contents

Protein chains

384 a.a.

Ligands

THR ×4

Metals

_NA ×4

Waters ×846

PDB id:

5ixj

Name:

Lyase

Title:

Tryptophan synthase beta-subunit from pyrococcus furiosus with l- threonine non-covalently bound in the active site

Structure:

Tryptophan synthase beta chain 1. Chain: a, b, c, d. Engineered: yes

Source:

Pyrococcus furiosus (strain atcc 43587 / dsm 3638 / jcm 8422 / vc1). Organism_taxid: 186497. Strain: atcc 43587 / dsm 3638 / jcm 8422 / vc1. Gene: trpb1, pf1706. Expressed in: escherichia coli. Expression_system_taxid: 562

Resolution:

1.54Å R-factor: 0.176 R-free: 0.195

Authors:

A.R.Buller,M.Herger,F.H.Arnold

Key ref:

M.Herger et al. (2016). Synthesis of β-Branched Tryptophan Analogues Using an Engineered Subunit of Tryptophan Synthase. J Am Chem Soc, 138, 8388-8391. PubMed id: 27355405 DOI: 10.1021/jacs.6b04836

Date:

23-Mar-16 Release date: 17-Aug-16

Protein chains

Q8U093  (TRPB1_PYRFU) -  Tryptophan synthase beta chain 1 from Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)

Seq: 388 a.a.

Struc: 384 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.4.2.1.20 - tryptophan synthase.
• Pathway: Tryptophan Biosynthesis
• Reaction: (1S,2R)-1-C-(indol-3-yl)glycerol 3-phosphate + L-serine = D-glyceraldehyde 3-phosphate + L-tryptophan + H2O

(1S,2R)-1-C-(indol-3-yl)glycerol 3-phosphate + L-serine (Bound ligand (Het Group name = THR) matches with 87.50% similarity) = D-glyceraldehyde 3-phosphate + L-tryptophan + H2O

• Cofactor: Pyridoxal 5'-phosphate

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

reference

DOI no: 10.1021/jacs.6b04836

J Am Chem Soc 138:8388-8391 (2016)

PubMed id: 27355405

Synthesis of β-Branched Tryptophan Analogues Using an Engineered Subunit of Tryptophan Synthase.

M.Herger, P.van Roye, D.K.Romney, S.Brinkmann-Chen, A.R.Buller, F.H.Arnold.

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.