6CUV

Engineered Holo TrpB from Pyrococcus furiosus, PfTrpB7E6

Summary for 6CUV

• Entry DOI: 10.2210/pdb6cuv/pdb
• Descriptor: Tryptophan synthase beta chain 1, SODIUM ION, PHOSPHATE ION, ... (4 entities in total)
• Functional Keywords: internal aldimine, lyase
• Biological source: Pyrococcus furiosus (strain ATCC 43587 / DSM 3638 / JCM 8422 / Vc1)
• Total number of polymer chains: 4
• Total formula weight: 170735.14

Authors

Scheele, R.A.,Buller, A.R.,Boville, C.E.,Arnold, F.H. (deposition date: 2018-03-26, release date: 2018-09-26, Last modification date: 2023-11-15)

Primary citation

Boville, C.E.,Scheele, R.A.,Koch, P.,Brinkmann-Chen, S.,Buller, A.R.,Arnold, F.H.
Engineered Biosynthesis of beta-Alkyl Tryptophan Analogues.
Angew. Chem. Int. Ed. Engl., 57:14764-14768, 2018

PubMed Abstract: Noncanonical amino acids (ncAAs) with dual stereocenters at the α and β positions are valuable precursors to natural products and therapeutics. Despite the potential applications of such bioactive β-branched ncAAs, their availability is limited due to the inefficiency of the multistep methods used to prepare them. Herein we report a stereoselective biocatalytic synthesis of β-branched tryptophan analogues using an engineered variant of Pyrococcus furiosus tryptophan synthase (PfTrpB), PfTrpB . PfTrpB is the first biocatalyst to synthesize bulky β-branched tryptophan analogues in a single step, with demonstrated access to 27 ncAAs. The molecular basis for the efficient catalysis and broad substrate tolerance of PfTrpB was explored through X-ray crystallography and UV/Vis spectroscopy, which revealed that a combination of active-site and remote mutations increase the abundance and persistence of a key reactive intermediate. PfTrpB provides an operationally simple and environmentally benign platform for the preparation of β-branched tryptophan building blocks.

PubMed: 30215880
DOI: 10.1002/anie.201807998

Experimental method

X-RAY DIFFRACTION (2.26 Å)