PDBsum entry 5oy2

Oxidoreductase

PDB id: 5oy2

Contents

Protein chain

327 a.a.

Ligands

HEM

NAG ×5

PO4

3DM

GOL ×3

Metals

_MG

_CL

Waters ×368

PDB id:

5oy2

Name:

Oxidoreductase

Title:

Direct-evolutioned unspecific peroxygenase from agrocybe aegerita, in complex with dmp

Structure:

Unspecific peroxygenase. Chain: a. Synonym: upo. Engineered: yes. Mutation: yes

Source:

Agrocybe aegerita. Black poplar mushroom. Organism_taxid: 5400. Gene: apo1. Expressed in: komagataella phaffii cbs 7435. Expression_system_taxid: 981350. Expression_system_variant: x-33. Expression_system_atcc_number: 76273.

Resolution:

1.36Å R-factor: 0.160 R-free: 0.170

Authors:

M.Ramirez-Escudero,J.Sanz-Aparicio

Key ref:

M.Ramirez-Escudero et al. (2018). Structural Insights into the Substrate Promiscuity of a Laboratory-Evolved Peroxygenase. ACS Chem Biol, 13, 3259-3268. PubMed id: 30376293 DOI: 10.1021/acschembio.8b00500

Date:

07-Sep-17 Release date: 17-Apr-19

Protein chain

B9W4V6  (APO1_CYCAE) -  Aromatic peroxygenase from Cyclocybe aegerita

Seq: 371 a.a.

Struc: 327 a.a.*

* PDB and UniProt seqs differ at 5 residue positions (black crosses)

Enzyme reactions

• Enzyme class: E.C.1.11.2.1 - unspecific peroxygenase.
• Reaction: RH + H2O2 = ROH + H2O
• Cofactor: Heme-thiolate

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

Added reference

DOI no: 10.1021/acschembio.8b00500

ACS Chem Biol 13:3259-3268 (2018)

PubMed id: 30376293

Structural Insights into the Substrate Promiscuity of a Laboratory-Evolved Peroxygenase.

M.Ramirez-Escudero, P.Molina-Espeja, P.Gomez de Santos, M.Hofrichter, J.Sanz-Aparicio, M.Alcalde.

ABSTRACT

Because of their minimal requirements, substrate promiscuity and product selectivity, fungal peroxygenases are now considered to be the jewel in the crown of C-H oxyfunctionalization biocatalysts. In this work, the crystal structure of the first laboratory-evolved peroxygenase expressed by yeast was determined at a resolution of 1.5 Å. Notable differences were detected between the evolved and native peroxygenase from Agrocybe aegerita, including the presence of a full N-terminus and a broader heme access channel due to the mutations that accumulated through directed evolution. Further mutagenesis and soaking experiments with a palette of peroxygenative and peroxidative substrates suggested dynamic trafficking through the heme channel as the main driving force for the exceptional substrate promiscuity of peroxygenase. Accordingly, this study provides the first structural evidence at an atomic level regarding the mode of substrate binding for this versatile biocatalyst, which is discussed within a biological and chemical context.