Reconstitution of full-length P450BM3 with an artificial metal complex by utilising the transpeptidase Sortase A

Keita Omura; Yuichiro Aiba; Hiroki Onoda; Joshua Kyle Stanfield; Shinya Ariyasu; Hiroshi Sugimoto; Yoshitsugu Shiro; Osami Shoji; Yoshihito Watanabe

doi:10.1039/c8cc02760a

Reconstitution of full-length P450BM3 with an artificial metal complex by utilising the transpeptidase Sortase A

Chem Commun (Camb). 2018 Jul 12;54(57):7892-7895. doi: 10.1039/c8cc02760a.

Authors

Keita Omura¹, Yuichiro Aiba, Hiroki Onoda, Joshua Kyle Stanfield, Shinya Ariyasu, Hiroshi Sugimoto, Yoshitsugu Shiro, Osami Shoji, Yoshihito Watanabe

Affiliation

¹ Department of Chemistry, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-0802, Japan. shoji.osami@a.mbox.nagoya-u.ac.jp.

PMID: 29845154
DOI: 10.1039/c8cc02760a

Abstract

Haem substitution is an effective approach to tweak the function of haemoproteins. Herein, we report a facile haem substitution method for self-sufficient cytochrome P450BM3 (CYP102A1) from Bacillus megaterium utilising the transpeptidase Sortase A from Staphylococcus aureus. We successfully constructed Mn-substituted BM3 and investigated its catalytic activity.

MeSH terms

Aminoacyltransferases / chemistry
Aminoacyltransferases / metabolism*
Bacillus megaterium / metabolism
Bacterial Proteins / chemistry
Bacterial Proteins / metabolism*
Base Sequence
Catalysis
Coordination Complexes / chemistry*
Cysteine Endopeptidases / chemistry
Cysteine Endopeptidases / metabolism*
Cytochrome P-450 Enzyme System / chemistry*
Cytochrome P-450 Enzyme System / metabolism
Heme / chemistry
Mutagenesis, Site-Directed
Propane / chemistry
Protein Structure, Tertiary
Sequence Alignment
Staphylococcus aureus / enzymology

Substances

Bacterial Proteins
Coordination Complexes
Heme
Cytochrome P-450 Enzyme System
Aminoacyltransferases
sortase A
Cysteine Endopeptidases
Propane