Choose Your Own Adventure: A Comprehensive Database of Reactions Catalyzed by Cytochrome P450 BM3 Variants

Abstract

Cytochrome P450 BM3 monooxygenase is the topic of extensive research as many researchers have evolved this enzyme to generate a variety of products. However, the abundance of information on increasingly diversified variants of P450 BM3 that catalyze a broad array of chemistry is not in a format that enables easy extraction and interpretation. We present a database that categorizes variants by their catalyzed reactions and includes details about substrates to provide reaction context. This database of >1500 P450 BM3 variants is downloadable and machine-readable and includes instructions to maximize ease of gathering information. The database allows rapid identification of commonly reported substitutions, aiding researchers who are unfamiliar with the enzyme in identifying starting points for enzyme engineering. For those actively engaged in engineering P450 BM3, the database, along with this review, provides a powerful and user-friendly platform to understand, predict, and identify the attributes of P450 BM3 variants, encouraging the further engineering of this enzyme.

Design of a genetically programmable and customizable protein scaffolding system for the hierarchical assembly of robust, functional macroscale materials

“…As a proof-of-concept, we selected as our first enzyme for scaffold attachment the challenging, yet extensively studied and highly versatile multi-domain cytochrome P450 monooxygenase CYP102A1 known as P450BM3 18,33 . Unlike many P450 enzymes, P450BM3 can be recombinantly expressed as soluble protein, is self-sufficient due to a fused NADPH reductase domain, is known to catalyzes diverse reactions and over thousand variants have been described 18,33 . Yet, industrial use of this enzyme remains challenging due to its complex, two-domain and dimeric protein structure, undesired “uncoupling” of NADPH derived electrons to produce reactive and inactivating oxygen species and/or H 2 O 2 .…”

“…As a proof-of-concept, we selected as our first enzyme for scaffold attachment the challenging, yet extensively studied and highly versatile multi-domain cytochrome P450 monooxygenase CYP102A1 known as P450BM3 18,33 . Unlike many P450 enzymes, P450BM3 can be recombinantly expressed as soluble protein, is self-sufficient due to a fused NADPH reductase domain, is known to catalyzes diverse reactions and over thousand variants have been described 18,33 .…”