Mediated electron transfer in a photo-bioreactor: continuous flow hydroxylation using cytochrome P450 BM3 in NADPH-free conditions

Fendri, Ali; Valikhani, Donya; Pelletier, Joelle N.

doi:10.1039/d3re00569k

Cited by 3 publications

(1 citation statement)

References 89 publications

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“… 80 An appealing feature of P450 BM3, as a biocatalyst, is that it is not membrane-bound but is expressed in a soluble form. As a result, it can be functionally expressed in large quantities in Escherichia coli and can be stabilized by immobilization, 81 making it suitable for large-scale bioproduction applications. 82 − 85 This further highlights the potential of P450 BM3 as a valuable tool for synthesizing a diverse range of compounds through sustainable and eco-friendly means.…”

Section: P450 Bm3: Properties and Limitationsmentioning

confidence: 99%

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

Fansher,

Besna,

Fendri

et al. 2024

ACS Catal.

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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.

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Section: P450 Bm3: Properties and Limitationsmentioning

confidence: 99%

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

Fansher,

Besna,

Fendri

et al. 2024

ACS Catal.

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Engineering Regioselectivity of P450 BM3 Enables the Biosynthesis of Murideoxycholic Acid by 6β‐Hydroxylation of Lithocholic Acid

Deng,

Zhou,

et al. 2024

Biotechnology Journal

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Murideoxycholic acid (MDCA), as a significant secondary bile acid derived from the metabolism of α/β‐muricholic acid in rodents, is an important component in maintaining the bile acid homeostasis. However, the biosynthesis of MDCA remains a challenging task. Here, we present the development of cytochrome P450 monooxygenase CYP102A1 (P450 BM3) from Bacillus megaterium, employing semi‐rational protein engineering technique. Following three rounds of mutagenesis, a triple variant (T260G/G328A/L82V) has been discovered that proficiently catalyzes the 6β‐hydroxylation of lithocholic acid (LCA), thereby generating MDCA with an impressive 8.5‐fold increase in yield compared to the template P450 BM3 mutant. The MDCA selectivity has been also promoted from 62.0% to 96.3%. This biocatalyst introduces a novel approach for the biosynthesis of MDCA from LCA. Furthermore, molecular docking and dynamics simulations have been employed to unravel the molecular mechanisms underlying the enhanced LCA conversion and MDCA selectivity.

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Indigo production identifies hotspots in cytochrome P450 BM3 for diversifying aromatic hydroxylation

Fansher,

Besna,

Pelletier

2024

Faraday Discuss.

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Mediated electron transfer in a photo-bioreactor: continuous flow hydroxylation using cytochrome P450 BM3 in NADPH-free conditions

Abstract: We designed a proof-of-concept photo-bioreactor enabling continuous flow NADPH-free activation of cytochrome P450 BM3 via mediated electron transfer.

Cited by 3 publications

References 89 publications

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

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

Engineering Regioselectivity of P450 BM3 Enables the Biosynthesis of Murideoxycholic Acid by 6β‐Hydroxylation of Lithocholic Acid

Indigo production identifies hotspots in cytochrome P450 BM3 for diversifying aromatic hydroxylation

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