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Exploring the Biocatalytic Potential of a Self-Sufficient Cytochr ome P450 from Thermothelomyces thermophila

Fürst, M., Kerschbaumer, B., Rinnofner, C., Migglautsch, A., Winkler, M. & Fraaije, M., 6-Jun-2019, In : Advanced Synthesis & Catalysis. 361, 11, p. 2487–2496 10 p.

Research output: Contribution to journalArticleAcademicpeer-review

Among nature's arsenal of oxidative enzymes, cytochrome P450s (CYPs) catalyze the most challenging reactions, the hydroxylations of non‐activated C−H bonds. Human CYPs are studied in drug development due to their physiological role at the forefront of metabolic detoxification, but their challenging handling makes them unsuitable for application. CYPs have a great potential for biocatalysis, but often lack appropriate features such as high and soluble expression, self‐sufficient internal electron transport, high stability, and an engineerable substrate scope. We have probed these characteristics for a recently described CYP that originates from the thermophilic fungus Thermothelomyces thermophila (CYP505A30), a homolog of the well‐known P450‐BM3 from Bacillus megaterium. CYP505A30 is a natural monooxygenase‐reductase fusion, is well expressed, and moderately tolerant towards temperature and solvent exposure. Although overall comparable, we found the stability of the enzyme's domains to be inverse to P450‐BM3, with a more stable reductase compared to the heme domain. After analysis of a homology model, we created mutants of the enzyme based on literature data for P450‐BM3. We then probed the enzyme variants in bioconversions using a panel of active pharmaceutical ingredients, and activities were detected for a number of structurally diverse compounds. Ibuprofen was biooxidized in a preparative scale whole cell bioconversion to 1‐, 2‐ and 3‐hydroxyibuprofen.
Original languageEnglish
Pages (from-to)2487–2496
Number of pages10
JournalAdvanced Synthesis & Catalysis
Volume361
Issue number11
Early online date19-Mar-2019
Publication statusPublished - 6-Jun-2019

    Keywords

  • HYDROXYLATION, IBUPROFEN, SYSTEM, MONOOXYGENASE, EPOXIDATION, CAPSAICIN, OXIDATION, P450BM3, ACID

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