Home A tailored cytochrome P450 monooxygenase from Gordonia rubripertincta CWB2 for selective aliphatic monooxygenation
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A tailored cytochrome P450 monooxygenase from Gordonia rubripertincta CWB2 for selective aliphatic monooxygenation

  • Fabian Peter Josef Schultes ORCID logo , Leon Welter , Myra Schmidtke , Dirk Tischler ORCID logo and Carolin Mügge ORCID logo EMAIL logo
Published/Copyright: September 30, 2024
Biological Chemistry
From the journal Biological Chemistry Volume 405 Issue 9-10

Abstract

Cytochrome P450 monooxygenases are recognized as versatile biocatalysts due to their broad reaction capabilities. One important reaction is the hydroxylation of non-activated C–H bonds. The subfamily CYP153A is known for terminal hydroxylation reactions, giving access to functionalized aliphatics. Whilst fatty derivatives may be converted by numerous enzyme classes, midchain aliphatics are seldomly accepted, a prime property of CYP153As. We report here on a new CYP153A member from the genome of the mesophilic actinobacterium Gordonia rubripertincta CWB2 as an efficient biocatalyst. The gene was overexpressed in Escherichia coli and fused with a surrogate electron transport system from Acinetobacter sp. OC4. This chimeric self-sufficient whole-cell system could perform hydroxylation and epoxidation reactions: conversions of C6–C14 alkanes, alkenes, alcohols and of cyclic compounds were observed, yielding production rates of, e.g., 2.69 mM h−1 for 1-hexanol and 4.97 mM h−1 for 1,2-epoxyhexane. Optimizing the linker compositions between the protein units led to significantly altered activity. Balancing linker length and flexibility with glycine-rich and helix-forming linker units increased 1-hexanol production activity to 350 % compared to the initial linker setup with entirely helical linkers. The study shows that strategic coupling of efficient electron supply and a selective enzyme enables previously challenging monooxygenation reactions of midchain aliphatics.

Keywords: oxyfunctionalization; self-sufficient P450 monooxygenase; linker fusion; actinobacteria; biotransformation
Corresponding author: Carolin Mügge, Microbial Biotechnology, Faculty of Biology and Biotechnology, Ruhr University Bochum, D-44801 Bochum, Germany, E-mail:

Funding source: Bundesministerium für Bildung und Forschung

Award Identifier / Grant number: 03INT513B

Acknowledgments

The experimental help for carbon monoxide assays by Astrit Veluji (Ruhr University Bochum) is greatly appreciated.

  1. Research ethics: Nothing to declare.

  2. Author contributions: Fabian Peter Josef Schultes (formal analysis: lead; investigation: equal; data curation: equal; supervision: supporting; visualization: equal; writing – original draft: lead; writing – review & editing: supporting). Leon Welter (formal analysis: supporting; investigation: equal; writing – original draft: supporting). Myra Schmidtke (formal analysis: supporting; investigation: supporting). Dirk Tischler (funding acquisition: supporting; methodology: supporting; resources: equal; supervision: equal; writing – review & editing: supporting). Carolin Muegge (conceptualization: lead; data curation: equal; visualization: equal; funding acquisition: lead; methodology: lead; project administration: lead; resources: equal; supervision: lead; writing – original draft: supporting; writing – review & editing: lead).

  3. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  4. Competing interests: The authors state no conflict of interest.

  5. Research funding: The authors gratefully acknowledge funding by the German Federal Ministry of Education and Research (BMBF, Grant No. 03INT513B).

  6. Data availability: The raw data can be obtained on request from the corresponding author.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/hsz-2024-0041).

Received: 2024-03-07
Accepted: 2024-09-04
Published Online: 2024-09-30
Published in Print: 2024-10-28

© 2024 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Highlight: GBM Young Investigators Part 5
  3. Highlight: young research groups in Germany – 5th edition
  4. Protein persulfidation in plants: mechanisms and functions beyond a simple stress response
  5. Pathological and physiological roles of ADP-ribosylation: established functions and new insights
  6. Implications of TRPM3 and TRPM8 for sensory neuron sensitisation
  7. The complex regulation of Slo1 potassium channels from a structural perspective
  8. The TOM complex from an evolutionary perspective and the functions of TOMM70
  9. Insights in caveolae protein structure arrangements and their local lipid environment
  10. Insights into caudate amphibian skin secretions with a focus on the chemistry and bioactivity of derived peptides
  11. Analysis of cell cycle stage, replicated DNA, and chromatin-associated proteins using high-throughput flow cytometry
  12. A tailored cytochrome P450 monooxygenase from Gordonia rubripertincta CWB2 for selective aliphatic monooxygenation
https://doi.org/10.1515/hsz-2024-0041
Keywords for this article
oxyfunctionalization; self-sufficient P450 monooxygenase; linker fusion; actinobacteria; biotransformation

Articles in the same Issue

  1. Frontmatter
  2. Highlight: GBM Young Investigators Part 5
  3. Highlight: young research groups in Germany – 5th edition
  4. Protein persulfidation in plants: mechanisms and functions beyond a simple stress response
  5. Pathological and physiological roles of ADP-ribosylation: established functions and new insights
  6. Implications of TRPM3 and TRPM8 for sensory neuron sensitisation
  7. The complex regulation of Slo1 potassium channels from a structural perspective
  8. The TOM complex from an evolutionary perspective and the functions of TOMM70
  9. Insights in caveolae protein structure arrangements and their local lipid environment
  10. Insights into caudate amphibian skin secretions with a focus on the chemistry and bioactivity of derived peptides
  11. Analysis of cell cycle stage, replicated DNA, and chromatin-associated proteins using high-throughput flow cytometry
  12. A tailored cytochrome P450 monooxygenase from Gordonia rubripertincta CWB2 for selective aliphatic monooxygenation
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