Sequence modifications and heterologous expression of eukaryotic cytochromes P450 in Escherichia coli

Journal of Bioscience and Bioengineering
Hirofumi IchinoseYukiyo Yamauchi

Abstract

The development of a heterologous expression system is often a principal step in biochemical and biotechnological studies on cytochromes P450 (P450s). However, heterologous expression of eukaryotic membrane-bound P450s in Escherichia coli is still a trial-and-error process because theoretical and systematical experimental procedures have not yet been established. In this study, we generated a series of chimeric variants of fungal P450s by replacing their N-terminal domains with the N-terminal domains of other P450s and explored their potentials for heterologous expression in E. coli. Large-scale screening demonstrated that a number of fungal P450s could be expressed when their N-terminal amino acid sequences were replaced with the corresponding domain of CYP5144C1, even when the expression of the non-chimeric sequence was unpromising. Furthermore, a comprehensive screening resulted in the identification of 64 different types of chimeric partners whose N-terminal domains could potentially be used to increase the expression levels of various P450s. These findings will help to elaborate experimental strategies for high-level heterologous expression of a variety of eukaryotic membrane-bound P450s in E. coli.

References

Jul 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·H J BarnesM R Waterman
May 29, 2002·Phytochemistry·Richard A Dixon, Daneel Ferreira
Jul 18, 2002·Nature Reviews. Drug Discovery·F Peter Guengerich
Dec 19, 2003·Protein Expression and Purification·Dongmei ChengWayne L Backes
Mar 18, 2004·The Journal of Biological Chemistry·Tomoaki YamasakiYoshihiko Ohyama
Mar 7, 2006·Journal of Biotechnology·Rita Bernhardt
May 27, 2006·Current Drug Metabolism·Chul-Ho YunTaeho Ahn
Jun 9, 2006·Trends in Biotechnology·Vlada B Urlacher, Sabine Eiben
Apr 18, 2007·Nature Chemical Biology·Michelle C Y ChangJay D Keasling
Jun 19, 2009·Xenobiotica; the Fate of Foreign Compounds in Biological Systems·A ShuklaE M J Gillam
Apr 2, 2010·Archives of Microbiology·K H M Nazmul Hussain NazirHiroyuki Wariishi
Apr 14, 2010·The Journal of Biological Chemistry·Christal D Sohl, F Peter Guengerich
Dec 31, 2011·Archives of Biochemistry and Biophysics·Hirofumi Ichinose, Hiroyuki Wariishi
Jan 9, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·David R Nelson
Jan 9, 2013·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Steven L Kelly, Diane E Kelly
May 11, 2013·Protein Science : a Publication of the Protein Society·Daniel R McDougleAditi Das
Jul 28, 2013·Biochemical and Biophysical Research Communications·Hirofumi Ichinose, Hiroyuki Wariishi

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Citations

Jul 20, 2016·Microbial Cell Factories·Pradeepraj DurairajHyungdon Yun
Feb 14, 2018·Bioscience Reports·Johanna HausjellOliver Spadiut
Mar 5, 2021·Microbial Biotechnology·Carmen Felpeto-SanteroJosé Luis García
Mar 13, 2021·Natural Product Reports·Xingwang ZhangShengying Li
Aug 8, 2021·Microorganisms·Carmen Felpeto-SanteroJosé Luis García

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