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Applied and Environmental Microbiology, May 2001, p. 2136-2138, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2136-2138.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Export of Cytochrome P450 105D1 to the Periplasmic Space of Escherichia coli

Mustak A. Kaderbhai, Cynthia C. Ugochukwu, Steven L. Kelly,* and David C. Lamb

Institute of Biological Sciences, University of Wales Aberystwyth, Aberystwyth SY23 3DA, Wales, United Kingdom

Received 11 December 2000/Accepted 30 January 2001

CYP105D1, a cytochrome P450 from Streptomyces griseus, was appended at its amino terminus to the secretory signal of Escherichia coli alkaline phosphatase and placed under the transcriptional control of the native phoA promoter. Heterologous expression in E. coli phosphate-limited medium resulted in abundant synthesis of recombinant CYP105D1 that was translocated across the bacterial inner membrane and processed to yield authentic, heme-incorporated P450 within the periplasmic space. Cell extract and whole-cell activity studies showed that the periplasmically located CYP105D1 competently catalyzed NADH-dependent oxidation of the xenobiotic compounds benzo[a]pyrene and erythromycin, further revealing the presence in the E. coli periplasm of endogenous functional redox partners. This system offers substantial advantages for the application of P450 enzymes to whole-cell biotransformation strategies, where the ability of cells to take up substrates or discard products may be limited.

* Corresponding author. Mailing address: Institute of Biological Sciences, Edward Llywd Building, University of Wales, Aberystwyth SY23 3DA, Wales, United Kingdom. Phone: (0044) 1970 621515. Fax: (0044) 1970 622350. E-mail: Steven.Kelly{at}aber.ac.uk.

Applied and Environmental Microbiology, May 2001, p. 2136-2138, Vol. 67, No. 5
0099-2240/01/$04.00+0   DOI: 10.1128/AEM.67.5.2136-2138.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Akhtar, M. K., Kaderbhai, N. N., Hopper, D. J., Kelly, S. L., Kaderbhai, M. A. (2003). Export of a Heterologous Cytochrome P450 (CYP105D1) in Escherichia coli Is Associated with Periplasmic Accumulation of Uroporphyrin. J. Biol. Chem. 278: 45555-45562 [Abstract] [Full Text]  
  • Kaderbhai, M. A., Hopper, D. J., Akhtar, K. M., Abbas, S. K., Kaderbhai, N. N. (2003). A Cytochrome c from a Lupanine-Transforming Pseudomonas putida Strain Is Expressed in Escherichia coli during Aerobic Cultivation and Efficiently Exported and Assembled in the Periplasm. Appl. Environ. Microbiol. 69: 4727-4731 [Abstract] [Full Text]  


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