Photomanipulation of antibiotic susceptibility and biofilm formation of Escherichia coli heterologously expressing photoactivated adenylyl cyclase

Hiro Yasukawa; Noriko Konno; Yukari Haneda; Baku Yamamori; Mineo Iseki; Mami Shibusawa; Yasushi Ono; Ken-ichi Kodaira; Hisashi Funada; Masakatsu Watanabe

doi:10.2323/jgam.58.183

Full Papers

Photomanipulation of antibiotic susceptibility and biofilm formation of Escherichia coli heterologously expressing photoactivated adenylyl cyclase

Hiro Yasukawa, Noriko Konno, Yukari Haneda, Baku Yamamori, Mineo Iseki, Mami Shibusawa, Yasushi Ono, Ken-ichi Kodaira, Hisashi Funada, Masakatsu Watanabe

Author information

Hiro Yasukawa
Graduate School of Science and Engineering, University of Toyama
Noriko Konno
Graduate School of Science and Engineering, University of Toyama
Yukari Haneda
Graduate School of Science and Engineering, University of Toyama
Baku Yamamori
Graduate School of Science and Engineering, University of Toyama
Mineo Iseki
Department of Pharmaceutical Sciences, Toho University
Mami Shibusawa
Graduate School of Bioagricultural Sciences, Nagoya University
Yasushi Ono
Research Center for Basic Research and Development in Natural Sciences, University of Toyama
Ken-ichi Kodaira
Graduate School of Science and Engineering, University of Toyama
Hisashi Funada
Graduate School of Medicine and Pharmaceutical Science, University of Toyama
Masakatsu Watanabe
Graduate School for the Creation of New Photonics Industries

Keywords: antibiotics, biofilm, cAMP, Escherichia coli, photomanipulation

JOURNAL FREE ACCESS

2012 Volume 58 Issue 3 Pages 183-190

DOI https://doi.org/10.2323/jgam.58.183

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Abstract

A cyaA-deficient Escherichia coli strain was transformed by a plasmid carrying the gene for BsPAC, a photoactivated adenylyl cyclase identified from a Beggiatoa sp., and was subjected to an antibiotic susceptibility assay and biofilm formation assay under a light or dark condition. Cells expressing BsPAC that were incubated under blue light (470 nm) were more susceptible to fosfomycin, nalidixic acid and streptomycin than were cells incubated in the dark. Cells expressing BsPAC formed more biofilms when incubated under the light than did cells cultured in the dark. We concluded from these observations that it is possible to determine the importance of cAMP in antibiotic susceptibility and biofilm formation of E. coli by photomanipulating the cellular cAMP level by the use of BsPAC. A site-directed mutant of BsPAC in which Tyr⁷ was replaced by Phe functioned even in the dark, indicating that Tyr⁷ plays an important role in photoactivation of BsPAC. Results of mutational analysis of BsPAC should contribute to an understanding of the molecular basis for photoactivation of the protein.

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