Regulatory transcription factor (CooA)-driven carbon monoxide partial pressure sensing whole-cell biosensor

Byeongchan Kang; Hyeryeong Lee; Soyoung Oh; Ji-Yeon Kim; Young-Joon Ko; In Seop Chang

doi:10.1016/j.heliyon.2023.e17391

Regulatory transcription factor (CooA)-driven carbon monoxide partial pressure sensing whole-cell biosensor

Heliyon. 2023 Jun 21;9(6):e17391. doi: 10.1016/j.heliyon.2023.e17391. eCollection 2023 Jun.

Authors

Byeongchan Kang¹, Hyeryeong Lee^{1

2}, Soyoung Oh¹, Ji-Yeon Kim^{1

2}, Young-Joon Ko¹, In Seop Chang^{1

2}

Affiliations

¹ School of Earth Sciences and Environmental Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Republic of Korea.
² Research Center for Innovative Energy and Carbon Optimized Synthesis for Chemicals (inn-ECOSysChem), Gwangju Institute of Science and Technology, Gwangju 61005, Republic of Korea.

Abstract

We designed and constructed a whole-cell biosensor capable of detecting the presence and quantity of carbon monoxide (CO) using the CO regulatory transcription factor. This biosensor utilizes CooA, a CO-sensing transcription regulator that activates the expression of carbon monoxide dehydrogenase (CODH), to detect the presence of CO and respond by triggering the expression of a GUS reporter protein (β-glucuronidase). The GUS reporter protein is expressed from a CO-induced CooA-binding promoter (P_cooF) by CooA and enables the effective colorimetric detection of CO. An Escherichia coli strain used to validate the biosensor showed growth and GUS activity under anaerobic conditions; this study used the inert gas (Ar) to create anaerobic conditions. The pBRCO biosensor could successfully detect the presence of CO in the headspace. Moreover, the GUS-specific activity of pBRCO according to the CO strength as partial pressure followed Michaelis-Menten kinetics (R² = 0.98). It was confirmed that the GUS-specific activity of pBRCO increased linearly up to 30.39 kPa (R² = 0.98), and thus, a quantitative analysis of CO concentration (i.e., partial pressure) was possible.

Keywords: Carbon monoxide; CooA; GUS reporter gene; Transcription factor; Whole-cell biosensor.