Functional Expression of a Mo-Cu-Dependent Carbon Monoxide Dehydrogenase (CODH) and Its Use as a Dissolved CO Bio-microsensor
| Autor: | Deby Fapyane, Michael Etzerodt, Stacy Simai Reginald, In Seop Chang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
COmicrosensor
Mo-Cu carbon monoxide dehydrogenase (Mo-Cu CODH) Bioengineering Electrolyte Catalysis Metal Comamonadaceae chemistry.chemical_compound Multienzyme Complexes Escherichia coli Instrumentation Fluid Flow and Transfer Processes Detection limit Carbon Monoxide Clark-type sensor biology Process Chemistry and Technology biology.organism_classification CO oxidation Aldehyde Oxidoreductases CO bio-microsensor Solvent Hydrogenophaga pseudoflava chemistry visual_art Ionic liquid visual_art.visual_art_medium biology.protein Carbon monoxide dehydrogenase Nuclear chemistry |
| Zdroj: | Reginald, S S, Etzerodt, M, Fapyane, D & Chang, I S 2021, ' Functional Expression of a Mo-Cu-Dependent Carbon Monoxide Dehydrogenase (CODH) and Its Use as a Dissolved CO Bio-microsensor ', ACS Sensors, vol. 6, no. 7, pp. 2772–2782 . https://doi.org/10.1021/acssensors.1c01243 |
| ISSN: | 2379-3694 |
| Popis: | Herein, we report the heterologous expression in Escherichia coli of a Mo-Cu-containing carbon monoxide dehydrogenase (Mo-Cu CODH) from Hydrogenophaga pseudoflava, which resulted in an active protein catalyzing CO oxidation to CO2. By supplying the E. coli growth medium with Na2MoO4 (Mo) and CuSO4 (Cu), the Mo-Cu CODH metal cofactors precursors, the expressed L-subunit was found to have CO-oxidation activity even without the M- and S- subunits. This successful expression of CO-oxidizing-capable single L-subunit provides direct evidence of its role as the catalytic center of Mo-Cu CODH that has not been discovered and studied before. Subsequently, we used the expressed protein to construct a CO bio-microsensor based on a newly developed fast and sensitive Clark-type CO2 transducer using an aprotic solvent/ionic liquid electrolyte. The CO bio-microsensor exhibited a linear response to CO concentration in the 0-9 μM range, with a limit of detection (LOD) of 15 nM CO. The sensor uses a mixture of Mo-Cu CODH's L-subunit/Mo, Cu cofactors/methylene blue, confined in the enzyme chamber that is placed in front of a CO2 transducer. The optimized sensor's sensitivity and performance were retained to levels of at least 80% for 1 week of continuous polarization and operation in an aqueous medium. We have also demonstrated the use of an alkaline front-trap solution to make a completely O2/CO2 interference-free microsensor. The CO bio-microsensor developed in this study is potentially useful as an analytical tool for the detection of trace CO in dissolved form for monitoring dissolved CO concentration dynamics in natural or synthetic systems. |
| Databáze: | OpenAIRE |
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