Abstract
The present study examined the aerobic metabolism of trimethylamine in Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine. In both conditions, the trimethylamine was used as a nitrogen source and also accumulated in the cell, slowing the bacterial growth. Decreased bacterial growth was counteracted by the addition of AlCl3. Cell-free extracts prepared from cells grown aerobically on tetradecyltrimethylammonium bromide exhibited trimethylamine monooxygenase activity that produced trimethylamine N-oxide and trimethylamine N-oxide demethylase activity that produced dimethylamine. Cell-free extracts from cells grown on trimethylamine exhibited trimethylamine dehydrogenase activity that produced dimethylamine, which was oxidized to methanal and methylamine by dimethylamine dehydrogenase. These results show that this bacterial strain uses two enzymes to initiate the oxidation of trimethylamine in aerobic conditions. The apparent Km for trimethylamine was 0.7 mM for trimethylamine monooxygenase and 4.0 mM for trimethylamine dehydrogenase, but both enzymes maintain similar catalytic efficiency (0.5 and 0.4, respectively). Trimethylamine dehydrogenase was inhibited by trimethylamine from 1 mM. Therefore, the accumulation of trimethylamine inside Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine may be due to the low catalytic efficiency of trimethylamine monooxygenase and trimethylamine dehydrogenase.
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GIL is Career Members of the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). ASL is grateful for fellowships from CONICET-MCyT Córdoba. This work was supported by grants from ANPCyT-UNRC, MCyT Córdoba and SECYT–UNRC of Argentina.
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Communicated by Stuart Ferguson.
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Liffourrena, A.S., Salvano, M.A. & Lucchesi, G.I. Pseudomonas putida A ATCC 12633 oxidizes trimethylamine aerobically via two different pathways. Arch Microbiol 192, 471–476 (2010). https://doi.org/10.1007/s00203-010-0577-5
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DOI: https://doi.org/10.1007/s00203-010-0577-5