Abstract
Nicotine alkaloid plays an important role in smoking addiction and has been found to be implicated in many biological processes. In addition, large quantities of waste with high concentrations of nicotine are generated during the tobacco‐manufacturing process and are simply discarded into the environment. The enzymes encoded by pAO1 megaplasmid of Arthrobacter nicotinovorans might offer a unique way of exploiting nicotine waste for the production of “green” chemicals and for understanding smoking addiction. Although the organization, function, origin and evolution of the pAO1 megaplasmid have been extensively studied, several key points such as regulation of the expression of the nicotine catabolic genes and nicotine transport or metabolic fate of the pyridine ring are still missing.The current work attempts to identify all the nicotine induced proteins from Arthrobacter nicotinovorans pAO1+ by using using nano LC‐MS/MS. Arthrobacter nicotinovorans pAO1+ cells were grown to log phase on citrate medium and on citrate medium supplemented with 0,005% nicotine. The cells were lysed using the method described by Henne et al., J Proteome Res 2009, 8:1704–16. Cell free extracts were separated on 9–16% SDS‐PAGE maxi (20 × 20 cm) gradient gels and Commasie stained. The extracts from the bacteria grown on nicotine‐containing media showed several extra bands în the range of 60 and 30 kDa. One particularity of Arthrobacter nicotinovorans extract is the high abundance of small molecular weight protein of around 6.5 kDa. Full scale proteomic analysis is currently under way.Identification of all the key regulators and enzymes of the nicotine catabolism in Arthrobacter nicotinovorans would allow a better understand smoking addiction, but to also a way to manipulate the nicotine catabolic pathway to treat tobacco waste and convert it into green chemicals.Support or Funding InformationMM is supported by a Fulbright Senior Award from the the Romanian ‐ U.S. Fulbright Commission, 2017.
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