NICOTINE-INDUCED PROTEOME OF SURPRINSINGLY USEFULL MICROORGANISM: PAENARTHAROBACTER NICOTINOVORANS pAO1.

6-hidroxy-L-nicotine is a metabolic intermediate found in the nicotine catabolic pathway encoded by the pAO1 megaplasmid of Paenarthrobacter nicotinovorans. Extensive work has shown that 6-hidroxy-L-nicotine has shown a great potential as a neuroprotective drug. Thereby, a biotechnology based on Paenarthrobacter nicotinovorans is currently under development for the conversion of nicotine containing waste 6-hidroxy-L-nicotine and other green chemicals. Using P. nicotinovorans for biotechnological applications is nevertheless hindered by the lack of information on how the cells cope with the accumulation and toxicity of the resulting nicotine metabolic by-products. In order to address this issue at the protein level, we performed a proteomics study using reversed phase nanoliquid chromatography tandem mass spectrometry (nanoLC-MS/MS) was performed. P. nicotinovorans was grown on different carbon sources, including nicotine and the cells were harvested at 3 different time intervals: 7, 10 and 24 hours post inoculation. The cells were lyzed, cell free extracts were prepared the proteins were identified using a gel-based proteomics approach employing a NanoAcquity UPLC (Waters, Milford, MA, USA) coupled to a Q-TOF Xevo G2 MS (Waters). Data analysis was performed using Mascot v.2.5.1 (Matrix Science, London, UK) and Scaffold (v.4.8.2, Proteome Software Inc., Portland, OR, USA). This approach allowed us to produce two different sets of proteomics data: PXD008751 (ProteomeXchange Consortium) describing the bacterial proteome on different carbon sources and containing 792 non-redundant proteins as well as PXD012577 (PRIDE) describing a time-based evolution of the nicotine-related proteome that contains 584 non-redundant proteins with an FDR of 0.3%. The differences in protein abundance in the different growth conditions showed that deamination is preferred in the nicotine pathway when citrate can be used as a carbon source. Several putative genes from the pAO1 megaplasmid have been shown to have a nicotine-dependent expression, including a hypothetical polyketide cyclase. This data provides insights into bacterial cells adaptation to the nicotine metabolic intermediates that are known to be toxic and to accumulate in the growth medium. [ABSTRACT FROM AUTHOR]