Project description:Methane is a well-known factor in green house effect and also could be an alternative energy, which produced via methanogenesis by methanogenic archaea. Methanohalophilus portucalensis FDF1T is a halophilic methylotrophic methanogen and has been used as a model to study osmoadaptation in fluctuated abiotic environmental conditions. Here we report a genome-wide phosphoproteomic analysis of M. portucalensis FDF1T to uncover the possible phosphorylation-mediated regulations in methanogenesis and osmoadaptation. According to the biological function analysis revealed 20.6% of total 149 phosphoproteins were involved in the regulation of the single carbon metabolism for methanogenesis and osmoadaptation. Together with genome sequencing and phosphoproteomic analysis, we provided the first evidence to indicate protein phosphorylation networks in regulating the pyrrolysine (Pyl) mediated translation of methylotrophic methyltransferase. Furthermore, the functional assays of glycine sarcosine N-methyltransferase revealed that the enzymatic activity was modulated by threonine phosphorylation. Collectively, our results not only provide new insight into phosphorylation-mediated regulations in single carbon energy metabolism but also clearly demonstrate that phosphorylation plays a critical role in osmolyte betaine synthesis through a phospho-switch mechanism.
Project description:A total gene expression approach was applied to study the methylotrophic nature of B. methanolicus by comparing the gene expression in bacteria grown methylotropic compared to non-methylotrophic. Genes of interest with different gene expression were quantified in the same RNA samples by real-time PCR, confirming the results found in the microarray experiment. Genes of special interest that are expressed higher when grown methylotrophic, were the RuMP pathway genes located on the pBM19.
Project description:A total gene expression approach was applied to study the methylotrophic nature of B. methanolicus by comparing the gene expression in bacteria grown methylotropic compared to non-methylotrophic. Genes of interest with different gene expression were quantified in the same RNA samples by real-time PCR, confirming the results found in the microarray experiment. Genes of special interest that are expressed higher when grown methylotrophic, were the RuMP pathway genes located on the pBM19. Bacillus methanolicus was grown in minimal media with either methanol or mannitol as carbon source. The experiment was preformed in triplicate, with bacterial cultures grown on 3 different days.
