Project description:The degradation of aromatic compounds comprises an important step in the removal of pollutants and re-utilization of plastics and other non-biological polymers. Here we set out to study Pseudomonas sp. strain phDV1, a gram-negative bacterium that was selected for its ability to degrade aromatic compounds. In order to understand how the aromatic compounds and their degradation products are reintroduced in the metabolism of the bacteria and the systematic/metabolic response of the bacterium to the new carbon source, the proteome of this strain was analysed in the presence of succinate, phenol and o-, m-, p-cresol as sole carbon source. We then applied label-free quantitative proteomics to monitor overall proteome remodeling during metabolic adaptation to different carbon sources. As a reference proteome, we grew the bacteria in succinate and then compared the respective proteomes of bacteria grown on phenol and different cresols. In total, we identified 2295 proteins; 1908 proteins were used for quantification between different growth conditions. We found that 70, 100, 150 and 155 proteins were significantly differentially expressed in cells were grown in phenol, o-, m- and p-cresol-containing medium, respectively. The carbon source affected the synthesis of enzymes related to aromatic compound degradation, and in particular, the enzyme involved in the meta-pathway of monocyclic aromatic compounds degradation. In addition, proteins involved in the production of polyhydroxyalkanoate (PHA), an attractive biomaterial, showed higher expression levels in the presence of monocyclic aromatic compounds.Our results provide for the first time comprehensive information on the proteome response of this strain to monocyclic aromatic compounds.
Project description:Degradation of polycyclic aromatic hydrocarbons (PAHs) such as naphthalene by anaerobic microorganisms is poorly understood. Strain NaphS2, an anaerobic sulfate reducing marine delta-proteobacterium is capable of using naphthalene and the aromatic compound benzoate, as well as pyruvate, as an electron donors in the presence of sulfate. In order to identify genes involved in the naphthalene degradation pathway, we compared gene expression in NaphS2 during growth on benzoate vs. pyruvate, naphthalene vs. pyruvate, and naphthalene vs benzoate.
Project description:Brown rot fungi play an essential role in carbon cycling by decomposing lignocellulose into substrates usable by themselves and other microbes. Interactions between bacteria and fungi can be competitive or beneficial, but these relationships are not well understood because of a lack of good model systems. To model cross-feeding between fungi and bacteria, wood decayed by the brown rot fungus Rhodonia placenta was used as a carbon source for the phototrophic bacterium Rhodopseudomonas palustris. We found that fungal decay products generated by Rda. placenta could be used by R. palustris for growth, and later decay stages contained more usable substrates than early stages. Mass spectrometry identified a range of aromatic and non-aromatic acids, but after 95 days of bacterial growth, R. palustris only consumed non-aromatic acids over the aromatic lignin monomers. Genes involved with aromatic compound degradation were unimportant for growth and RNA sequencing revealed that aromatic compound degradation genes were repressed on decayed wood extract. Randomly barcoded transposon sequencing failed to identify a solitary catabolic pathway used by R. palustris, suggestive of substrate co-utilization, and surprisingly showed that genes involved with copper toxicity were essential. Finally, we found genes involved with biosynthesis of certain cofactors were non-essential on decayed wood extract, suggesting these nutrients were scavenged, and implicating these resources in supporting microbial communities in the environment. This study helps lay the foundation for a model fungal-bacterial system to study interactions that govern these relationships which will be valuable for understanding ecological interactions and industrially relevant co-culture systems.
