Project description:ra15-05_tt1 - tt1-gr - Discover TT1 donwstream targets - An inducible line carrying TT1, an early marker of endothelium development, fused to the rat glucocorticoid receptor (GR), that allows a dexamethasone (DEX)-mediated posttranslational induction of the proteins, has been created. Siliques have been first treated with cycloheximide, an inhibitor of protein synthesis, to prevent indirect transcriptional effects of the inducible TT1 transcription factor. RNA profiling experiments (CATMA microarrays) will be then conducted before and after DEX induction of TT1. These experiments will unveil direct downstream target genes of the TT1 transcription factor.
Project description:The application of chemical dispersants during marine oil spills can affect the community composition and activity of native marine microorganisms. Several studies have indicated that certain marine hydrocarbon-degrading bacteria, such as Marinobacter spp., can be inhibited by chemical dispersants, resulting in lower abundances and/or reduced hydrocarbon-biodegradation rates. In this respect, a major knowledge gap exists in understanding the mechanisms underlying these observed physiological effects. Here, we performed comparative proteomics of the Deepwater Horizon isolate Marinobacter sp. TT1 grown under different conditions that varied regarding the supplied carbon sources (pyruvate vs. n-hexadecane) and whether or not dispersant (Corexit EC9500A) was added, or that contained crude oil in the form of a water-accommodated fraction (WAF) or chemically-enhanced WAF (CEWAF). We characterized the proteins associated with alkane metabolism and alginate biosynthesis in strain TT1, report on its potential for aromatic hydrocarbon biodegradation and present a proposed metabolism of Corexit components as carbon substrates for the strain. Our findings implicate Corexit in affecting hydrocarbon metabolism, chemotactic motility, biofilm formation, and inducing solvent tolerance mechanisms like efflux pumps in strain TT1. This study provides novel insights into dispersant impacts on microbial hydrocarbon degraders that should be taken into consideration for future oil spill response actions.
Project description:Proteomes of Vibrio sp. 1A01 exponentially growing on various carbon sources as well as on chitin. Chitin samples contain planktonic cells, particle-associated proteins and excreted proteins.
Project description:Brown macroalgae holds an enormous potential as a future feedstock because it rapidly forms large biomasses and has high carbohydrate content (35% of its dry weight consists of alginate and mannitol). However, utilization of brown macroalgae by conventional microbial platforms (e.g., Escherichia coli and Saccharomyces cerevisiae) has been limited due to the inability of these platforms to metabolize alginate. Although recent studies engineered them to utilize alginate, their growth rates and metabolic activities are still too low for industrial applications, likely due to the unoptimized expression of multiple xenogeneic genes. Here, we isolated Vibrio sp. dhg, a novel, fast-growing bacterium that has been naturally evolved for efficient alginate assimilation (growth rate = 0.98 h-1). Especially, both the growth rate and sugar uptake rate of V. sp. dhg are substantially higher than the rates of E. coli for most biomass-derivable sugars. Based on our systematic characterization of its metabolism and gene expression architecture, we were able to develop a genetic toolbox for its engineering. By using this microorganism, we successfully demonstrated its ability to produce a broad spectrum of chemicals from alginate-mannitol mixtures with high productivities (1.1 g ethanol/L/h, 1.3 g 2,3-butanediol and acetoin/L/h, and 0.69 mg lycopene/L/h). Collectively, the V. sp. dhg strain is a powerful platform for the conversion of brown macroalgae sugars whose usage will dramatically accelerate the production of value-added biochemicals in the future.