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Sharpless, W. A.

Publications and source records attributed to Sharpless, W. A..

2 recordsLinked to original sources

Towards Environmental Control of Microbiomes

Microbial communities have consequential effects on health and the environment yet remain uncontrollable due to their complex dynamics. Ecological modeling offers a platform to overcome their nonlinear and interconnected nature but traditionally does not account for context-dependence. Here, we extend the generalized Lotka-Volterra (gLV) model to accommodate a varying environment by identifying how environmental changes alter species growth rates and interactions in a manner that predicts full community trajectories across environmental gradients. We identify key environment-varying interactions within a synthetic community derived from the Oryzae sativa rhizosphere, and demonstrate how variations in the environment change fixed point compositions and rates of convergence. With our model, we simulate how precise perturbations of the environment can offer improvements in an optimal control problem of driving a community to a target composition. We show that environmental perturbation can minimize the total species input (direct species perturbation) and greatly expand the set of initial states from which a desired target can be reached despite stochasticity. This work demonstrates that a formal perspective on environmental influence of community dynamics is valuable for not only understanding seasonal changes or anthropogenic manipulations, but is critical for improving control of the microbiome.

systems biology↗

Massively parallel fitness profiling reveals multiple novel enzymes in Pseudomonas putida lysine metabolism

The lysine metabolism of Pseudomonas putida can produce multiple important commodity chemicals and is implicated in rhizosphere colonization. However, despite intensive study, the biochemical and genetic links between lysine metabolism and central metabolism remain unresolved in P. putida. Here, we leverage Random Barcode Transposon Sequencing (RB-TnSeq), a genome-wide assay measuring the fitness of thousands of genes in parallel, to identify multiple novel enzymes in both L- and D-lysine metabolism. We first describe three pathway enzymes that catabolize 2-aminoadipate (2AA) to 2-ketoglutarate (2KG) connecting D-lysine to the TCA cycle. One of these enzymes, PP_5260, contains a DUF1338 domain, a family without a previously described biological function. We demonstrate PP_5260 converts 2-oxoadipate (2OA) to 2-hydroxyglutarate (2HG), a novel biochemical reaction. We expand on recent work showing that the glutarate hydroxylase, CsiD, can co-utilize both 2OA and 2KG as a co-substrate in the hydroxylation of glutarate. Finally we demonstrate that the cellular abundance of D- and L-lysine pathway proteins are highly sensitive to pathway specific substrates. This work demonstrates the utility of RB-TnSeq for discovering novel metabolic pathways in even well-studied bacteria.\n\nImportanceO_LIP. putida is an attractive host for metabolic engineering as its lysine metabolism can be utilized for the production of multiple important commodity chemicals.\nC_LIO_LIWe demonstrate the first biochemical evidence of a bacterial 2OA catabolic pathway to central metabolites.\nC_LIO_LIDUF1338 proteins are widely dispersed across many kingdoms of life. Here we demonstrate the first biochemical evidence of function for a member of this protein family.\nC_LI

microbiology↗