Abstract Engineering living cells for production of chemicals, enzymes and therapeutics can burden cells due to use of limited native co-factor availability and/or expression burdens, totalling a fitness deficit compared to parental cells encoded through long evolutionary trajectories to maximise fitness. Ultimately, this discrepancy puts a selective pressure against fitness-burdened engineered cells under prolonged bioprocesses, and potentially leads to complete eradication of high-performing engineered cells at the population level. Here we present the mutation landscapes of fitness-burdened yeast cells engineered for vanillin-β-glucoside production. Next, we design synthetic control circuits based on transcriptome analysis and biosensors responsive to vanillin-β-glucoside pathway intermediates in order to stabilize vanillin-β-glucoside production over ∼55 generations in sequential passage experiments. Furthermore, using biosensors with two different modes of action we identify control circuits linking vanillin-β-glucoside pathway flux to various essential cellular functions, and demonstrate control circuits robustness and 92% higher vanillin-β-glucoside production, including 5-fold increase in total vanillin-β-glucoside pathway metabolite accumulation, in a fed-batch fermentation compared to vanillin-β-glucoside producing cells without control circuits.
Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract, please click on HTML or PDF.
Most caddisfly larvae build cases of silk and a variety of collected materials. Multiple functions, including protection from predators, resistance to entrainment by high flows, and improved respiration, have been suggested for caddisfly cases. We investigated the functional role of cases built by Dicosmoecus gilvipes, a limnephilid caddisfly. In this species, the 1st- through 4th-instar larvae build cases with plant material and attach Douglas-fir needles as lateral extensions that resemble vanes on an arrow. We tested whether the lateral extensions and entire case deterred predators by manipulating lateral extensions and case presence for larvae exposed to large steelhead trout. No larva with a case (with or without lateral extensions) was consumed during the experiment, whereas all larvae without a case were consumed. We tested whether lateral extensions provided stability against overturning and entrainment by manipulating presence of lateral extensions and subjecting larvae to turbulent flow conditions. Once dislodged, larvae with lateral extensions experienced fewer revolutions and regained their footing faster than those without extensions. Our results suggest lateral extensions provide stability against overturning in fast flow and may improve the ability of larvae to forage efficiently in turbulent flow conditions. Other caddisfly species build lateral extensions on their case, and the extensions may provide similar benefits for these taxa.
Previously proposed sensor network data dissemination schemes require periodic low-rate flooding of data in order to allow recovery from failure. We consider constructing two kinds of multipaths to enable energy efficient recovery from failure of the shortest path between source and sink. Disjoint multipath has been studied in the liteature. w propose a model braided multipath scheme, which results in several partially disjoint multipath schemes. We find that braided multipaths are a viable alternative for energy-efficient recovery from isolated and patterned failures
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Abstract In pharmaceutical discovery, the “magic methyl” effect describes a substantial improvement in the pharmacological properties of a drug candidate with the incorporation of methyl groups. Therefore, to expedite the synthesis of methylated drug analogs, late-stage, undirected methylations of C(sp 3 ) − H bonds in complex molecules would be valuable. However, current methods for site-selective methylations are limited to activated C(sp 3 ) − H bonds. Here we describe a site-selective, undirected methylation of unactivated C(sp 3 ) − H bonds, enabled by photochemically activated peroxides and a nickel(II) complex whose turnover is enhanced by an ancillary ligand. The methodology displays compatibility with a wide range of functional groups and a high selectivity for tertiary C−H bonds, making it suitable for the late-stage methylation of complex organic compounds that contain multiple alkyl C−H bonds, such as terpene natural products, peptides, and active pharmaceutical ingredients. Overall, this method provides a synthetic tool to explore the “magic methyl” effect in drug discovery.