Inducible utilization pathways reflect widespread microbial strategies to uptake and consume sugars from the environment. Despite their broad importance and extensive characterization, little is known how these pathways naturally respond to their inducing sugar in individual cells. Here, we performed single-cell analyses to probe the behavior of representative pathways in the model bacterium Escherichia coli. We observed diverse single-cell behaviors, including uniform responses (D-lactose, D-galactose, N-acetylglucosamine, N-acetylneuraminic acid), “all-or-none” responses (D-xylose, L-rhamnose), and complex combinations thereof (L-arabinose, D-gluconate). Mathematical modeling and probing of genetically modified pathways revealed that the simple framework underlying these pathways—inducible transport and inducible catabolism—could give rise to most of these behaviors. Sugar catabolism was also an important feature, as disruption of catabolism eliminated tunable induction as well as enhanced memory of previous conditions. For instance, disruption of catabolism in pathways that respond to endogenously synthesized sugars led to full pathway induction even in the absence of exogenous sugar. Our findings demonstrate the remarkable flexibility of this simple biological framework, with direct implications for environmental adaptation and the engineering of synthetic utilization pathways as titratable expression systems and for metabolic engineering.
Only limited information is available regarding the manner in which cyclic carbonate and ester solvents coordinate Li + cations in electrolyte solutions for lithium batteries. One approach to gleaning significant insight into these interactions is to examine crystalline solvate structures. To this end, eight new solvate structures are reported with ethylene carbonate, γ-butyrolactone, and γ-valerolactone: (EC) 3 :LiClO 4 , (EC) 2 :LiClO 4 , (EC) 2 :LiBF 4 , (GBL) 4 :LiPF 6 , (GBL) 1 :LiClO 4 , (GVL) 1 :LiClO 4 , (GBL) 1 :LiBF 4 , and (GBL) 1 :LiCF 3 SO 3 . The crystal structure of (EC) 1 :LiCF 3 SO 3 is also re-reported for comparison. These structures enable the factors that govern the manner in which the ions are coordinated and the ion/solvent packingin the solid-stateto be scrutinized in detail.
Solvate
crystal structures serve as useful models for the molecular-level
interactions within the diverse solvates present in liquid electrolytes.
Although acyclic carbonate solvents are widely used for Li-ion battery
electrolytes, only three solvate crystal structures with lithium salts
are known for these and related solvents. The present work, therefore,
reports six lithium salt solvate structures with dimethyl and diethyl
carbonate, (DMC)2:LiPF6, (DMC)1:LiCF3SO3, (DMC)1/4:LiBF4, (DEC)2:LiClO4, (DEC)1:LiClO4, and
(DEC)1:LiCF3SO3 and four with the
structurally related methyl and ethyl acetate, (MA)2:LiClO4, (MA)1:LiBF4, (EA)1:LiClO4, and (EA)1:LiBF4.
Titratable
systems are common tools in metabolic engineering to
tune the levels of enzymes and cellular components as part of pathway
optimization. For nonmodel microorganisms with limited genetic tools,
inducible sugar utilization pathways offer built-in titratable systems.
However, these pathways can exhibit undesirable single-cell behaviors
that hamper the uniform and tunable control of gene expression. Here,
we applied mathematical modeling and single-cell measurements of l-arabinose utilization in Escherichia coli to systematically explore how sugar utilization pathways can be
altered to achieve desirable inducible properties. We found that different
pathway alterations, such as the removal of catabolism, constitutive
expression of high-affinity or low-affinity transporters, or further
deletion of the other transporters, came with trade-offs specific
to each alteration. For instance, sugar catabolism improved the uniformity
and linearity of the response at the cost of requiring higher sugar
concentrations to induce the pathway. Within these alterations, we
also found that a uniform and linear response could be achieved with
a single alteration: constitutively expressing the high-affinity transporter.
Equivalent modifications to the d-xylose utilization pathway
yielded similar responses, demonstrating the applicability of our
observations. Overall, our findings indicate that there is no ideal
set of typical alterations when co-opting natural utilization pathways
for titratable control and suggest design rules for manipulating these
pathways to advance basic genetic studies and the metabolic engineering
of microorganisms for optimized chemical production.
A Raman spectral evaluation of numerous crystalline solvates
with
lithium perchlorate (LiClO4) has been conducted over a
wide temperature range. Two new solvate crystal structures(PMDETA)1:LiClO4 and (THF)1:LiClO4 with N,N,N′,N″,N″-pentamethyldiethylenetriamine
and tetrahydrofuranhave been determined to aid in this study.
With a help of density functional theory (DFT) and molecular dynamics
(MD) simulations, the spectroscopic data have been correlated with
varying modes of ClO4
–···Li+ cation coordination within the solvate structures to create
a characterization tool to facilitate the Raman band assignments for
the determination of ionic association interactions within solid and
liquid electrolytes containing LiClO4. This study demonstrates
that many of the spectroscopic evaluation conclusions reported in
the scientific literature for LiClO4-based electrolytes
are inaccurate.
Fungal bloodstream infections are a significant problem in the United States, with an attributable mortality rate of up to 40%. An early diagnosis to direct appropriate therapy has been shown to be critical to reduce mortality rates. Conventional phenotypic methods for fungal detection take several days, which is often too late to impact outcomes. Herein, we describe a cost-effective multiplex assay platform for the rapid detection and differentiation of major clinically relevant species directly from blood culture. This approach utilizes a novel biotin-labeled polymer-mediated signal amplification process combined with targeting rRNA to exploit phylogenetic differences for sensitive and unambiguous species identification; this assay detects seven pathogenic species (, ,, ,, , and) simultaneously with very high specificity to the species level in less than 80 min with the limits of detection at 1 × 10 to 10 × 10 CFU/ml or as few as 50 CFU per assay. The performance of the described assay was verified with 67 clinical samples (including mixed multiple-species infections as well), with an overall 100% agreement with matrix-assisted laser desorption ionization (MALDI) mass spectrometry-based reference results. By providing a species identity rapidly, the clinician is aided with information that may direct appropriate therapy sooner and more accurately than current approaches, including PCR-based tests.
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