Arginine deiminase pathway provides ATP and boosts growth of the gas-fermenting acetogen Clostridium autoethanogenum Metabolic Engineering, http://dx.doi.org/10.1016Engineering, http://dx.doi.org/10. /j.ymben.2017 This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting galley proof before it is published in its final citable form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. AbstractAcetogens are attractive organisms for the production of chemicals and fuels from inexpensive and non-food feedstocks such as syngas (CO, CO 2 and H 2 ). Expanding their product spectrum beyond native compounds is dictated by energetics, particularly ATP availability. Acetogens have evolved sophisticated strategies to conserve energy from reduction potential differences between major redox couples, however, this coupling is sensitive to small changes in thermodynamic equilibria. To accelerate the development of strains for energy-intensive products from gases, we used a genome-scale metabolic model (GEM) to explore alternative ATP-generating pathways in the gas-fermenting acetogen 1 Present address: Department of Plant and Environmental Science, University of Copenhagen, Copenhagen, Denmark Clostridium autoethanogenum. Shadow price analysis revealed a preference of C.autoethanogenum for nine amino acids. This prediction was experimentally confirmed under heterotrophic conditions. Subsequent in silico simulations identified arginine (ARG) as a key enhancer for growth. Predictions were experimentally validated, and faster growth was measured in media containing ARG (t D~4 h) compared to growth on yeast extract (t D~9 h). The growth-boosting effect of ARG was confirmed during autotrophic growth.Metabolic modelling and experiments showed that acetate production is nearly abolished and fast growth is realised by a three-fold increase in ATP production through the arginine deiminase (ADI) pathway. The involvement of the ADI pathway was confirmed by metabolomics and RNA-sequencing which revealed a ~500-fold up-regulation of the ADI pathway with an unexpected down-regulation of the Wood-Ljungdahl pathway. The data presented here offer a potential route for supplying cells with ATP, while demonstrating the usefulness of metabolic modelling for the discovery of native pathways for stimulating growth or enhancing energy availability.
Computer simulations were performed to study the characteristic transition temperatures of the adsorbate monolayer transitions on graphite and to determine the layering temperatures for higher layers at temperatures less than the bulk triple point temperature. Two models for graphite were studied to examine the effects of finite size of the graphene layer on the evolution of the characteristics of the monolayer, its boundary with the gas phase, and the resulting isotherm and isosteric heat versus loading. Both models give good agreement with experiment for the 2D-critical point and the 2D-triple point, but the finite model is more successful in representing the experimental isotherm and isosteric heat. Radial density distribution for the monolayer supports this, and it illustrates the manner in which the monolayer is compressed with loading, by mass transfer of molecules from the gas phase through the 1D-boundary of the 2D monolayer adsorbate. As the adsorbed phase grows beyond the monolayer, the structure of the thick adsorbed film was shown to lie between the crystalline structure and the dense supercooled liquid, as reflected in partial wetting, defined as finite loading of the adsorbed film at the bulk sublimation pressure.
We have carried out an extensive grand canonical Monte Carlo simulation to investigate the adsorption of neon and xenon on graphite. The adsorbate collision diameters of neon and xenon are smaller and greater respectively, than the commensurate graphite lattice spacing λ=3×3R30 of 0.426 nm. Simulated isotherms and isosteric heats were obtained using a graphite model that has been shown to describe successfully the adsorbate transitions for krypton, methane and nitrogen by Prasetyo et al. (2017), which have collision diameters close to λ. Neon does not exhibit commensurate (C) packing because the gain in the intermolecular potential interactions in the incommensurate (IC) packing when molecules move away from carbon hexagon centres, does not compensate for the increase in the solid-fluid potential energy. Xenon, on the other hand, exhibits IC packing because its molecular size is greater than λ. Nevertheless, at a sufficiently high chemical potential, the first layer of xenon changes from the IC to C packing (in contrast to what is observed for krypton, nitrogen and methane). This transition occurs because the decrease in the xenon intermolecular interactions is sufficiently compensated by the increase in the solid-fluid interaction, and the increase in the fluid-fluid interactions between molecules in the first layer and those in the second layer. This finding is supported by the X-ray diffraction study by Mowforth et al. (1986) and Morishige et al. (1990).
Simulations of ammonia adsorption on graphite were carried out over a range of temperatures to investigate the transition from nonwetting to wetting. The process is governed by a subtle interplay between the various interactions in the system and the temperature. At temperatures below the bulk triple point, the system is nonwetting; above the triple point, we observed continuous wetting, preceded by a prewetting region in which the so-called thin-to-thick film transition occurs. This system serves as an excellent example of wetting/nonwetting behavior in an associating fluid as a function of temperature because the heat of sublimation (or condensation) is greater than the isosteric heat of adsorption at zero loading. The nonwetting-to-wetting transition (NW/W) is also strongly affected by the adsorbate–adsorbate interaction, which becomes important when this contribution to the isosteric heat is of a similar magnitude to the heat of condensation. An appropriate indicator of a NW/W transition at a given loading is therefore the difference between the isosteric heat and the heat of sublimation (or condensation). Our simulation results show the “thin-to-thick” film transition in the temperature range between 195 and 240 K, which has not been previously explained. Above 240 K, continuous wetting occurs. This study provides a basis for a better understanding of adsorption in a range of systems because ammonia is an intermediate between simple molecules, such as argon, and strongly associating fluids, such as water.
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