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At least 865 records · Page 48

Data for "Metabolic Engineering Strategies to Produce Medium-Chain Oleochemicals via Acyl-ACP:CoA Transacylase Activity"

Microbial lipid metabolism is an attractive route for producing oleochemicals. The predominant strategy centers on heterologous thioesterases to synthesize desired chain-length fatty acids. To convert acids to oleochemicals (e.g., fatty alcohols, ketones), the narrowed fatty acid pool needs to be reactivated as coenzyme A thioesters at cost of one ATP per reactivation – an expense that could be saved if the acyl-chain was directly transferred from ACP- to CoA-thioester. Here, we demonstrate such an alternative acyl-transferase strategy by heterologous expression of PhaG, an enzyme first identified in Pseudomonads, that transfers 3-hydroxy acyl-chains between acyl-carrier protein and coenzyme A thioester forms for creating polyhydroxyalkanoate monomers. We use it to create a pool of acyl-CoA’s that can be redirected to oleochemical products. Through bioprospecting, mutagenesis, and metabolic engineering, we develop three strains of Escherichia coli capable of producing over 1 g/L of medium-chain free fatty acids, fatty alcohols, and methyl ketones.

Bioproducts↗

Data for Sugar Accumulation Enhancement in Sorghum Stem is Associated with Reduced Reproductive Sink Strength and Increased Phloem Unloading Activity

Sweet sorghum has emerged as a promising source of bioenergy mainly due to its high biomass and high soluble sugar yield in stems. Studies have shown that loss-of-function Dry locus alleles have been selected during sweet sorghum domestication, and decapitation can further boost sugar accumulation in sweet sorghum, indicating that the potential for improving sugar yields is yet to be fully realized. To maximize sugar accumulation, it is essential to gain a better understanding of the mechanism underlying the massive accumulation of soluble sugars in sweet sorghum stems in addition to the Dry locus. We performed a transcriptomic analysis upon decapitation of near-isogenic lines for mutant (d, juicy stems, and green leaf midrib) and functional (D, dry stems and white leaf midrib) alleles at the Dry locus. Our analysis revealed that decapitation suppressed photosynthesis in leaves, but accelerated starch metabolic processes in stems. SbbHLH093 negatively correlates with sugar levels supported by genotypes (DD vs. dd), treatments (control vs. decapitation), and developmental stages post anthesis (3d vs.10d). D locus gene SbNAC074A and other programmed cell death-related genes were down regulated by decapitation, while sugar transporter-encoding gene SbSWEET1A was induced. Both SbSWEET1A and Invertase 5 were detected in phloem companion cells by RNA in situ assay. Loss of the SbbHLH093 homolog, AtbHLH093, in Arabidopsis led to a sugar accumulation increase. This study provides new insights into sugar accumulation enhancement in bioenergy crops, which can be potentially achieved by reducing reproductive sink strength and enhancing phloem unloading.

Transcriptomics↗

Overview of recent SCALE activities for Non-LWR inventory and decay heat analysis

In 2019, the US Nuclear Regulatory Commission initiated a project for the development and assessments of non-light-water reactor (non-LWR) accident progression using the SCALE and MELCOR simulation tools. SCALE simulations are used to generate nuclide inventories, full-core power distributions, decay heat, and kinetics parameters to initialize MELCOR simulations of severe accident scenarios. Five non-LWR concepts were studied: high-temperature gas-cooled reactor (HTGR), heat pipe reactor (HPR), high-temperature fluoride salt-cooled reactor (FHR), molten salt-fueled reactor (MSR), and sodium-cooled fast reactor (SFR). This paper summarizes the SCALE results obtained in 2021 for the first three non-LWR concepts, compares characteristics and results to common LWRs, and provides the strategy for the analysis of the remaining two non-LWRs. (authors)

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗

Expanding Activity Allocation Models to Daily Activities: Tracking Simulated Agent Trips to Exercise Locations in Clarksville, TN

Exercise facilities have been proven to have numerous physical, mental, and psychological benefits, yet exercise facilities are still inaccessible to a large portion of the population. This study serves to explore the accessibility of fitness centres through geographical, demographic, and temporal lenses through an expansion of the UrbanPop framework that seeks to allocate simulated agents to fitness centres in the Clarksville metro to explore the effects of travel distances on different demographics throughout the week. Findings indicate that senior and retired demographics consistently travel longer distances to exercise in the larger Clarksville area, likely due to tendencies to live further from the center of the metropolitan area. Furthermore, all demographics tend to travel further distances to exercise on the weekends rather than the weekdays, indicating that travel distance can affect likelihood of agents to travel, especially on weekdays when many agents are in the workforce or participating in schooling.

99 GENERAL AND MISCELLANEOUS↗

Isolation of Physiologically Active Thylakoids and Their Use in Energy-Dependent Protein Transport Assays

Chloroplasts are the organelles in green plants responsible for carrying out numerous essential metabolic pathways, most notably photosynthesis. Within the chloroplasts, the thylakoid membrane system houses all the photosynthetic pigments, reaction center complexes, and most of the electron carriers, and is responsible for light-dependent ATP synthesis. Over 90% of chloroplast proteins are encoded in the nucleus, translated in the cytosol, and subsequently imported into the chloroplast. Further protein transport into or across the thylakoid membrane utilizes one of four translocation pathways. Here, we describe a high-yield method for isolation of transport-competent thylakoids from peas (Pisum sativum), along with transport assays through the three energy-dependent cpTat, cpSec1, and cpSRP-mediated pathways. We report these methods enable experiments relating to thylakoid protein localization, transport energetics, and the mechanisms of protein translocation across biological membranes.

59 BASIC BIOLOGICAL SCIENCES↗

Drosophila Nanos acts as a molecular clamp that modulates the RNA-binding and repression activities of Pumilio

Collaboration among the multitude of RNA-binding proteins (RBPs) is ubiquitous, yet our understanding of these key regulatory complexes has been limited to single RBPs. We investigated combinatorial translational regulation by Drosophila Pumilio (Pum) and Nanos (Nos), which control development, fertility, and neuronal functions. Our results show how the specificity of one RBP (Pum) is modulated by cooperative RNA recognition with a second RBP (Nos) to synergistically repress mRNAs. Crystal structures of Nos-Pum-RNA complexes reveal that Nos embraces Pum and RNA, contributes sequence-specific contacts, and increases Pum RNA-binding affinity. Nos shifts the recognition sequence and promotes repression complex formation on mRNAs that are not stably bound by Pum alone, explaining the preponderance of sub-optimal Pum sites regulated in vivo . Our results illuminate the molecular mechanism of a regulatory switch controlling crucial gene expression programs, and provide a framework for understanding how the partnering of RBPs evokes changes in binding specificity that underlie regulatory network dynamics.

Weidmann, Chase A.↗