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Sample records for coelicolor development demonstrates

  1. Plasticity of Streptomyces coelicolor Membrane Composition Under Different Growth Conditions and During Development

    PubMed Central

    Sandoval-Calderón, Mario; Nguyen, Don D.; Kapono, Clifford A.; Herron, Paul; Dorrestein, Pieter C.; Sohlenkamp, Christian

    2015-01-01

    Streptomyces coelicolor is a model actinomycete that is well known for the diversity of its secondary metabolism and its complex life cycle. As a soil inhabitant, it is exposed to heterogeneous and frequently changing environmental circumstances. In the present work, we studied the effect of diverse growth conditions and phosphate depletion on its lipid profile and the relationship between membrane lipid composition and development in S. coelicolor. The lipid profile from cultures grown on solid media, which is closer to the natural habitat of this microorganism, does not resemble the previously reported lipid composition from liquid grown cultures of S. coelicolor. Wide variations were also observed across different media, growth phases, and developmental stages indicating active membrane remodeling. Ornithine lipids (OL) are phosphorus-free polar lipids that were accumulated mainly during sporulation stages, but were also major components of the membrane under phosphorus limitation. In contrast, phosphatidylethanolamine, which had been reported as one of the major polar lipids in the genus Streptomyces, is almost absent under these conditions. We identified one of the genes responsible for the synthesis of OL (SCO0921) and found that its inactivation causes the absence of OL, precocious morphological development and actinorhodin production. Our observations indicate a remarkable plasticity of the membrane composition in this bacterial species, reveal a higher metabolic complexity than expected, and suggest a relationship between cytoplasmic membrane components and the differentiation programs in S. coelicolor. PMID:26733994

  2. Transcriptomic Analysis of Liquid Non-Sporulating Streptomyces coelicolor Cultures Demonstrates the Existence of a Complex Differentiation Comparable to That Occurring in Solid Sporulating Cultures

    PubMed Central

    Yagüe, Paula; Rodríguez-García, Antonio; López-García, María Teresa; Rioseras, Beatriz; Martín, Juan Francisco; Sánchez, Jesús; Manteca, Angel

    2014-01-01

    Streptomyces species produce many clinically relevant secondary metabolites and exhibit a complex development that includes hyphal differentiation and sporulation in solid cultures. Industrial fermentations are usually performed in liquid cultures, conditions in which Streptomyces strains generally do not sporulate, and it was traditionally assumed that no differentiation took place. The aim of this work was to compare the transcriptomes of S. coelicolor growing in liquid and solid cultures, deepening the knowledge of Streptomyces differentiation. Microarrays demonstrated that gene expression in liquid and solid cultures were comparable and data indicated that physiological differentiation was similar for both conditions. Eighty-six percent of all transcripts showed similar abundances in liquid and solid cultures, such as those involved in the biosynthesis of actinorhodin (actVA, actII-4) and undecylprodigiosin (redF); activation of secondary metabolism (absR1, ndsA); genes regulating hydrophobic cover formation (aerial mycelium) (bldB, bldC, bldM, bldN, sapA, chpC, chpD, chpE, chpH, ramA, ramC, ramS); and even some genes regulating early stages of sporulation (wblA, whiG, whiH, whiJ). The two most important differences between transcriptomes from liquid and solid cultures were: first, genes related to secondary metabolite biosynthesis (CDA, CPK, coelichelin, desferrioxamine clusters) were highly up-regulated in liquid but not in solid cultures; and second, genes involved in the final stages of hydrophobic cover/spore maturation (chpF, rdlA, whiE, sfr) were up-regulated in solid but not in liquid cultures. New information was also provided for several non-characterized genes differentially expressed in liquid and solid cultures which might be regulating, at least in part, the metabolic and developmental differences observed between liquid and solid cultures. PMID:24466012

  3. Exploration Technology Development & Demonstration

    NASA Video Gallery

    Chris Moore delivers a presentation from the Exploration Technology Development & Demonstration (ETDD) study team on May 25, 2010, at the NASA Exploration Enterprise Workshop held in Galveston, TX....

  4. Mycelium differentiation and development of Streptomyces coelicolor in lab-scale bioreactors: Programmed cell death, differentiation, and lysis are closely linked to undecylprodigiosin and actinorhodin production

    PubMed Central

    Rioseras, Beatriz; López-García, María Teresa; Yagüe, Paula; Sánchez, Jesús; Manteca, Ángel

    2013-01-01

    Streptomycetes are mycelium-forming bacteria that produce two thirds of clinically relevant secondary metabolites. Secondary metabolite production is activated at specific developmental stages of Streptomyces life cycle. Despite this, Streptomyces differentiation in industrial bioreactors tends to be underestimated and the most important parameters managed are only indirectly related to differentiation: modifications to the culture media, optimization of productive strains by random or directed mutagenesis, analysis of biophysical parameters, etc. In this work the relationship between differentiation and antibiotic production in lab-scale bioreactors was defined. Streptomyces coelicolor was used as a model strain. Morphological differentiation was comparable to that occurring during pre-sporulation stages in solid cultures: an initial compartmentalized mycelium suffers a programmed cell death, and remaining viable segments then differentiate to a second multinucleated antibiotic-producing mycelium. Differentiation was demonstrated to be one of the keys to interpreting biophysical fermentation parameters and to rationalizing the optimization of secondary metabolite production in bioreactors. PMID:24240146

  5. Mycelium differentiation and development of Streptomyces coelicolor in lab-scale bioreactors: programmed cell death, differentiation, and lysis are closely linked to undecylprodigiosin and actinorhodin production.

    PubMed

    Rioseras, Beatriz; López-García, María Teresa; Yagüe, Paula; Sánchez, Jesús; Manteca, Angel

    2014-01-01

    Streptomycetes are mycelium-forming bacteria that produce two thirds of clinically relevant secondary metabolites. Secondary metabolite production is activated at specific developmental stages of Streptomyces life cycle. Despite this, Streptomyces differentiation in industrial bioreactors tends to be underestimated and the most important parameters managed are only indirectly related to differentiation: modifications to the culture media, optimization of productive strains by random or directed mutagenesis, analysis of biophysical parameters, etc. In this work the relationship between differentiation and antibiotic production in lab-scale bioreactors was defined. Streptomyces coelicolor was used as a model strain. Morphological differentiation was comparable to that occurring during pre-sporulation stages in solid cultures: an initial compartmentalized mycelium suffers a programmed cell death, and remaining viable segments then differentiate to a second multinucleated antibiotic-producing mycelium. Differentiation was demonstrated to be one of the keys to interpreting biophysical fermentation parameters and to rationalizing the optimization of secondary metabolite production in bioreactors. PMID:24240146

  6. Transcriptome dynamics-based operon prediction and verification in Streptomyces coelicolor

    PubMed Central

    Charaniya, Salim; Mehra, Sarika; Lian, Wei; Jayapal, Karthik P.; Karypis, George; Hu, Wei-Shou

    2007-01-01

    Streptomyces spp. produce a variety of valuable secondary metabolites, which are regulated in a spatio-temporal manner by a complex network of inter-connected gene products. Using a compilation of genome-scale temporal transcriptome data for the model organism, Streptomyces coelicolor, under different environmental and genetic perturbations, we have developed a supervised machine-learning method for operon prediction in this microorganism. We demonstrate that, using features dependent on transcriptome dynamics and genome sequence, a support vector machines (SVM)-based classification algorithm can accurately classify >90% of gene pairs in a set of known operons. Based on model predictions for the entire genome, we verified the co-transcription of more than 250 gene pairs by RT-PCR. These results vastly increase the database of known operons in S. coelicolor and provide valuable information for exploring gene function and regulation to harness the potential of this differentiating microorganism for synthesis of natural products. PMID:17959654

  7. Transcriptomics-based strain optimization tool for designing secondary metabolite overproducing strains of Streptomyces coelicolor.

    PubMed

    Kim, Minsuk; Yi, Jeong Sang; Lakshmanan, Meiyappan; Lee, Dong-Yup; Kim, Byung-Gee

    2016-03-01

    In silico model-driven analysis using genome-scale model of metabolism (GEM) has been recognized as a promising method for microbial strain improvement. However, most of the current GEM-based strain design algorithms based on flux balance analysis (FBA) heavily rely on the steady-state and optimality assumptions without considering any regulatory information. Thus, their practical usage is quite limited, especially in its application to secondary metabolites overproduction. In this study, we developed a transcriptomics-based strain optimization tool (tSOT) in order to overcome such limitations by integrating transcriptomic data into GEM. Initially, we evaluated existing algorithms for integrating transcriptomic data into GEM using Streptomyces coelicolor dataset, and identified iMAT algorithm as the only and the best algorithm for characterizing the secondary metabolism of S. coelicolor. Subsequently, we developed tSOT platform where iMAT is adopted to predict the reaction states, and successfully demonstrated its applicability to secondary metabolites overproduction by designing actinorhodin (ACT), a polyketide antibiotic, overproducing strain of S. coelicolor. Mutants overexpressing tSOT targets such as ribulose 5-phosphate 3-epimerase and NADP-dependent malic enzyme showed 2 and 1.8-fold increase in ACT production, thereby validating the tSOT prediction. It is expected that tSOT can be used for solving other metabolic engineering problems which could not be addressed by current strain design algorithms, especially for the secondary metabolite overproductions. PMID:26369755

  8. A Laterally Acquired Galactose Oxidase-Like Gene Is Required for Aerial Development during Osmotic Stress in Streptomyces coelicolor

    PubMed Central

    Liman, Recep; Facey, Paul D.; van Keulen, Geertje; Dyson, Paul J.; Del Sol, Ricardo

    2013-01-01

    Phylogenetic reconstruction revealed that most Actinobacterial orthologs of S. coelicolor SCO2837, encoding a metal-dependent galactose oxidase-like protein, are found within Streptomyces and were probably acquired by horizontal gene transfer from fungi. Disruption of SCO2837 (glxA) caused a conditional bld phenotype that could not be reversed by extracellular complementation. Studies aimed at characterising the regulation of expression of glxA showed that it is not a target for other bld genes. We provide evidence that glxA is required for osmotic adaptation, although independently from the known osmotic stress response element SigB. glxA has been predicted to be part of an operon with the transcription unit comprising the upstream cslA gene and glxA. However, both phenotypic and expression studies indicate that it is also expressed from an independent promoter region internal to cslA. GlxA displays an in situ localisation pattern similar to that one observed for CslA at hyphal tips, but localisation of the former is independent of the latter. The functional role of GlxA in relation to CslA is discussed. PMID:23326581

  9. Phage-mediated cloning of bldA, a region involved in Streptomyces coelicolor morphological development, and its analysis by genetic complementation.

    PubMed Central

    Piret, J M; Chater, K F

    1985-01-01

    Streptomyces coelicolor bald (bld) mutants form colonies of vegetative substrate mycelium, but do not develop aerial hyphae or spore chains. The bldA strains form none of the four antibiotics known to be produced by the parent strain. With a vector derived from the temperate bacteriophage phi C31, a 5.6-kilobase fragment of wildtype DNA was cloned which restored sporulation to five independent bldA mutants when lysogenized with the recombinant phage. The cloned gene(s) was dominant over the mutant alleles. Phage integration by recombination of the cloned bldA+ DNA with the bldA region of each mutant produced mainly sporulating colonies, presumably heterozygous bldA+/bldA partial diploids for the insert DNA. However, a minority of these primary transductants were bald and were apparently homozygous bldA/bldA mutant partial diploids, formed by some homogenetization process. The phages released from the bald lysogens carried bldA mutations and were used to show that bldA+ sequences had been cloned and that fine mapping of the region could be performed. Images PMID:2993254

  10. Cyclic Di-GMP Phosphodiesterases RmdA and RmdB Are Involved in Regulating Colony Morphology and Development in Streptomyces coelicolor

    PubMed Central

    Hull, Travis D.; Ryu, Min-Hyung; Sullivan, Matthew J.; Johnson, Ryan C.; Klena, Nikolai T.; Geiger, Robert M.; Gomelsky, Mark

    2012-01-01

    Cyclic dimeric GMP (c-di-GMP) regulates numerous processes in Gram-negative bacteria, yet little is known about its role in Gram-positive bacteria. Here we characterize two c-di-GMP phosphodiesterases from the filamentous high-GC Gram-positive actinobacterium Streptomyces coelicolor, involved in controlling colony morphology and development. A transposon mutation in one of the two phosphodiesterase genes, SCO0928, hereby designated rmdA (regulator of morphology and development A), resulted in decreased levels of spore-specific gray pigment and a delay in spore formation. The RmdA protein contains GGDEF-EAL domains arranged in tandem and possesses c-di-GMP phosphodiesterase activity, as is evident from in vitro enzymatic assays using the purified protein. RmdA contains a PAS9 domain and is a hemoprotein. Inactivation of another GGDEF-EAL-encoding gene, SCO5495, designated rmdB, resulted in a phenotype identical to that of the rmdA mutant. Purified soluble fragment of RmdB devoid of transmembrane domains also possesses c-di-GMP phosphodiesterase activity. The rmdA rmdB double mutant has a bald phenotype and is impaired in aerial mycelium formation. This suggests that RmdA and RmdB functions are additive and at least partially overlapping. The rmdA and rmdB mutations likely result in increased local pools of intracellular c-di-GMP, because intracellular c-di-GMP levels in the single mutants did not differ significantly from those of the wild type, whereas in the double rmdA rmdB mutant, c-di-GMP levels were 3-fold higher than those in the wild type. This study highlights the importance of c-di-GMP-dependent signaling in actinomycete colony morphology and development and identifies two c-di-GMP phosphodiesterases controlling these processes. PMID:22753061

  11. Regulation of a nickel-cobalt efflux system and nickel homeostasis in a soil actinobacterium Streptomyces coelicolor.

    PubMed

    Kim, Hae Mi; Ahn, Bo-Eun; Lee, Ju-Hyung; Roe, Jung-Hye

    2015-04-01

    In Streptomyces coelicolor, a soil actinobacterium capable of morphological differentiation and complex secondary metabolism, nickel deficiency is sensed by Nur, a Ni-specific Fur family regulator that controls nickel uptake systems (NikABCDE and NikMNOQ) and both Fe-containing and Ni-containing superoxide dismutases (SodF and SodN). On the other hand, the nickel efflux system and its regulator have not been elucidated. In this study, we demonstrate that an ArsR/SmtB family metalloregulator NmtR, a close homologue of NmtR from Mycobacterium tuberculosis, controls a putative efflux pump of P1-type ATPase (NmtA) in S. coelicolor. NmtR binds to the nmtA promoter region to repress its transcription, and is dissociated in the presence of Ni(ii) and Co(ii). Disruption of the nmtA gene makes cells more sensitive to nickel and cobalt, consistent with its predicted role in encoding a Ni-Co-efflux pump. Growth of S. coelicolor in complex YEME medium is only marginally inhibited by up to 0.5 mM Ni(ii), with significant growth retardation at 1 mM. Nur-regulated sodF and nikA genes are repressed at less than 0.1 μM added NiSO4 whereas NmtR-regulated nmtA transcription is induced at 0.5 mM or more Ni(ii). This reveals the extreme sensitivity of S. coelicolor to nickel deficiency as well as tolerance to surplus nickel. How this organism and possibly other actinomycetes have evolved to develop such a highly Ni-tolerant physiology and how the highly sensitive regulator Nur and the obtuse regulator NmtR achieve their characteristic Ni-sensitivity are interesting questions to solve in the future. PMID:25697558

  12. New Knowledge from Old: In silico discovery of novel protein domains in Streptomyces coelicolor

    PubMed Central

    Yeats, Corin; Bentley, Stephen; Bateman, Alex

    2003-01-01

    Background Streptomyces coelicolor has long been considered a remarkable bacterium with a complex life-cycle, ubiquitous environmental distribution, linear chromosomes and plasmids, and a huge range of pharmaceutically useful secondary metabolites. Completion of the genome sequence demonstrated that this diversity carried through to the genetic level, with over 7000 genes identified. We sought to expand our understanding of this organism at the molecular level through identification and annotation of novel protein domains. Protein domains are the evolutionary conserved units from which proteins are formed. Results Two automated methods were employed to rapidly generate an optimised set of targets, which were subsequently analysed manually. A final set of 37 domains or structural repeats, represented 204 times in the genome, was developed. Using these families enabled us to correlate items of information from many different resources. Several immediately enhance our understanding both of S. coelicolor and also general bacterial molecular mechanisms, including cell wall biosynthesis regulation and streptomycete telomere maintenance. Discussion Delineation of protein domain families enables detailed analysis of protein function, as well as identification of likely regions or residues of particular interest. Hence this kind of prior approach can increase the rate of discovery in the laboratory. Furthermore we demonstrate that using this type of in silico method it is possible to fairly rapidly generate new biological information from previously uncorrelated data. PMID:12625841

  13. Cell immobilization of Streptomyces coelicolor: effect on differentiation and actinorhodin production

    PubMed Central

    López-García, María Teresa; Rioseras, Beatriz; Yagüe, Paula; Álvarez, José Ramón; Manteca, Ángel

    2015-01-01

    Summary Streptomycetes are mycelium-forming bacteria that produce two thirds of the clinically relevant secondary metabolites. Despite the fact that secondary metabolite production is activated at specific developmental stages of the Streptomyces spp. life cycle, different streptomycetes show different behaviors, and fermentation conditions need to be optimized for each specific strain and secondary metabolite. Cell-encapsulation constitutes an interesting alternative to classical fermentations, which was demonstrated to be useful in Streptomyces, but development under these conditions remained unexplored. In this work, the influence of cell-encapsulation in hyphae differentiation and actinorhodin production was explored in the model Streptomyces coelicolor strain. Encapsulation led to a delay in growth and to a reduction of mycelium density and cell death. The high proportion of viable hyphae duplicated extracellular actinorhodin production in the encapsulated cultures with respect to the non-encapsulated ones. PMID:26418851

  14. TOF-SIMS investigation of Streptomyces coelicolor, a mycelial bacterium

    NASA Astrophysics Data System (ADS)

    Vaidyanathan, Seetharaman; Fletcher, John S.; Lockyer, Nicholas P.; Vickerman, John C.

    2008-12-01

    Streptomyces coelicolor is a mycelial microorganism that produces several secondary metabolites, including antibiotics. The physiology of the organism has largely been investigated in liquid cultures due to ease of monitoring different physiological parameters and more homogeneous culture conditions. However, solid cultures reflect the natural physiology of the microorganism better, given that in its natural state it grows in the soil. Imaging mass spectrometry with TOF-SIMS and C 60+ primary ion beams offers a potential route to studying chemical changes at the molecular level, both intracellular and extracellular that can help in understanding the natural physiology of the microorganism. Here, we report the application of the technique for studying the lateral distribution of the chemical species detected in a population, grown in both liquid and solid cultures. The capability of the technique for studying biological systems with minimal system intervention is demonstrated.

  15. Codes & standards research, development & demonstration Roadmap

    SciTech Connect

    None, None

    2008-07-22

    This Roadmap is a guide to the Research, Development & Demonstration activities that will provide data required for SDOs to develop performance-based codes and standards for a commercial hydrogen fueled transportation sector in the U.S.

  16. The Absence of Pupylation (Prokaryotic Ubiquitin-Like Protein Modification) Affects Morphological and Physiological Differentiation in Streptomyces coelicolor

    PubMed Central

    Seghezzi, Nicolas; Duchateau, Magalie; Gominet, Myriam; Kofroňová, Olga; Benada, Oldřich; Mazodier, Philippe

    2015-01-01

    ABSTRACT Protein turnover is essential in all living organisms for the maintenance of normal cell physiology. In eukaryotes, most cellular protein turnover involves the ubiquitin-proteasome pathway, in which proteins tagged with ubiquitin are targeted to the proteasome for degradation. In contrast, most bacteria lack a proteasome but harbor proteases for protein turnover. However, some actinobacteria, such as mycobacteria, possess a proteasome in addition to these proteases. A prokaryotic ubiquitination-like tagging process in mycobacteria was described and was named pupylation: proteins are tagged with Pup (prokaryotic ubiquitin-like protein) and directed to the proteasome for degradation. We report pupylation in another actinobacterium, Streptomyces coelicolor. Both the morphology and life cycle of Streptomyces species are complex (formation of a substrate and aerial mycelium followed by sporulation), and these bacteria are prolific producers of secondary metabolites with important medicinal and agricultural applications. The genes encoding the pupylation system in S. coelicolor are expressed at various stages of development. We demonstrated that pupylation targets numerous proteins and identified 20 of them. Furthermore, we established that abolition of pupylation has substantial effects on morphological and metabolic differentiation and on resistance to oxidative stress. In contrast, in most cases, a proteasome-deficient mutant showed only modest perturbations under the same conditions. Thus, the phenotype of the pup mutant does not appear to be due solely to defective proteasomal degradation. Presumably, pupylation has roles in addition to directing proteins to the proteasome. IMPORTANCE Streptomyces spp. are filamentous and sporulating actinobacteria, remarkable for their morphological and metabolic differentiation. They produce numerous bioactive compounds, including antifungal, antibiotic, and antitumor compounds. There is therefore considerable interest in

  17. Development and Demonstration of Ultrafiltration Simulants

    SciTech Connect

    Russell, Renee L.; Billing, Justin M.; Peterson, Reid A.; Rinehart, Donald E.; Smith, Harry D.

    2009-02-24

    According to Bechtel National, Inc. (BNI) Test Specification 24590-PTF-TSP-RT-06-006, Rev 0, Simulant Development to Support the Development and Demonstration of Leaching and Ultrafiltration Pretreatment Processes,” simulants for boehmite, gibbsite, and filtration are to be developed that can be used in subsequent bench and integrated testing of the leaching/filtration processes for the waste treatment plant (WTP). These simulants will then be used to demonstrate the leaching process and to help refine processing conditions which may impact safety basis considerations (Smith 2006). This report documents the results of the filtration simulant development.

  18. The Wisconsin Rural Development Demonstration Project.

    ERIC Educational Resources Information Center

    McCoy, Palmer E.; Jones, Eleanor

    Nineteen Wisconsin counties were the site of the four-year Rural Development Demonstration Project, one of five such projects in the nation. Among the project's objectives were to accelerate sound economic, social, cultural and environmental development and to provide alternatives for living in non-metropolitan areas while providing greater…

  19. The Level of AdpA Directly Affects Expression of Developmental Genes in Streptomyces coelicolor ▿ †

    PubMed Central

    Wolański, Marcin; Donczew, Rafał; Kois-Ostrowska, Agnieszka; Masiewicz, Paweł; Jakimowicz, Dagmara; Zakrzewska-Czerwińska, Jolanta

    2011-01-01

    AdpA is a key regulator of morphological differentiation in Streptomyces. In contrast to Streptomyces griseus, relatively little is known about AdpA protein functions in Streptomyces coelicolor. Here, we report for the first time the translation accumulation profile of the S. coelicolor adpA (adpASc) gene; the level of S. coelicolor AdpA (AdpASc) increased, reaching a maximum in the early stage of aerial mycelium formation (after 36 h), and remained relatively stable for the next several hours (48 to 60 h), and then the signal intensity decreased considerably. AdpASc specifically binds the adpASc promoter region in vitro and in vivo, suggesting that its expression is autoregulated; surprisingly, in contrast to S. griseus, the protein presumably acts as a transcriptional activator. We also demonstrate a direct influence of AdpASc on the expression of several genes whose products play key roles in the differentiation of S. coelicolor: STI, a protease inhibitor; RamR, an atypical response regulator that itself activates expression of the genes for a small modified peptide that is required for aerial growth; and ClpP1, an ATP-dependent protease. The diverse influence of AdpASc protein on the expression of the analyzed genes presumably results mainly from different affinities of AdpASc protein to individual promoters. PMID:21926228

  20. Integrated Metabolomics Approach Facilitates Discovery of an Unpredicted Natural Product Suite from Streptomyces coelicolor M145

    PubMed Central

    Sidebottom, Ashley M.; Johnson, Andrew R.; Karty, Jonathan A.; Trader, Darci J.; Carlson, Erin E.

    2013-01-01

    Natural products exhibit a broad range of biological properties and have been a crucial source of therapeutic agents and novel scaffolds. Although bacterial secondary metabolomes are widely explored, they remain incompletely cataloged by current isolation and characterization strategies. To identify metabolites residing in unexplored chemical space, we have developed an integrated discovery approach that combines bacterial growth perturbation, accurate mass spectrometry, comparative mass spectra data analysis, and fragmentation spectra clustering for the identification of low-abundant, novel compounds from complex biological matrices. In this investigation, we analyzed the secreted metabolome of the extensively studied Actinomycete, Streptomyces coelicolor M145, and discovered a low-abundant suite of 15 tri-hydroxamate, amphiphilic siderophores. Compounds in this class have primarily been observed in marine microorganisms making their detection in the soil-dwelling S. coelicolor M145 significant. At least ten of these ferrioxamine-based molecules are not known to be produced by any organism and none have previously been detected from S. coelicolor M145. In addition, we confirmed the production of ferrioxamine D1, a relatively hydrophilic family member that has not been shown to be biosynthesized by this organism. The identified molecules are part of only a small list of secondary metabolites that have been discovered since sequencing of S. coelicolor M145 revealed that it possessed numerous putative secondary metabolite-producing gene clusters with no known metabolites. Thus, the identified siderophores represent the unexplored metabolic potential of both well-studied and new organisms that could be uncovered with our sensitive and robust approach. PMID:23777274

  1. Bioinformatic identification of novel regulatory DNA sequence motifs in Streptomyces coelicolor

    PubMed Central

    Studholme, David J; Bentley, Stephen D; Kormanec, Jan

    2004-01-01

    Background Streptomyces coelicolor is a bacterium with a vast repertoire of metabolic functions and complex systems of cellular development. Its genome sequence is rich in genes that encode regulatory proteins to control these processes in response to its changing environment. We wished to apply a recently published bioinformatic method for identifying novel regulatory sequence signals to gain new insights into regulation in S. coelicolor. Results The method involved production of position-specific weight matrices from alignments of over-represented words of DNA sequence. We generated 2497 weight matrices, each representing a candidate regulatory DNA sequence motif. We scanned the genome sequence of S. coelicolor against each of these matrices. A DNA sequence motif represented by one of the matrices was found preferentially in non-coding sequences immediately upstream of genes involved in polysaccharide degradation, including several that encode chitinases. This motif (TGGTCTAGACCA) was also found upstream of genes encoding components of the phosphoenolpyruvate phosphotransfer system (PTS). We hypothesise that this DNA sequence motif represents a regulatory element that is responsive to availability of carbon-sources. Other motifs of potential biological significance were found upstream of genes implicated in secondary metabolism (TTAGGTtAGgCTaACCTAA), sigma factors (TGACN19TGAC), DNA replication and repair (ttgtCAGTGN13TGGA), nucleotide conversions (CTACgcNCGTAG), and ArsR (TCAGN12TCAG). A motif found upstream of genes involved in chromosome replication (TGTCagtgcN7Tagg) was similar to a previously described motif found in UV-responsive promoters. Conclusions We successfully applied a recently published in silico method to identify conserved sequence motifs in S. coelicolor that may be biologically significant as regulatory elements. Our data are broadly consistent with and further extend data from previously published studies. We invite experimental testing of

  2. Human Support Technology Research, Development and Demonstration

    NASA Technical Reports Server (NTRS)

    Joshi, Jitendra; Trinh, Eugene

    2004-01-01

    The Human Support Technology research, development, and demonstration program address es the following areas at TRL: Advanced Power and Propulsion. Cryogenic fluid management. Closed-loop life support and Habitability. Extravehicular activity systems. Scientific data collection and analysis. and Planetary in-situ resource utilization.

  3. Repression of Antibiotic Production and Sporulation in Streptomyces coelicolor by Overexpression of a TetR Family Transcriptional Regulator ▿ †

    PubMed Central

    Xu, Delin; Seghezzi, Nicolas; Esnault, Catherine; Virolle, Marie-Joelle

    2010-01-01

    The overexpression of a regulatory gene of the TetR family (SCO3201) originating either from Streptomyces lividans or from Streptomyces coelicolor was shown to strongly repress antibiotic production (calcium-dependent antibiotic [CDA], undecylprodigiosin [RED], and actinorhodin [ACT]) of S. coelicolor and of the ppk mutant strain of S. lividans. Curiously, the overexpression of this gene also had a strong inhibitory effect on the sporulation process of S. coelicolor but not on that of S. lividans. SCO3201 was shown to negatively regulate its own transcription, and its DNA binding motif was found to overlap its −35 promoter sequence. The interruption of this gene in S. lividans or S. coelicolor did not lead to any obvious phenotypes, indicating that when overexpressed SCO3201 likely controls the expression of target genes of other TetR regulators involved in the regulation of the metabolic and morphological differentiation process in S. coelicolor. The direct and functional interaction of SCO3201 with the promoter region of scbA, a gene under the positive control of the TetR-like regulator, ScbR, was indeed demonstrated by in vitro as well as in vivo approaches. PMID:20935121

  4. Integrated powerhead demonstration full flow cycle development

    NASA Astrophysics Data System (ADS)

    Jones, J. Mathew; Nichols, James T.; Sack, William F.; Boyce, William D.; Hayes, William A.

    1998-01-01

    The Integrated Powerhead Demonstration (IPD) is a 1,112,000 N (250,000 lbf) thrust (at sea level) LOX/LH2 demonstration of a full flow cycle in an integrated system configuration. Aerojet and Rocketdyne are on contract to the Air Force Research Laboratory to design, develop, and deliver the required components, and to provide test support to accomplish the demonstration. Rocketdyne is on contract to provide a fuel and oxygen turbopump, a gas-gas injector, and system engineering and integration. Aerojet is on contract to provide a fuel and oxygen preburner, a main combustion chamber, and a nozzle. The IPD components are being designed with Military Spaceplane (MSP) performance and operability requirements in mind. These requirements include: lifetime >=200 missions, mean time between overhauls >=100 cycles, and a capability to throttle from 20% to 100% of full power. These requirements bring new challenges both in designing and testing the components. This paper will provide some insight into these issues. Lessons learned from operating and supporting the space shuttle main engine (SSME) have been reviewed and incorporated where applicable. The IPD program will demonstrate phase I goals of the Integrated High Payoff Rocket Propulsion Technology (IHPRPT) program while demonstrating key propulsion technologies that will be available for MSP concepts. The demonstration will take place on Test Stand 2A at the Air Force Research Laboratory at Edwards AFB. The component tests will begin in 1999 and the integrated system tests will be completed in 2002.

  5. Hydrogen fueling station development and demonstration

    SciTech Connect

    Edeskuty, F.J.; Daney, D.; Daugherty, M.; Hill, D.; Prenger, F.C.

    1996-09-01

    This is the final report of a one-year, Laboratory-Directed Research and Development (LDRD) project at the Los Alamos National Laboratory (LANL). This project sought to develop and demonstrate a hydrogen fueling station for vehicles. Such stations are an essential infrastructural element in the practical application of hydrogen as vehicle fuel, and a number of issues such as safety, efficiency, design, and operating procedures can only be accurately addressed by a practical demonstration. Regardless of whether the vehicle is powered by an internal combustion engine or fuel cell, or whether the vehicle has a liquid or gaseous fuel tank, the fueling station is a critical technology that is the link between the local storage facility and the vehicle.

  6. Servicer system demonstration plan and capability development

    NASA Technical Reports Server (NTRS)

    1987-01-01

    An orbital maneuvering vehicle (OMV) front end kit is defined which is capable of performing in-situ fluid resupply and modular maintenance of free flying spacecraft based on the integrated orbital servicing system (IOSS) concept. The compatibility of the IOSS to perform gas and fluid umbilical connect and disconnect functions utilizing connect systems currently available or in development is addressed. A series of tasks involving on-orbit servicing and the engineering test unit (ETU) of the on-orbit service were studied. The objective is the advancement of orbital servicing by expanding the Spacecraft Servicing Demonstration Plan (SSDP) to include detail demonstration planning using the Multimission Modular Spacecraft (MMS) and upgrading the ETU control.

  7. Wireless Power Transmission Technology Development and Demonstrations

    NASA Astrophysics Data System (ADS)

    Steinsiek, F.; Weber, K.-H.; Foth, W.-P.; Foth, H. J.; Schäfer, C.

    2004-12-01

    The Wireless Power Transmission (WPT) technology has been treated to a wide extent in the recent years. A broad variety of applications has been investigated, from earth to orbit, orbit to earth, in-orbit and planetary ones, as for moon and Mars missions. In this course the question to use laser or microwave technology has widely been discussed. Beaming energy to spacecrafts could provide an important space mission-economic potential. It promises significant reduction in the cost of access to space, for scientific and commercial missions, and increases the mission capabilities for in-space systems. For the future enhancement of ISS capabilities and operational efficiency, the use of WPT technology became part of the technology research planning for the ISS. The WPT may have the potential of providing operational benefits, increase of spacecraft systems efficiency for elements like co-orbiting platforms, transfer vehicles or other ISS related in-orbit spacecrafts, and planetary exploration vehicles. The laser technology provides specific technical, operational and economic benefits compared to microwave applications and provides the actual basis for the envisioned wireless power transmission concepts. An outlook in terms of future wireless power perspectives, both for terrestrial as for space-to-space scenarios is given; these applications are part of a technology demonstration roadmap for wireless power transmission key- and supporting technologies, which is characterized by dedicated technology demonstration milestones on ground and in space. The actual technology development philosophy as conceived at EADS-Space Transportation is described and includes main system demonstration missions, as a laboratory test bed employing a small rover system, a scaled airship model demonstration as planned in 2004 and an experiment onboard the International Space Station ISS. These demonstrations represent milestones in terms of technical capability verification on the way to

  8. Cardiolipin synthase is required for Streptomyces coelicolor morphogenesis

    PubMed Central

    Jyothikumar, Vinod; Klanbut, Khanungkan; Tiong, John; Roxburgh, James S.; Hunter, Iain S.; Smith, Terry K.; Herron, Paul R.

    2013-01-01

    Summary The fluid mosaic model has recently been amended to account for the existence of membrane domains enriched in certain phospholipids. In rod-shaped bacteria, the anionic phospholipid cardiolipin is enriched at the cell poles but its role in the morphogenesis of the filamentous bacterium Streptomyces coelicolor is unknown. It was impossible to delete clsA (cardiolipin synthase; SCO1389) unless complemented by a second copy of clsA elsewhere in the chromosome. When placed under the control of an inducible promoter, clsA expression, phospholipid profile and morphogenesis became inducer dependent. TLC analysis of phospholipid showed altered profiles upon depletion of clsA expression. Analysis of cardiolipin by mass spectrometry showed two distinct cardiolipin envelopes that reflected differences in acyl chain length; the level of the larger cardiolipin envelope was reduced in concert with clsA expression. ClsA-EGFP did not localize to specific locations, but cardiolipin itself showed enrichment at hyphal tips, branch points and anucleate regions. Quantitative analysis of hyphal dimensions showed that the mycelial architecture and the erection of aerial hyphae were affected by the expression of clsA. Overexpression of clsA resulted in weakened hyphal tips, misshaped aerial hyphae and anucleate spores and demonstrates that cardiolipin synthesis is a requirement for morphogenesis in Streptomyces. PMID:22409773

  9. Fuel consolidation demonstration: Consolidation concept development

    SciTech Connect

    Not Available

    1990-02-01

    EPRI, Northeast utilities Service Company (NUSCO), DOE, Baltimore Gas Electric Company, and Combustion Engineering, Inc. (C-E) are engaged in a program to develop a system for consolidating spent fuel, in which the consolidated fuel will be licensable by NRC for storage in the spent-fuel storage pool. Fuel consolidation offers a means of substantially increasing the capacity of spent-fuel storage pools. Consolidation equipment design, development, construction, and testing are being performed by C-E in Windsor, Connecticut. Seismic and structural evaluation of the capability of the Millstone Unit 2 spent-fuel pool and building to accommodate the increased fuel capacity is being conducted by NUSCO. NUSCO plans to obtain a license to store consolidated fuel in the Millstone-2 spent-fuel storage pool. NUSCO also plans to perform a hot demonstration of the integrated consolidation system with spent fuel at Millstone-2. This report describes the consolidation system design that forms the basis for the detailed design of the equipment comprising the system, including information on the fabrication and testing of the equipment. Appendix B describes an evaluation of the ability of the system under development to consolidate LWR spent-fuel assemblies other than the 14 {times} 14 fuel of C-E design stored at Millstone-2. A comparison was made of fuel-assembly designs on the basis of information available in open literature. It was concluded that with appropriate dimensional modifications the spent-fuel consolidation system equipment design is applicable to almost all PWR fuel-assembly configurations. 8 refs., 20 figs.

  10. Multiyear Research, Development and Demonstration Plan

    SciTech Connect

    EERE

    2012-03-16

    The Fuel Cell Technologies Office Multi-Year Research, Development, and Demonstration (MYRD&D) Plan describes the goals, objectives, technical targets, tasks, and schedules for all activities within the Fuel Cell Technologies (FCT) Office, which is part of the U.S. Department of Energy's Office of Energy Efficiency and Renewable Energy. The MYRD&D Plan is a living document, which is revised periodically to reflect progress in the technologies, revisions to developmental timelines and targets, updates based on external reviews, and changes in the scope of the FCT Office. The document was first published in 2003, and revised in 2005. The document was significantly revised in 2012 to reflect scientific advancements and the changing technological landscape. Any revisions made to the MYRD&D Plan are conducted through a rigorous Change Control process as documented in the Systems Integration section of this report. The hydrogen and fuel cell activities within DOE continue to receive extensive review by stakeholders in the hydrogen and fuel cell community, including panels of the National Research Council and the National Academy of Engineering.

  11. Servicers system demonstration plan and capability development

    NASA Technical Reports Server (NTRS)

    Bulboaca, M. A.; Cuseo, J. A.; Derocher, W. L., Jr.; Maples, R. W.; Reynolds, P. C.; Sterrett, R. A.

    1985-01-01

    A plan for the demonstration of the exchange of Multi-Mission Modular Spacecraft (MMS) modules using the servicer mechanism Engineering Test Unit (ETU) was prepared and executed. The plan included: establishment of requirements, conceptual design, selection of MMS spacecraft mockup configuration, selection of MMS module mockup configuration, evaluation of adequacy of ETU load capability, and selection of a stowage rack arrangement. The MMS module exchange demonstration mockup equipment was designed, fabricated, checked out, shipped, installed, and demonstrated.

  12. Interspecies Interactions Stimulate Diversification of the Streptomyces coelicolor Secreted Metabolome

    PubMed Central

    Traxler, Matthew F.; Watrous, Jeramie D.; Alexandrov, Theodore; Dorrestein, Pieter C.; Kolter, Roberto

    2013-01-01

    ABSTRACT Soils host diverse microbial communities that include filamentous actinobacteria (actinomycetes). These bacteria have been a rich source of useful metabolites, including antimicrobials, antifungals, anticancer agents, siderophores, and immunosuppressants. While humans have long exploited these compounds for therapeutic purposes, the role these natural products may play in mediating interactions between actinomycetes has been difficult to ascertain. As an initial step toward understanding these chemical interactions at a systems level, we employed the emerging techniques of nanospray desorption electrospray ionization (NanoDESI) and matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) imaging mass spectrometry to gain a global chemical view of the model bacterium Streptomyces coelicolor interacting with five other actinomycetes. In each interaction, the majority of secreted compounds associated with S. coelicolor colonies were unique, suggesting an idiosyncratic response from S. coelicolor. Spectral networking revealed a family of unknown compounds produced by S. coelicolor during several interactions. These compounds constitute an extended suite of at least 12 different desferrioxamines with acyl side chains of various lengths; their production was triggered by siderophores made by neighboring strains. Taken together, these results illustrate that chemical interactions between actinomycete bacteria exhibit high complexity and specificity and can drive differential secondary metabolite production. PMID:23963177

  13. Geothermal Energy Research Development and Demonstration Program

    SciTech Connect

    Not Available

    1980-06-01

    The Federal program's goal, strategy, plans, and achievements are summarized. In addition, geothermal development by state and local governments and, where available, by the private sector is described. (MHR)

  14. Game Changing Development Demonstration Video 2014

    NASA Video Gallery

    The Game Changing Development Program is a part of NASA’s Space Technology Mission Directorate. The Program advances space technologies that may lead to entirely new approaches for the Agency’s fut...

  15. Novel Two-Component Systems Implied in Antibiotic Production in Streptomyces coelicolor

    PubMed Central

    Yepes, Ana; Rico, Sergio; Rodríguez-García, Antonio; Santamaría, Ramón I.; Díaz, Margarita

    2011-01-01

    The abundance of two-component systems (TCSs) in Streptomyces coelicolor A3(2) genome indicates their importance in the physiology of this soil bacteria. Currently, several TCSs have been related to antibiotic regulation, and the purpose in this study was the characterization of five TCSs, selected by sequence homology with the well-known absA1A2 system, that could also be associated with this important process. Null mutants of the five TCSs were obtained and two mutants (ΔSCO1744/1745 and ΔSCO4596/4597/4598) showed significant differences in both antibiotic production and morphological differentiation, and have been renamed as abr (antibiotic regulator). No detectable changes in antibiotic production were found in the mutants in the systems that include the ORFs SCO3638/3639, SCO3640/3641 and SCO2165/2166 in any of the culture conditions assayed. The system SCO1744/1745 (AbrA1/A2) was involved in negative regulation of antibiotic production, and acted also as a negative regulator of the morphological differentiation. By contrast, the system SCO4596/4597/4598 (AbrC1/C2/C3), composed of two histidine kinases and one response regulator, had positive effects on both morphological development and antibiotic production. Microarray analyses of the ΔabrC1/C2/C3 and wild-type transcriptomes revealed downregulation of actII-ORF4 and cdaR genes, the actinorhodin and calcium-dependent antibiotic pathway-specific regulators respectively. These results demonstrated the involvement of these new two-component systems in antibiotic production and morphological differentiation by different approaches. One is a pleiotropic negative regulator: abrA1/A2. The other one is a positive regulator composed of three elements, two histidine kinases and one response regulator: abrC1/C2/C3. PMID:21625497

  16. Demonstration and Validation Assets: User Manual Development

    SciTech Connect

    2008-06-30

    This report documents the development of a database-supported user manual for DEMVAL assets in the NSTI area of operations and focuses on providing comprehensive user information on DEMVAL assets serving businesses with national security technology applications in southern New Mexico. The DEMVAL asset program is being developed as part of the NSPP, funded by both Department of Energy (DOE) and NNSA. This report describes the development of a comprehensive user manual system for delivering indexed DEMVAL asset information to be used in marketing and visibility materials and to NSTI clients, prospective clients, stakeholders, and any person or organization seeking it. The data about area DEMVAL asset providers are organized in an SQL database with updateable application structure that optimizes ease of access and customizes search ability for the user.

  17. Status of HTGR technology development and demonstration

    SciTech Connect

    Trauger, D.B.

    1983-01-01

    Several HGTR items which require thorough testing include the core support and pads, vessel insulation and liner, and graphite fuel blocks. Tests which are to be conducted in the Component Flow Test Loop are discussed. The neutron streaming experiment at the Tower Shielding Facility is described. Materials and fuels development studies are also discussed. (DLC)

  18. Fundamental role of cobalamin biosynthesis in the developmental growth of Streptomyces coelicolor A3 (2).

    PubMed

    Takano, Hideaki; Hagiwara, Kenta; Ueda, Kenji

    2015-03-01

    Cobalamin (Cbl) (synonym, vitamin B12) is the cobalt-containing cofactor produced only by some prokaryotes. Streptomyces is an effective Cbl producer. To study the role of Cbl production in Streptomyces, a knockout mutant for Cbl biosynthesis (cob) was generated in Streptomyces coelicolor A3 (2). The growth of the mutant was similar to that of the wild type in a rich medium, but inhibited in minimal medium, suggesting the involvement of Cbl in some step of primary metabolism. Methionine synthesis catalyzed by MetH, the Cbl-dependent methionine synthase, is a candidate. However, supplementing the minimal medium with methionine did not rescue the growth of the cob mutant, indicating that the availability of Cbl affects another primary function. Transcriptional analysis confirmed that the mutant induced metE encoding an alternative Cbl-independent methionine synthase, probably due to the Cbl-dependent riboswitch mechanism. The cob mutant produced low levels of pigment antibiotics and formed fewer aerial mycelium and spores in a rich medium, suggesting that a Cbl-dependent mechanism controls development. A similar developmental defect was observed for a knockout mutant for SCO4800, encoding the putative Cbl-dependent isobutyryl-CoA mutase (Icm) small subunit. Since the knockout of the Icm large subunit (SCO5415) did not affect the developmental phenotype, SCO4800 likely regulates development independently from SCO5415. Effective Cbl production is fundamental to the diverse functions underlying the complex developmental life cycle of S. coelicolor A3 (2). PMID:25547841

  19. A versatile PCR-based tandem epitope tagging system for Streptomyces coelicolor genome.

    PubMed

    Kim, Ji-Nu; Yi, Jeong Sang; Lee, Bo-Rahm; Kim, Eun-Jung; Kim, Min Woo; Song, Yoseb; Cho, Byung-Kwan; Kim, Byung-Gee

    2012-07-20

    Epitope tagging approaches have been widely used for the analysis of functions, interactions and subcellular distributions of proteins. However, incorporating epitope sequence into protein loci in Streptomyces is time-consuming procedure due to the absence of the versatile tagging methods. Here, we developed a versatile PCR-based tandem epitope tagging tool for the Streptomyces genome engineering. We constructed a series of template plasmids that carry repeated sequence of c-myc epitope, Flp recombinase target (FRT) sites, and apramycin resistance marker to insert epitope tags into any desired spot of the chromosomal loci. A DNA module which includes the tandem epitope-encoding sequence and a selectable marker was amplified by PCR with primers that carry homologous extensions to the last portion and downstream region of the targeted gene. We fused the epitope tags at the 3' region of global transcription factors of Streptomyces coelicolor to test the validity of this system. The proper insertion of the epitope tag was confirmed by PCR and western blot analysis. The recombinants showed the identical phenotype to the wild-type that proved the conservation of in vivo function of the tagged proteins. Finally, the direct binding targets were successfully detected by chromatin immunoprecipitation with the increase in the signal-to-noise ratio. The epitope tagging system describes here would provide wide applications to study the protein functions in S. coelicolor. PMID:22704935

  20. Positive Feedback Regulation of stgR Expression for Secondary Metabolism in Streptomyces coelicolor

    PubMed Central

    Mao, Xu-Ming; Sun, Zhi-Hao; Liang, Bi-Rong; Wang, Zhi-Bin; Feng, Wei-Hong; Huang, Fang-Liang

    2013-01-01

    LysR-type transcriptional regulators (LTTRs) compose a large family and are responsible for various physiological functions in bacteria, while little is understood about their regulatory mechanism on secondary metabolism in Streptomyces. Here we reported that StgR, a typical LTTR in Streptomyces coelicolor, was a negative regulator of undecylprodigiosin (Red) and γ-actinorhodin (Act) production in the early developmental phase of secondary metabolism by suppressing the expression of two pathway-specific regulator genes, redD and actII-orf4, respectively. Meanwhile, stgR expression was downregulated during secondary metabolism to remove its repressive effects on antibiotic production. Moreover, stgR expression was positively autoregulated by direct binding of StgR to its own promoter (stgRp), and the binding site adjacent to translation start codon was determined by a DNase I footprinting assay. Furthermore, the StgR-stgRp interaction could be destroyed by the antibiotic γ-actinorhodin produced from S. coelicolor. Thus, our results suggested a positive feedback regulatory mechanism of stgR expression and antibiotic production for the rapid and irreversible development of secondary metabolism in Streptomyces. PMID:23457252

  1. Cytochrome P450 107U1 is required for sporulation and antibiotic production in Streptomyces coelicolor

    PubMed Central

    Tian, Zhenghua; Cheng, Qian; Yoshimoto, Francis K.; Lei, Li; Lamb, David C.; Guengerich, F. Peter

    2013-01-01

    The filamentous bacterium Streptomyces coelicolor has a complex life cycle involving the formation of hair-like aerial mycelia on the colony surface, which differentiate into chains of spores. Genes required for the initiation of aerial mycelium formation have been termed ‘bld’ (bald), describing the smooth, undifferentiated colonies of mutant strains. We report the identification of a new bld gene designated as sco3099 and biochemical analysis of its encoded enzyme, cytochrome P450 (P450, or CYP) 107U1. Deletion of sco3099 resulted in a mutant defective in aerial hyphae sporulation and sensitive to heat shock, indicating that P450 107U1 plays a key role in growth and development of S. coelicolor. This is the first P450 reported to participate in a sporulation process in Streptomycetes. The substrate and catalytic properties of P450 107U1 were further investigated in mass spectrometry-based metabolomic studies. Glycocholic acid (from the medium) was identified as a substrate of P450 107U1 and was oxidized to glyco-7-oxo-deoxycholic acid. Although this reaction is apparently not relevant to the observed sporulation deficiency, it suggests that P450 107U1 might exert its physiological function by oxidizing other steroid-like molecules. PMID:23357279

  2. Developmentally regulated cleavage of tRNAs in the bacterium Streptomyces coelicolor

    PubMed Central

    Haiser, Henry J.; Karginov, Fedor V.; Hannon, Gregory J.; Elliot, Marie A.

    2008-01-01

    The ability to sense and respond to environmental and physiological signals is critical for the survival of the soil-dwelling Gram-positive bacterium Streptomyces coelicolor. Nutrient deprivation triggers the onset of a complex morphological differentiation process that involves the raising of aerial hyphae and formation of spore chains, and coincides with the production of a diverse array of clinically relevant antibiotics and other secondary metabolites. These processes are tightly regulated; however, the genes and signals involved have not been fully elucidated. Here, we report a novel tRNA cleavage event that follows the same temporal regulation as morphological and physiological differentiation, and is growth medium dependent. All tRNAs appear to be susceptible to cleavage; however, there appears to be a bias towards increased cleavage of those tRNAs that specify highly utilized codons. In contrast to what has been observed in eukaryotes, accumulation of tRNA halves in S. coelicolor is not significantly affected by amino acid starvation, and is also not affected by induction of the stringent response or inhibition of ribosome function. Mutants defective in aerial development and antibiotic production exhibit altered tRNA cleavage profiles relative to wild-type strains. PMID:18084030

  3. Expression of the melC Operon in Several Streptomyces Strains Is Positively Regulated by AdpA, an AraC Family Transcriptional Regulator Involved in Morphological Development in Streptomyces coelicolor

    PubMed Central

    Zhu, Dongqing; He, Xinyi; Zhou, Xiufen; Deng, Zixin

    2005-01-01

    Dark brown haloes of melanin around colonies are an easily visualized phenotype displayed by many Streptomyces strains harboring plasmid pIJ702 carrying the melC operon of Streptomyces antibioticus IMRU3270. Spontaneous melanin-negative mutants of pIJ702 occur with a frequency of ca. 1%, and often mutation occurs in the melC operon, which removes the BglII site as part of an inverted repeat. Other melanin-negative mutations seem to occur spontaneously in Streptomyces lividans, resulting in white colonies from which intact, melanin-producing pIJ702 can be isolated by introduction into a new host. S. lividans ZX66 was found to be such a mutant and to have a secondary mutation influencing expression of the melC operon on the chromosome. A 3.3-kb DNA fragment was isolated from its progenitor strain, JT46, and a gene able to restore melC operon expression was found to encode a member of an AraC family of transcriptional regulators, which was equivalent to AdpAc in Streptomyces coelicolor and therefore was designated AdpAl. Lack of melC operon expression was correlated with a single A-to-C transversion, which altered a single key amino acid residue from Thr to Pro. The transcription of the melC operon was found to be greatly reduced in the adpA mutant background. The counterpart gene (adpAa) in the S. antibioticus strain in which the melC operon carried on pIJ702 originated was also isolated and was found to have an identical regulatory role. Thus, we concluded that the melC operon is under general direct positive control by AdpA family proteins, perhaps at the transcriptional level and certainly at the translational level via bldA, in Streptomyces. PMID:15838045

  4. ArgR of Streptomyces coelicolor Is a Versatile Regulator

    PubMed Central

    Pérez-Redondo, Rosario; Rodríguez-García, Antonio; Botas, Alma; Santamarta, Irene; Martín, Juan F.; Liras, Paloma

    2012-01-01

    ArgR is the regulator of arginine biosynthesis genes in Streptomyces species. Transcriptomic comparison by microarrays has been made between Streptomyces coelicolor M145 and its mutant S. coelicolor ΔargR under control, unsupplemented conditions, and in the presence of arginine. Expression of 459 genes was different in transcriptomic assays, but only 27 genes were affected by arginine supplementation. Arginine and pyrimidine biosynthesis genes were derepressed by the lack of ArgR, while no strong effect on expression resulted on arginine supplementation. Several nitrogen metabolism genes expression as glnK, glnA and glnII, were downregulated in S. coelicolor ΔargR. In addition, downregulation of genes for the yellow type I polyketide CPK antibiotic and for the antibiotic regulatory genes afsS and scbR was observed. The transcriptomic data were validated by either reverse transcription-PCR, expression of the gene-promoter coupled to the luciferase gene, proteomic or by electrophoresis mobility shift assay (EMSA) using pure Strep-tagged ArgR. Two ARG-boxes in the arginine operon genes suggest that these genes are more tightly controlled. Other genes, including genes encoding regulatory proteins, possess a DNA sequence formed by a single ARG-box which responds to ArgR, as validated by EMSA. PMID:22403700

  5. Alternative-fueled truck demonstration natural gas program: Caterpillar G3406LE development and demonstration

    SciTech Connect

    1995-06-01

    In 1990, the California Energy Commission, the South Coast Air Quality Management District, and the Southern California Gas Company joined together to sponsor the development and demonstration of compressed natural gas engines for Class 8 heavy-duty line-haul trucking applications. This program became part of an overall Alternative-Fueled Truck Demonstration Program, with the goal of advancing the technological development of alternative-fueled engines. The demonstration showed natural gas to be a technically viable fuel for Class 8 truck engines.

  6. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 27 2011-07-01 2011-07-01 false Research, development, and... Special Forms of Permits § 270.65 Research, development, and demonstration permits. (a) The Administrator may issue a research, development, and demonstration permit for any hazardous waste treatment...

  7. 40 CFR 270.65 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 26 2010-07-01 2010-07-01 false Research, development, and... Special Forms of Permits § 270.65 Research, development, and demonstration permits. (a) The Administrator may issue a research, development, and demonstration permit for any hazardous waste treatment...

  8. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 4 2010-01-01 2010-01-01 false Research, development, demonstration, and testing. 1021..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  9. 17 CFR 256.188 - Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... demonstration expenditures. 256.188 Section 256.188 Commodity and Securities Exchanges SECURITIES AND EXCHANGE... demonstration expenditures. (a) The service company shall utilize this account, where appropriate, for the cost of all expenditures for research, development or demonstration undertaken by or sponsored through...

  10. Genetic and Proteomic Analyses of Pupylation in Streptomyces coelicolor

    PubMed Central

    Compton, Corey L.; Fernandopulle, Michael S.; Nagari, Rohith T.

    2015-01-01

    ABSTRACT Pupylation is a posttranslational modification peculiar to actinobacteria wherein proteins are covalently modified with a small protein called the prokaryotic ubiquitin-like protein (Pup). Like ubiquitination in eukaryotes, this phenomenon has been associated with proteasome-mediated protein degradation in mycobacteria. Here, we report studies of pupylation in a streptomycete that is phylogentically related to mycobacteria. We constructed mutants of Streptomyces coelicolor lacking PafA (Pup ligase), the proteasome, and the Pup-proteasome system. We found that these mutants share a high susceptibility to oxidative stress compared to that of the wild-type strain. Remarkably, we found that the pafA null mutant has a sporulation defect not seen in strains lacking the Pup-proteasome system. In proteomics experiments facilitated by an affinity-tagged variant of Pup, we identified 110 pupylated proteins in S. coelicolor strains having and lacking genes encoding the 20S proteasome. Our findings shed new light on this unusual posttranslational modification and its role in Streptomyces physiology. IMPORTANCE The presence of 20S proteasomes reminiscent of those in eukaryotes and a functional equivalent of ubiquitin, known as the prokaryotic ubiquitin-like protein (Pup), in actinobacteria have motivated reevaluations of protein homeostasis in prokaryotes. Though the Pup-proteasome system has been studied extensively in mycobacteria, it is much less understood in streptomycetes, members of a large genus of actinobacteria known for highly choreographed life cycles in which phases of morphological differentiation, sporulation, and secondary metabolism are often regulated by protein metabolism. Here, we define constituents of the pupylome in Streptomyces coelicolor for the first time and present new evidence that links pupylation and the oxidative stress response in this bacterium. Surprisingly, we found that the Pup ligase has a Pup-independent role in sporulation. PMID

  11. Structure of a 6-pyruvoyltetrahydropterin synthase homolog from Streptomyces coelicolor

    PubMed Central

    Spoonamore, James E.; Roberts, Sue A.; Heroux, Annie; Bandarian, Vahe

    2008-01-01

    The X-ray crystal structure of the 6-pyruvoyltetrahydropterin synthase (PTPS) homolog from Streptomyces coelicolor, SCO 6650, was solved at 1.5 Å resolution. SCO 6650 forms a hexameric T-fold that closely resembles other PTPS proteins. The biological activity of SCO 6650 is unknown, but it lacks both a required active-site zinc metal ion and the essential catalytic triad and does not catalyze the PTPS reaction. However, SCO 6650 maintains active-site residues consistent with binding a pterin-like substrate. PMID:18931427

  12. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 24 2010-07-01 2010-07-01 false Research, development, and demonstration permits. 258.4 Section 258.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and demonstration permits. (a) Except as...

  13. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 26 2013-07-01 2013-07-01 false Research, development, and demonstration permits. 258.4 Section 258.4 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and demonstration permits. (a) Except as...

  14. Structure and regulatory targets of SCO3201, a highly promiscuous TetR-like regulator of Streptomyces coelicolor M145.

    PubMed

    Xu, Delin; Waack, Paul; Zhang, Qizhong; Werten, Sebastiaan; Hinrichs, Winfried; Virolle, Marie-Joelle

    2014-07-18

    SCO3201, a regulator of the TetR family, is a strong repressor of both morphological differentiation and antibiotic production when overexpressed in Streptomyces coelicolor. Here, we report the identification of 14 novel putative regulatory targets of this regulator using in vitro formaldehyde cross-linking. Direct binding of purified His6-SCO3201 was demonstrated for the promoter regions of 5 regulators (SCO1716, SCO1950, SCO3367, SCO4009 and SCO5046), a putative histidine phosphatase (SCO1809), an acetyltransferase (SCO0988) and the polyketide synthase RedX (SCO5878), using EMSA. Reverse transcriptional analysis demonstrated that the expression of the transcriptional regulators SCO1950, SCO4009, SCO5046, as well as of SCO0988 and RedX was down regulated, upon SCO3201 overexpression, whereas the expression of SCO1809 and SCO3367 was up regulated. A consensus binding motif was derived via alignment of the promoter regions of the genes negatively regulated. The positions of the predicted operator sites were consistent with a direct repressive effect of SCO3201 on 5 out of 7 of these promoters. Furthermore, the 2.1Å crystal structure of SCO3201 was solved, which provides a possible explanation for the high promiscuity of this regulator that might account for its dramatic effect on the differentiation process of S. coelicolor. PMID:24928397

  15. New loci required for Streptomyces coelicolor morphological and physiological differentiation.

    PubMed Central

    Champness, W C

    1988-01-01

    Streptomyces coelicolor colonies differentiate both morphologically, producing aerial spore chains, and physiologically, producing antibiotics as secondary metabolites. Single mutations, which block both aspects of differentiation, define bld (bald colony) genes. To identify new bld genes, mutagenized colonies were screened for blocks in the earliest stage of sporulation, the formation of aerial mycelia, and blocks in antibiotic synthesis. The mutations in 12 mutants were mapped; in each strain, the pleiotropic phenotype was due to a single mutation. Seven of the strains contained mutations in known bld loci, bldA and bldB. Three strains contained mutations in a new locus, bldG, and two contained mutations in another new locus, bldH. Like the previously defined bldA mutants, the bldG and bldH mutants were developmentally blocked on glucose. On a variety of carbon sources whose utilization was subject to glucose repression, the developmental blocks were partially relieved for bldG (and bldA) mutants and fully relieved for bldH mutants. These results are compatible with an hypothesis which suggests that there are two alternative controls on S. coelicolor differentiation, one of which is glucose repressible. PMID:3343216

  16. 17 CFR 256.188 - Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 17 Commodity and Securities Exchanges 3 2011-04-01 2011-04-01 false Research, development, or... COMPANIES, PUBLIC UTILITY HOLDING COMPANY ACT OF 1935 4. Deferred Debits § 256.188 Research, development, or... of all expenditures for research, development or demonstration undertaken by or sponsored through...

  17. Codes and standards research, development and demonstration roadmap

    SciTech Connect

    None, None

    2008-07-22

    C&S RD&D Roadmap - 2008: This Roadmap is a guide to the Research, Development & Demonstration activities that will provide data required for Standards Development Organizations (SDOs) to develop performance-based codes and standards for a commercial hydrogen fueled transportation sector in the U.S.

  18. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 25 2011-07-01 2011-07-01 false Research, development, and...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... State may issue a research, development, and demonstration permit for a new MSWLF unit, existing...

  19. Experiments with Corn To Demonstrate Plant Growth and Development.

    ERIC Educational Resources Information Center

    Haldeman, Janice H.; Gray, Margarit S.

    2000-01-01

    Explores using corn seeds to demonstrate plant growth and development. This experiment allows students to formulate hypotheses, observe and record information, and practice mathematics. Presents background information, materials, procedures, and observations. (SAH)

  20. Deciphering the Regulon of Streptomyces coelicolor AbrC3, a Positive Response Regulator of Antibiotic Production

    PubMed Central

    Rico, Sergio; Santamaría, Ramón I.; Yepes, Ana; Rodríguez, Héctor; Laing, Emma; Bucca, Giselda; Smith, Colin P.

    2014-01-01

    The atypical two-component system (TCS) AbrC1/C2/C3 (encoded by SCO4598, SCO4597, and SCO4596), comprising two histidine kinases (HKs) and a response regulator (RR), is crucial for antibiotic production in Streptomyces coelicolor and for morphological differentiation under certain nutritional conditions. In this study, we demonstrate that deletion of the RR-encoding gene, abrC3 (SCO4596), results in a dramatic decrease in actinorhodin (ACT) and undecylprodiginine (RED) production and delays morphological development. In contrast, the overexpression of abrC3 in the parent strain leads to a 33% increase in ACT production in liquid medium. Transcriptomic analysis and chromatin immunoprecipitation with microarray technology (ChIP-chip) analysis of the ΔabrC3 mutant and the parent strain revealed that AbrC3 directly controls ACT production by binding to the actII-ORF4 promoter region; this was independently verified by in vitro DNA-binding assays. This binding is dependent on the sequence 5′-GAASGSGRMS-3′. In contrast, the regulation of RED production is not due to direct binding of AbrC3 to either the redZ or redD promoter region. This study also revealed other members of the AbrC3 regulon: AbrC3 is a positive autoregulator which also binds to the promoter regions of SCO0736, bdtA (SCO3328), absR1 (SCO6992), and SCO6809. The direct targets share the 10-base consensus binding sequence and may be responsible for some of the phenotypes of the ΔabrC3 mutant. The identification of the AbrC3 regulon as part of the complex regulatory network governing antibiotic production widens our knowledge regarding TCS involvement in control of antibiotic synthesis and may contribute to the rational design of new hyperproducer host strains through genetic manipulation of such systems. PMID:24509929

  1. Structural Insight into How Streptomyces coelicolor Maltosyl Transferase GlgE Binds α-Maltose 1-Phosphate and Forms a Maltosyl-enzyme Intermediate

    PubMed Central

    2014-01-01

    GlgE (EC 2.4.99.16) is an α-maltose 1-phosphate:(1→4)-α-d-glucan 4-α-d-maltosyltransferase of the CAZy glycoside hydrolase 13_3 family. It is the defining enzyme of a bacterial α-glucan biosynthetic pathway and is a genetically validated anti-tuberculosis target. It catalyzes the α-retaining transfer of maltosyl units from α-maltose 1-phosphate to maltooligosaccharides and is predicted to use a double-displacement mechanism. Evidence of this mechanism was obtained using a combination of site-directed mutagenesis of Streptomyces coelicolor GlgE isoform I, substrate analogues, protein crystallography, and mass spectrometry. The X-ray structures of α-maltose 1-phosphate bound to a D394A mutein and a β-2-deoxy-2-fluoromaltosyl-enzyme intermediate with a E423A mutein were determined. There are few examples of CAZy glycoside hydrolase family 13 members that have had their glycosyl-enzyme intermediate structures determined, and none before now have been obtained with a 2-deoxy-2-fluoro substrate analogue. The covalent modification of Asp394 was confirmed using mass spectrometry. A similar modification of wild-type GlgE proteins from S. coelicolor and Mycobacterium tuberculosis was also observed. Small-angle X-ray scattering of the M. tuberculosis enzyme revealed a homodimeric assembly similar to that of the S. coelicolor enzyme but with slightly differently oriented monomers. The deeper understanding of the structure–function relationships of S. coelicolor GlgE will aid the development of inhibitors of the M. tuberculosis enzyme. PMID:24689960

  2. Development Status of the CECE Cryogenic Deep Throttling Demonstrator Engine

    NASA Technical Reports Server (NTRS)

    2008-01-01

    As one of the first technology development programs awarded by NASA under the U.S. Space Exploration Policy (USSEP), the Pratt & Whitney Rocketdyne (PWR) Deep Throttling, Common Extensible Cryogenic Engine (CECE) program was selected by NASA in November 2004 to begin technology development and demonstration toward a deep throttling, cryogenic engine supporting ongoing trade studies for NASA's Lunar Lander descent stage. The CECE program leverages the maturity and previous investment of a flight-proven hydrogen/oxygen expander cycle engine, the PWR RLI0, to develop and demonstrate an unprecedented combination of reliability, safety, durability, throttlability, and restart capabilities in a high-energy, cryogenic engine. The testbed selected for the deep throttling demonstration phases of this program was a minimally modified RL10 engine, allowing for maximum current production engine commonality and extensibility with minimum program cost. Two series of demonstrator engine tests, the first in April-May 2006 and the second in March-April 2007, have demonstrated in excess of 10:1 throttling of the hydrogen/oxygen expander cycle engine. Both test series have explored a combustion instability ("chug") environment at low throttled power levels. These tests have provided an early demonstration of an enabling cryogenic propulsion concept with invaluable system-level technology data acquisition toward design and development risk mitigation for future CECE Demonstrator engine tests.

  3. The Phosphotransferase System of Streptomyces coelicolor Is Biased for N-Acetylglucosamine Metabolism

    PubMed Central

    Nothaft, Harald; Dresel, Dagmar; Willimek, Andreas; Mahr, Kerstin; Niederweis, Michael; Titgemeyer, Fritz

    2003-01-01

    Mutation of the crr-ptsI gene locus revealed that Streptomyces coelicolor uses the phosphotransferase system (PTS) for N-acetylglucosamine uptake. crr, ptsI, and ptsH, which encode the three general PTS phosphotransferases, are induced by N-acetylglucosamine but not by other PTS substrates. Thus, the S. coelicolor PTS is biased for N-acetylglucosamine utilization, a novel feature that distinguishes this PTS from others. PMID:14617669

  4. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs might also lead to commercialization or other broad-scale implementation by DOE or another entity. (b... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  5. 10 CFR 1021.212 - Research, development, demonstration, and testing.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ..., development, demonstration, and testing for new technologies (40 CFR 1502.4(c)(3)). Adoption of such programs might also lead to commercialization or other broad-scale implementation by DOE or another entity. (b... development or restrict later alternatives, as discussed at 40 CFR 1502.4(c)(3). (c) For subsequent phases...

  6. New Sporulation Loci in Streptomyces coelicolor A3(2)

    PubMed Central

    Ryding, N. Jamie; Bibb, Maureen J.; Molle, Virginie; Findlay, Kim C.; Chater, Keith F.; Buttner, Mark J.

    1999-01-01

    Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the grey polyketide spore pigment, and such white (whi) mutants had been used to define eight sporulation loci, whiA, whiB, whiD, whiE, whiG, whiH, whiI, and whiJ (K. F. Chater, J. Gen. Microbiol. 72:9–28, 1972; N. J. Ryding, Ph.D. thesis, University of East Anglia, 1995). In an attempt to identify new whi loci, we mutagenized S. coelicolor M145 spores with nitrosoguanidine and identified 770 mutants with colonies ranging from white to medium grey. After excluding unstable strains, we examined the isolates by phase-contrast microscopy and chose 115 whi mutants with clear morphological phenotypes for further study. To exclude mutants representing cloned whi genes, self-transmissible SCP2*-derived plasmids carrying whiA, whiB, whiG, whiH, or whiJ (but not whiD, whiE, or whiI) were introduced into each mutant by conjugation, and strains in which the wild-type phenotype was restored either partially or completely by any of these plasmids were excluded from further analysis. In an attempt to complement some of the remaining 31 whi mutants, an SCP2* library of wild-type S. coelicolor chromosomal DNA was introduced into 19 of the mutants by conjugation. Clones restoring the wild-type phenotype to 12 of the 19 strains were isolated and found to represent five distinct loci, designated whiK, whiL, whiM, whiN, and whiO. Each of the five loci was located on the ordered cosmid library: whiL, whiM, whiN, and whiO occupied positions distinct from previously cloned whi genes; whiK was located on the same cosmid overlap as whiD, but the two loci were shown by complementation to be distinct. The phenotypes resulting from mutations at each of these new loci are described. PMID:10464216

  7. Integrated gasification combined-cycle research development and demonstration activities

    SciTech Connect

    Ness, H.M.; Reuther, R.B.

    1995-12-01

    The United States Department of Energy (DOE) has selected six integrated gasification combined-cycle (IGCC) advanced power systems for demonstration in the Clean Coal Technology (CCT) Program. DOE`s Office of Fossil Energy, Morgantown Energy Technology Center, is managing a research development and demonstration (RD&D) program that supports the CCT program, and addresses long-term improvements in support of IGCC technology. This overview briefly describes the CCT projects and the supporting RD&D activities.

  8. Cloning, disruption, and transcriptional analysis of three RNA polymerase sigma factor genes of Streptomyces coelicolor A3(2).

    PubMed Central

    Buttner, M J; Chater, K F; Bibb, M J

    1990-01-01

    The rpoD gene of Myxococcus xanthus was used as a probe to isolate three Streptomyces coelicolor genes, hrdB, hrdC, and hrdD, which appear to encode RNA polymerase sigma factors extremely similar to the sigma 70 polypeptide of Escherichia coli. Gene disruption experiments suggested that hrdB is essential in S. coelicolor A3(2) but showed that hrdC and hrdD mutants are viable and are apparently unaffected in differentiation, gross morphology, and antibiotic production. S1 nuclease mapping showed that hrdB and hrdD, but not hrdC, were transcribed in liquid culture. The most upstream of two hrdD promoters is internal to an open reading frame (ORF X) on the opposite strand. The predicted product of this gene is homologous to the phosphinothricin acetyltransferases of Streptomyces hygroscopicus and Streptomyces viridochromogenes. The possible significance of the overlapping and divergent transcription of hrdD and ORF X is discussed. A general method for in vivo gene replacement was developed that allowed a positive selection for the desired mutants even in the absence of a mutant phenotype; it was used to isolate a stable hrdC mutant. Images PMID:2160942

  9. Satellite Communications for Aeronautics Applications: Technology Development and Demonstration

    NASA Technical Reports Server (NTRS)

    Kerczewski, Robert J.; Hoder, Douglas J.; Zakrajsek, Robert J.

    2001-01-01

    The National Aeronautics and Space Administration (NASA) is performing research and development to improve the safety and increase the capacity of the National Airspace System (NAS). Improved communications, especially to and from the aircraft flight deck, has been identified as an essential enabling technology for future improvements to the air traffic management system and aviation safety. NASA's Glenn Research Center is engaged in research and development of satellite communications technologies for aeronautical applications. A mobile aero terminal has been developed for use with Ku band commercial communications satellites. This experimental terminal will be used in mobile ground and air-based tests and demonstrations during 2000-2004. This paper will describe the basic operational parameters of the Ku Band aero terminal, the communications architecture it is intended to demonstrate, and the key technology issues being addressed in the tests and demonstrations. The design of the Ku Band aero terminal and associated ground testbed, planned tests and demonstrations, and results to date will be presented.

  10. Aging Aircraft NDI Development and Demonstration Center (AANC): An overview

    SciTech Connect

    Walter, P.L.

    1991-01-01

    A major center with emphasis on validation of nondestructive inspection techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing test beds for nondestructive inspection validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed. 3 refs.

  11. Aging aircraft NDI Development and Demonstration Center (AANC): An overview

    NASA Astrophysics Data System (ADS)

    Walter, Patrick L.

    1992-07-01

    A major center with emphasis on validation of nondestructive inspection (NDI) techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing testbeds for NDI validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed.

  12. Advanced Simulation Capability for Environmental Management: Development and Demonstrations - 12532

    SciTech Connect

    Freshley, Mark D.; Freedman, Vicky; Gorton, Ian; Hubbard, Susan S.; Moulton, J. David; Dixon, Paul

    2012-07-01

    The U.S. Department of Energy Office of Environmental Management (EM), Technology Innovation and Development is supporting development of the Advanced Simulation Capability for Environmental Management (ASCEM). ASCEM is a state-of-the-art scientific tool and approach for understanding and predicting contaminant fate and transport in natural and engineered systems. The modular and open source high-performance computing tool facilitates integrated approaches to modeling and site characterization that enable robust and standardized assessments of performance and risk for EM cleanup and closure activities. The ASCEM project continues to make significant progress in development of capabilities, which are organized into Platform and Integrated Tool-sets and a High-Performance Computing Multi-process Simulator. The Platform capabilities target a level of functionality to allow end-to-end model development, starting with definition of the conceptual model and management of data for model input. The High-Performance Computing capabilities target increased functionality of process model representations, tool-sets for interaction with Platform, and verification and model confidence testing. The new capabilities are demonstrated through working groups, including one focused on the Hanford Site Deep Vadose Zone. The ASCEM program focused on planning during the first year and executing a prototype tool-set for an early demonstration of individual components. Subsequently, ASCEM has focused on developing and demonstrating an integrated set of capabilities, making progress toward a version of the capabilities that can be used to engage end users. Demonstration of capabilities continues to be implemented through working groups. Three different working groups, one focused on EM problems in the deep vadose zone, another investigating attenuation mechanisms for metals and radionuclides, and a third focusing on waste tank performance assessment, continue to make progress. The project

  13. Technology Development and Demonstration Concepts for the Space Elevator

    NASA Technical Reports Server (NTRS)

    Smitherman, David V., Jr.

    2004-01-01

    During the 1990s several discoveries and advances in the development of carbon nano-tube (CNT) materials indicated that material strengths many times greater than common high-strength composite materials might be possible. Progress in the development of this material led to renewed interest in the space elevator concept for construction of a tether structure from the surface of the Earth through a geostationary orbit (GEO) and thus creating a new approach to Earth-to-orbit transportation infrastructures. To investigate this possibility the author, in 1999, managed for NASA a space elevator work:hop at the Marshall Space Flight Center to explore the potential feasibility of space elevators in the 21 century, and to identify the critical technologies and demonstration missions needed to make development of space elevators feasible. Since that time, a NASA Institute for Advanced Concepts (NIAC) funded study of the Space Elevator proposed a concept for a simpler first space elevator system using more near-term technologies. This paper will review some of the latest ideas for space elevator development, the critical technologies required, and some of the ideas proposed for demonstrating the feasibility for full-scale development of an Earth to GEO space elevator. Critical technologies include CNT composite materials, wireless power transmission, orbital object avoidance, and large-scale tether deployment and control systems. Numerous paths for technology demonstrations have been proposed utilizing ground experiments, air structures. LEO missions, the space shuttle, the international Space Station, GEO demonstration missions, demonstrations at the lunar L1 or L2 points, and other locations. In conclusion, this paper finds that the most critical technologies for an Earth to GEO space elevator include CNT composite materials development and object avoidance technologies; that lack of successful development of these technologies need not preclude continued development of

  14. INTERNATIONAL ENVIRONMENTAL TECHNOLOGY IDENTIFICATION, DEVELOPMENT, DEMONSTRATION, DEPLOYMENT AND EXCHANGE

    SciTech Connect

    Roy C. Herndon

    2001-02-28

    Cooperative Agreement (DE-FC21-95EW55101) between the U.S. Department of Energy (DOE) and the Florida State University's Institute for International Cooperative Environmental Research (IICER) was designed to facilitate a number of joint programmatic goals of both the DOE and the IICER related to international technology identification, development, demonstration and deployment using a variety of mechanisms to accomplish these goals. These mechanisms included: laboratory and field research; technology demonstrations; international training and technical exchanges; data collection, synthesis and evaluation; the conduct of conferences, symposia and high-level meetings; and other appropriate and effective approaches. The DOE utilized the expertise and facilities of the IICER at Florida State University to accomplish its goals related to this cooperative agreement. The IICER has unique and demonstrated capabilities that have been utilized to conduct the tasks for this cooperative agreement. The IICER conducted activities related to technology identification, development, evaluation, demonstration and deployment through its joint centers which link the capabilities at Florida State University with collaborating academic and leading research institutions in the major countries of Central and Eastern Europe (e.g., Czech Republic, Hungary, Poland) and Russia. The activities and accomplishments for this five-year cooperative agreement are summarized in this Final Technical Report.

  15. Texas LPG fuel cell development and demonstration project

    SciTech Connect

    None, None

    2004-07-26

    The State Energy Conservation Office has executed its first Fuel Cell Project which was awarded under a Department of Energy competitive grant process. The Texas LPG Fuel Processor Development and Fuel Cell Demonstration Program is a broad-based public/private partnership led by the Texas State Energy Conservation Office (SECO). Partners include the Alternative Fuels Research and Education Division (AFRED) of the Railroad Commission of Texas; Plug Power, Inc., Latham, NY, UOP/HyRadix, Des Plaines, IL; Southwest Research Institute (SwRI), San Antonio, TX; the Texas Natural Resource Conservation Commission (TNRCC), and the Texas Department of Transportation (TxDOT). The team proposes to mount a development and demonstration program to field-test and evaluate markets for HyRadix's LPG fuel processor system integrated into Plug Power's residential-scale GenSys(TM) 5C (5 kW) PEM fuel cell system in a variety of building types and conditions of service. The program's primary goal is to develop, test, and install a prototype propane-fueled residential fuel cell power system supplied by Plug Power and HyRadix in Texas. The propane industry is currently funding development of an optimized propane fuel processor by project partner UOP/HyRadix through its national checkoff program, the Propane Education and Research Council (PERC). Following integration and independent verification of performance by Southwest Research Institute, Plug Power and HyRadix will produce a production-ready prototype unit for use in a field demonstration. The demonstration unit produced during this task will be delivered and installed at the Texas Department of Transportation's TransGuide headquarters in San Antonio, Texas. Simultaneously, the team will undertake a market study aimed at identifying and quantifying early-entry customers, technical and regulatory requirements, and other challenges and opportunities that need to be addressed in planning commercialization of the units. For further

  16. Clean coal technologies: Research, development, and demonstration program plan

    SciTech Connect

    Not Available

    1993-12-01

    The US Department of Energy, Office of Fossil Energy, has structured an integrated program for research, development, and demonstration of clean coal technologies that will enable the nation to use its plentiful domestic coal resources while meeting environmental quality requirements. The program provides the basis for making coal a low-cost, environmentally sound energy choice for electric power generation and fuels production. These programs are briefly described.

  17. Demonstration of Decision Support Tools for Sustainable Development

    SciTech Connect

    Shropshire, David Earl; Jacobson, Jacob Jordan; Berrett, Sharon; Cobb, D. A.; Worhach, P.

    2000-11-01

    The Demonstration of Decision Support Tools for Sustainable Development project integrated the Bechtel/Nexant Industrial Materials Exchange Planner and the Idaho National Engineering and Environmental Laboratory System Dynamic models, demonstrating their capabilities on alternative fuel applications in the Greater Yellowstone-Teton Park system. The combined model, called the Dynamic Industrial Material Exchange, was used on selected test cases in the Greater Yellow Teton Parks region to evaluate economic, environmental, and social implications of alternative fuel applications, and identifying primary and secondary industries. The test cases included looking at compressed natural gas applications in Teton National Park and Jackson, Wyoming, and studying ethanol use in Yellowstone National Park and gateway cities in Montana. With further development, the system could be used to assist decision-makers (local government, planners, vehicle purchasers, and fuel suppliers) in selecting alternative fuels, vehicles, and developing AF infrastructures. The system could become a regional AF market assessment tool that could help decision-makers understand the behavior of the AF market and conditions in which the market would grow. Based on this high level market assessment, investors and decision-makers would become more knowledgeable of the AF market opportunity before developing detailed plans and preparing financial analysis.

  18. Development and demonstration of autonomous behaviors for urban environment exploration

    NASA Astrophysics Data System (ADS)

    Ahuja, Gaurav; Fellars, Donald; Kogut, Gregory; Pacis Rius, Estrellina; Schoolov, Misha; Xydes, Alexander

    2012-06-01

    Under the Urban Environment Exploration project, the Space and Naval Warfare Systems Center Pacic (SSC- PAC) is maturing technologies and sensor payloads that enable man-portable robots to operate autonomously within the challenging conditions of urban environments. Previously, SSC-PAC has demonstrated robotic capabilities to navigate and localize without GPS and map the ground oors of various building sizes.1 SSC-PAC has since extended those capabilities to localize and map multiple multi-story buildings within a specied area. To facilitate these capabilities, SSC-PAC developed technologies that enable the robot to detect stairs/stairwells, maintain localization across multiple environments (e.g. in a 3D world, on stairs, with/without GPS), visualize data in 3D, plan paths between any two points within the specied area, and avoid 3D obstacles. These technologies have been developed as independent behaviors under the Autonomous Capabilities Suite, a behavior architecture, and demonstrated at a MOUT site at Camp Pendleton. This paper describes the perceptions and behaviors used to produce these capabilities, as well as an example demonstration scenario.

  19. Development and demonstration of the TRUEX solvent extraction process

    SciTech Connect

    Vandegrift, G.F.; Chamberlain, D.B.; Conner, C.; Copple, J.M.; Dow, J.A.; Everson, L.; Hutter, J.C.; Leonard, R.A.; Nunez, L.; Regalbuto, M.C.; Sedlet, J.; Srinivasan, B.; Weber, S.; Wygmans, D.G.

    1993-03-01

    The Generic TRUEX Model (GTM) was developed for use in designing site and feed-specific TRUEX flowsheets and in estimating the space and cost requirements for installing a TRUEX process. This paper discusses data collected in support of the GTM and its use in (1) designing process flowsheets and (2) performing sensitivity analyses. Demonstration of the TRUEX process is underway at Argonne National Laboratory (ANL), where plutonium-containing analytical waste solutions generated at the DOE New Brunswick Laboratory (NBL) are being converted from TRU waste ({approximately}l g Pu/L), with no current means of disposal, to solutions that contain less than 10 nCi of transuranic elements per milliliter of waste solution. Results and implications of this demonstration are discussed in this paper.

  20. Development and demonstration of the TRUEX solvent extraction process

    SciTech Connect

    Vandegrift, G.F.; Chamberlain, D.B.; Conner, C.; Copple, J.M.; Dow, J.A.; Everson, L.; Hutter, J.C.; Leonard, R.A.; Nunez, L.; Regalbuto, M.C.; Sedlet, J.; Srinivasan, B.; Weber, S.; Wygmans, D.G.

    1993-01-01

    The Generic TRUEX Model (GTM) was developed for use in designing site and feed-specific TRUEX flowsheets and in estimating the space and cost requirements for installing a TRUEX process. This paper discusses data collected in support of the GTM and its use in (1) designing process flowsheets and (2) performing sensitivity analyses. Demonstration of the TRUEX process is underway at Argonne National Laboratory (ANL), where plutonium-containing analytical waste solutions generated at the DOE New Brunswick Laboratory (NBL) are being converted from TRU waste ([approximately]l g Pu/L), with no current means of disposal, to solutions that contain less than 10 nCi of transuranic elements per milliliter of waste solution. Results and implications of this demonstration are discussed in this paper.

  1. The development and demonstration of hybrid programmable attitude control electronics

    NASA Technical Reports Server (NTRS)

    Smith, L. S.; Kopf, E. H., Jr.

    1973-01-01

    In the course of extended life attitude control system (ELACS) research sponsored by NASA a hybrid programable attitude control electronics (HYPACE) concept was developed and demonstrated. The wide variety of future planetary missions demanded a new control approach to accommodate the automatic fault tolerance and long the life requirements of such missions. HYPACE provides an adaptable, analog/digital design approach that permits preflight and in-flight accommodation of mission changes, component performance variations, and spacecraft changes, through programing. This enabled broad multimission flexibility of application in a cost effective manner. Previously, flight control computers have not been not flown on planetary missions because of weight and power problems. These problems were resolved in the design of HYPACE. The HYPACE design, which was demonstrated in breadboard form on a single-axis gas-bearing spacecraft simulation, uses a single control channel to perform the attitude control functions sequentially, thus significantly reducing the number of component parts over hard-wired designs.

  2. Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol-xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum.

    PubMed

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-03-01

    Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol-xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies. PMID:23163932

  3. Secretory production of an FAD cofactor-containing cytosolic enzyme (sorbitol–xylitol oxidase from Streptomyces coelicolor) using the twin-arginine translocation (Tat) pathway of Corynebacterium glutamicum

    PubMed Central

    Scheele, Sandra; Oertel, Dan; Bongaerts, Johannes; Evers, Stefan; Hellmuth, Hendrik; Maurer, Karl-Heinz; Bott, Michael; Freudl, Roland

    2013-01-01

    Carbohydrate oxidases are biotechnologically interesting enzymes that require a tightly or covalently bound cofactor for activity. Using the industrial workhorse Corynebacterium glutamicum as the expression host, successful secretion of a normally cytosolic FAD cofactor-containing sorbitol–xylitol oxidase from Streptomyces coelicolor was achieved by using the twin-arginine translocation (Tat) protein export machinery for protein translocation across the cytoplasmic membrane. Our results demonstrate for the first time that, also for cofactor-containing proteins, a secretory production strategy is a feasible and promising alternative to conventional intracellular expression strategies. PMID:23163932

  4. The dynamic transcriptional and translational landscape of the model antibiotic producer Streptomyces coelicolor A3(2)

    PubMed Central

    Jeong, Yujin; Kim, Ji-Nu; Kim, Min Woo; Bucca, Giselda; Cho, Suhyung; Yoon, Yeo Joon; Kim, Byung-Gee; Roe, Jung-Hye; Kim, Sun Chang; Smith, Colin P.; Cho, Byung-Kwan

    2016-01-01

    Individual Streptomyces species have the genetic potential to produce a diverse array of natural products of commercial, medical and veterinary interest. However, these products are often not detectable under laboratory culture conditions. To harness their full biosynthetic potential, it is important to develop a detailed understanding of the regulatory networks that orchestrate their metabolism. Here we integrate nucleotide resolution genome-scale measurements of the transcriptome and translatome of Streptomyces coelicolor, the model antibiotic-producing actinomycete. Our systematic study determines 3,570 transcription start sites and identifies 230 small RNAs and a considerable proportion (∼21%) of leaderless mRNAs; this enables deduction of genome-wide promoter architecture. Ribosome profiling reveals that the translation efficiency of secondary metabolic genes is negatively correlated with transcription and that several key antibiotic regulatory genes are translationally induced at transition growth phase. These findings might facilitate the design of new approaches to antibiotic discovery and development. PMID:27251447

  5. The dynamic transcriptional and translational landscape of the model antibiotic producer Streptomyces coelicolor A3(2).

    PubMed

    Jeong, Yujin; Kim, Ji-Nu; Kim, Min Woo; Bucca, Giselda; Cho, Suhyung; Yoon, Yeo Joon; Kim, Byung-Gee; Roe, Jung-Hye; Kim, Sun Chang; Smith, Colin P; Cho, Byung-Kwan

    2016-01-01

    Individual Streptomyces species have the genetic potential to produce a diverse array of natural products of commercial, medical and veterinary interest. However, these products are often not detectable under laboratory culture conditions. To harness their full biosynthetic potential, it is important to develop a detailed understanding of the regulatory networks that orchestrate their metabolism. Here we integrate nucleotide resolution genome-scale measurements of the transcriptome and translatome of Streptomyces coelicolor, the model antibiotic-producing actinomycete. Our systematic study determines 3,570 transcription start sites and identifies 230 small RNAs and a considerable proportion (∼21%) of leaderless mRNAs; this enables deduction of genome-wide promoter architecture. Ribosome profiling reveals that the translation efficiency of secondary metabolic genes is negatively correlated with transcription and that several key antibiotic regulatory genes are translationally induced at transition growth phase. These findings might facilitate the design of new approaches to antibiotic discovery and development. PMID:27251447

  6. Microbial Transformation of Antibiotics: Phosphorylation of Clindamycin by Streptomyces coelicolor Müller1

    PubMed Central

    Coats, John H.; Argoudelis, Alexander D.

    1971-01-01

    Addition of clindamycin to whole-cell cultures of Streptomyces coelicolor Müller resulted in the loss of in vitro activity against organisms sensitive to clindamycin. Incubation of such culture filtrates with alkaline phosphatase generated a biologically active material identified as clindamycin. Fermentation broths containing inactivated clindamycin yielded clindamycin 3-phosphate, the structure of which was established by physical-chemical and enzymatic studies. Clindamycin was phosphorylated by lysates and partially purified enzyme preparations from S. coelicolor Müller. These reactions require a ribonucleoside triphosphate and Mg2+. The product of the cell-free reactions was identified as clindamycin 3-phosphate. PMID:5166238

  7. Effect of PCL/PEG-Based Membranes on Actinorhodin Production in Streptomyces coelicolor Cultivations.

    PubMed

    Scaffaro, Roberto; Lopresti, Francesco; Sutera, Alberto; Botta, Luigi; Fontana, Rosa Maria; Puglia, Anna Maria; Gallo, Giuseppe

    2016-05-01

    The actinomycetes, Gram-positive filamentous bacteria, are the most prolific source of natural occurring antibiotics. At an industrial level, antibiotics from actinomycete strains are produced by means of submerged fermentations, where one of the major factors negatively affecting bioproductivity is the pellet-shaped biomass growth. The immobilization of microorganisms on properly chosen supports prevents cell-cell aggregation resulting in improving the biosynthetic capability. Thus, novel porous biopolymer-based devices are developed by combining melt mixing and particulate leaching. In particular, polycaprolactone (PCL), polyethylene glycol (PEG), and sodium chloride (NaCl) with different grain sizes are used to prepare PCL/PEG/NaCl blends in the melt. These blends are then leached to obtain PCL-based porous membranes that are used as solid support for the growth of Streptomyces coelicolor, a model streptomycete used to produce various antibiotics including the blue colored actinorhodin (ACT). Thereafter, the effect of the devices' characteristics on the bacterial growth and on the production ACT is evaluated. The results showed that ACT production is strongly dependent on the pore size distribution of the device. Moreover, membranes with pores ranging from 90 to 110 μm are able to offer a potential improvement in volumetric productivity of ACT if compared to conventional submerged liquid culture. PMID:26762618

  8. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2).

    PubMed

    Bentley, S D; Chater, K F; Cerdeño-Tárraga, A-M; Challis, G L; Thomson, N R; James, K D; Harris, D E; Quail, M A; Kieser, H; Harper, D; Bateman, A; Brown, S; Chandra, G; Chen, C W; Collins, M; Cronin, A; Fraser, A; Goble, A; Hidalgo, J; Hornsby, T; Howarth, S; Huang, C-H; Kieser, T; Larke, L; Murphy, L; Oliver, K; O'Neil, S; Rabbinowitsch, E; Rajandream, M-A; Rutherford, K; Rutter, S; Seeger, K; Saunders, D; Sharp, S; Squares, R; Squares, S; Taylor, K; Warren, T; Wietzorrek, A; Woodward, J; Barrell, B G; Parkhill, J; Hopwood, D A

    2002-05-01

    Streptomyces coelicolor is a representative of the group of soil-dwelling, filamentous bacteria responsible for producing most natural antibiotics used in human and veterinary medicine. Here we report the 8,667,507 base pair linear chromosome of this organism, containing the largest number of genes so far discovered in a bacterium. The 7,825 predicted genes include more than 20 clusters coding for known or predicted secondary metabolites. The genome contains an unprecedented proportion of regulatory genes, predominantly those likely to be involved in responses to external stimuli and stresses, and many duplicated gene sets that may represent 'tissue-specific' isoforms operating in different phases of colonial development, a unique situation for a bacterium. An ancient synteny was revealed between the central 'core' of the chromosome and the whole chromosome of pathogens Mycobacterium tuberculosis and Corynebacterium diphtheriae. The genome sequence will greatly increase our understanding of microbial life in the soil as well as aiding the generation of new drug candidates by genetic engineering. PMID:12000953

  9. Role of Acid Metabolism in Streptomyces coelicolor Morphological Differentiation and Antibiotic Biosynthesis

    PubMed Central

    Viollier, Patrick H.; Minas, Wolfgang; Dale, Glenn E.; Folcher, Marc; Thompson, Charles J.

    2001-01-01

    Studies of citrate synthase (CitA) were carried out to investigate its role in morphological development and biosynthesis of antibiotics in Streptomyces coelicolor. Purification of CitA, the major vegetative enzyme activity, allowed characterization of its kinetic properties. The apparent Km values of CitA for acetyl coenzyme A (acetyl-CoA) (32 μM) and oxaloacetate (17 μM) were similar to those of citrate synthases from other gram-positive bacteria and eukaryotes. CitA was not strongly inhibited by various allosteric feedback inhibitors (NAD+, NADH, ATP, ADP, isocitrate, or α-ketoglutarate). The corresponding gene (citA) was cloned and sequenced, allowing construction of a citA mutant (BZ2). BZ2 was a glutamate auxotroph, indicating that citA encoded the major citrate synthase allowing flow of acetyl-CoA into the tricarboxylic acid (TCA) cycle. Interruption of aerobic TCA cycle-based metabolism resulted in acidification of the medium and defects in morphological differentiation and antibiotic biosynthesis. These developmental defects of the citA mutant were in part due to a glucose-dependent medium acidification that was also exhibited by some other bald mutants. Unlike other acidogenic bald strains, citA and bldJ mutants were able to produce aerial mycelia and pigments when the medium was buffered sufficiently to maintain neutrality. Extracellular complementation studies suggested that citA defines a new stage of the Streptomyces developmental cascade. PMID:11325948

  10. A jumping gene in streptomyces coelicolor A3(2).

    PubMed

    Sermonti, G; Lanfaloni, L; Micheli, M R

    1980-02-01

    The difficulty in mapping the gene for chloramphenicol resistance (cmlR) in Streptomyces coelicolor A3(2) stock strains is possibly due to its location on different sites of the chromosome in various mixed subclones. Fresh isolates from CmlR strains show single unequivocal locations of cmlR. The same holds for CmlR strains derived as revertants from CmlS variants. The two best established sites for cmlR are one between cysA and metA, the other at right of argA, possibly in the right empty arc of the map (Fig. 2). The cmlR gene was assumed to be on a transposon (SCTn1), together with a gene for arginine-succinate synthase (argG), a gene for chromosome transfer (tra) and a gene for aereal mycelium formation (amy). In a CmlR revertant, the cmlR gene appears disjoined from argG (Fig. 5), thus showing the ability of SCTnl to be split and partially transposed. The possible wide occurrence of transposons in the genus Streptomyces is discussed. PMID:6246399

  11. Development and demonstration of a telerobotic excavation system

    NASA Technical Reports Server (NTRS)

    Burks, Barry L.; Thompson, David H.; Killough, Stephen M.; Dinkins, Marion A.

    1994-01-01

    Oak Ridge National Laboratory is developing remote excavation technologies for the Department of Energy's Office (DOE) of Technology Development, Robotics Technology Development Program, and also for the Department of Defense (DOD) Project Manager for Ammunition Logistics. This work is being done to meet the need for remote excavation and removal of radioactive and contaminated buried waste at several DOE sites and unexploded ordnance at DOD sites. System requirements are based on the need to uncover and remove waste from burial sites in a way that does not cause unnecessary personnel exposure or additional environmental contamination. Goals for the current project are to demonstrate dexterous control of a backhoe with force feedback and to implement robotic operations that will improve productivity. The Telerobotic Small Emplacement Excavator is a prototype system that incorporates the needed robotic and telerobotic capabilities on a commercially available platform. The ability to add remote dexterous teleoperation and robotic operating modes is intended to be adaptable to other commercially available excavator systems.

  12. Technology development and demonstration of a low thrust resistojet thruster

    NASA Technical Reports Server (NTRS)

    Pfeifer, G. R.

    1972-01-01

    Three thrusters were fabricated to definitized thruster drawings using new rhenium vapor deposition technology. Two of the thrusters were operated using ammonia as propellant and one was operated using hydrogen propellant for performance determination. All demonstrated consistent operational specific impulse performance while demonstrating thermal performance better than the development units from which they evolved. Two of the thrusters were subjected to environmental structural testing including vibration, acceleration and shock loading to specifications. Both of the thrusters subjected to the environmental tests passed all required tests. The third, spare, thruster was introduced into the life test portion of the program. Two thrusters were then subjected to a life cycling test program under typical spacecraft operating power levels. During the life test sequence, the hydrogen thruster accrued 720 operating life test cycles, more than 370 on-off cycles and 365 hours of powered up time. The ammonia accrued approximately 380 on-off cycles and 392.2 on time hours of operation during the 720 cycling hour test. Both thrusters completed the scheduled operational life test in reasonably good condition, structurally integral and capable of indefinite further operation.

  13. Development Status of the Fission Power System Technology Demonstration Unit

    NASA Technical Reports Server (NTRS)

    Briggs, Maxwell H.; Gibson, Marc A.; Geng, Steven M.; Pearson, Jon Boise; Godfoy, Thomas

    2012-01-01

    This paper summarizes the progress that has been made in the development of the Fission Power System Technology Demonstration Unit (TDU). The reactor simulator core and Annular Linear Induction Pump have been fabricated and assembled into a test loop at the NASA Marshall Space Flight Center. A 12 kWe Power Conversion Unit (PCU) is being developed consisting of two 6 kWe free-piston Stirling engines. The two 6 kWe engines have been fabricated by Sunpower Inc. and are currently being tested separately prior to integration into the PCU. The Facility Cooling System (FCS) used to reject convertor waste heat has been assembled and tested at the NASA Glenn Research Center (GRC). The structural elements, including a Buildup Assembly Platform (BAP) and Upper Truss Structure (UTS) have been fabricated, and will be used to test cold-end components in thermal vacuum prior to TDU testing. Once all components have been fully tested at the subsystem level, they will be assembled into an end-to-end system and tested in thermal vacuum at GRC.

  14. Development Status of the Fission Power System Technology Demonstration Unit

    NASA Technical Reports Server (NTRS)

    Briggs, Maxwell H.; Gibson, Marc A.; Geng, Steven M; Pearson, Jon Boise; Godfroy, Thomas

    2012-01-01

    This paper summarizes the progress that has been made in the development of the Fission Power System Technology Demonstration Unit (TDU). The reactor simulator core and Annular Linear Induction Pump have been fabricated and assembled into a test loop at the NASA Marshall Space Flight Center. A 12 kWe Power Conversion Unit (PCU) is being developed consisting of two 6 kWe free-piston Stirling engines. The two 6 kWe engines have been fabricated by Sunpower Inc. and are currently being tested separately prior to integration into the PCU. The Facility Cooling System (FCS) used to reject convertor waste heat has been assembled and tested at the NASA Glenn Research Center (GRC). The structural elements, including a Buildup Assembly Platform (BAP) and Upper Truss Structure (UTS) have been fabricated, and will be used to test cold-end components in thermal vacuum prior to TDU testing. Once all components have been fully tested at the subsystem level, they will be assembled into an end-to-end system and tested in thermal vacuum at NASA GRC.

  15. Secure, Mobile, Wireless Network Technology Designed, Developed, and Demonstrated

    NASA Technical Reports Server (NTRS)

    Ivancic, William D.; Paulsen, Phillip E.

    2004-01-01

    The inability to seamlessly disseminate data securely over a high-integrity, wireless broadband network has been identified as a primary technical barrier to providing an order-of-magnitude increase in aviation capacity and safety. Secure, autonomous communications to and from aircraft will enable advanced, automated, data-intensive air traffic management concepts, increase National Air Space (NAS) capacity, and potentially reduce the overall cost of air travel operations. For the first time ever, secure, mobile, network technology was designed, developed, and demonstrated with state-ofthe- art protocols and applications by a diverse, cooperative Government-industry team led by the NASA Glenn Research Center. This revolutionary technology solution will make fundamentally new airplane system capabilities possible by enabling secure, seamless network connections from platforms in motion (e.g., cars, ships, aircraft, and satellites) to existing terrestrial systems without the need for manual reconfiguration. Called Mobile Router, the new technology autonomously connects and configures networks as they traverse from one operating theater to another. The Mobile Router demonstration aboard the Neah Bay, a U.S. Coast Guard vessel stationed in Cleveland, Ohio, accomplished secure, seamless interoperability of mobile network systems across multiple domains without manual system reconfiguration. The Neah Bay was chosen because of its low cost and communications mission similarity to low-Earth-orbiting satellite platforms. This technology was successfully advanced from technology readiness level (TRL) 2 (concept and/or application formation) to TRL 6 (system model or prototype demonstration in a relevant environment). The secure, seamless interoperability offered by the Mobile Router and encryption device will enable several new, vehicle-specific and systemwide technologies to perform such things as remote, autonomous aircraft performance monitoring and early detection and

  16. Reactive Leadership: Divining, Developing, and Demonstrating Community Ontologies

    NASA Astrophysics Data System (ADS)

    Graybeal, J.

    2008-12-01

    The Marine Metadata Interoperability Project (known as MMI, on the web at http://marinemetadata.org) was formed to provide leadership in metadata practices to the marine science community. In 2004 this meant finding and writing about resources and best practices, which until then were all but invisible. In 2008 the scope is far wider, encompassing comprehensive guidance, collaborative community environments, and introduction and demonstration of advanced technologies to an increasingly interested scientific domain. MMI's technical leadership, based on experiences gained in the hydrologic community, emphasized the role ontologies could play in marine science. An early MMI workshop successfully incorporated a large number of community vocabularies, tools to harmonize them in a common ontological format, and the mapping of terms from vocabularies expressed in that format. That 2005 workshop demonstrated the connections to be made among different community vocabularies, and was well regarded by participants, but did not lead to widespread adoption of the tools, technologies, or even the vocabularies. Ontology development efforts for marine sensors and platforms showed intermittent progress, but again were not adopted or pushed toward completion. It is now 2008, and the marine community is increasingly attentive to a wide range of interoperability issues. A large part of the community has at least heard of "semantic interoperability", and many understand its critical role in finding and working with data. Demand for specific solutions, and for workable approaches, is becoming more vocal in the marine community. Yet there is still no encompassing model in place for achieving semantic interoperability, only simple operational registries have been set up for oceanographic community vocabularies, and only a few isolated applications demonstrate how semantic barriers can be overcome. Why has progress been so slow? Are good answers on the horizon? And if we build it, will the

  17. Development and Demonstration of Active Noise Control Concepts

    NASA Technical Reports Server (NTRS)

    Kraft, R.; Hu, Z.; Sommerfeldt, S.; Walker, B.; Hersh, A.; Luo, H.; Spencer, M.; Hallman, D.; Mitchell, C.; Sutliff, D.

    2000-01-01

    This report details design methods for and feasibility of an Active Noise Control (ANC) system using flush-wall-mounted sensors and actuators to reduce turbofan engine rotor-stator interaction noise. ANC concepts capable of suppressing discrete-tone spinning modes containing several cut-on radial mode were identified, developed analytically, and evaluated. Separate ANC systems that suppressed at least three radial modes in a cylindrical inlet duct and three radial modes in an exhaust annulus were developed. These designs resulted in inlet duct and exhaust duct tests that were performed at NASA on the 4-ft ANC Fan in the NASA Glenn AAPL facility. Effective suppression of 2-BPF spinning mode m = 2 tone noise was achieved over a range of fan speeds 1800 to 2450 rpm, where up to 4 radials were present. In the inlet duct, up to 12 dB reduction was obtained for 3 radial modes, and up to 4 dB was obtained with 4 radial modes. In the exhaust duct, up to 15 dB PWL reduction was obtained with either two or three radial modes present. Thus, the ability to suppress multiple radial modes for tones in both the inlet and exhaust ducts has been successfully demonstrated. Implications of ANC system design requirements on installation and system integration issues for ANC systems capable of suppressing higher order radial mode content when applied to a 767 using twin CF6 engines were evaluated analytically. The analytical results indicated an ANC system must be part of an integrated design to be effective.

  18. WhiD and WhiB, Homologous Proteins Required for Different Stages of Sporulation in Streptomyces coelicolor A3(2)

    PubMed Central

    Molle, Virginie; Palframan, Wendy J.; Findlay, Kim C.; Buttner, Mark J.

    2000-01-01

    The whiD locus, which is required for the differentiation of Streptomyces coelicolor aerial hyphae into mature spore chains, was localized by map-based cloning to the overlap between cosmids 6G4 and D63 of the minimal ordered library of Redenbach et al. (M. Redenbach et al., Mol. Microbiol. 21:77–96, 1996). Subcloning and sequencing showed that whiD encodes a homologue of WhiB, a protein required for the initiation of sporulation septation in S. coelicolor. WhiD and WhiB belong to a growing family of small (76- to 112-residue) proteins of unknown biochemical function in which four cysteines are absolutely conserved; all known members of this family are found in the actinomycetes. A constructed whiD null mutant showed reduced levels of sporulation, and those spores that did form were heat sensitive, lysed extensively, and were highly irregular in size, arising at least in part from irregularity in septum placement. The whiD null mutant showed extreme variation in spore cell wall deposition; most spores had uniformly thin (20- to 30-nm) walls, but spore chains were frequently observed in which there was irregular but very pronounced (up to 170 nm) cell wall thickening at the junctions between spores. whiD null mutant spores were frequently partitioned into irregular smaller units through the deposition of additional septa, which were often laid down in several different planes, very close to the spore poles. These “minicompartments” appeared to be devoid of chromosomal DNA. Two whiD promoters, whiDp1 and whiDp2, were identified, and their activities were analyzed during development of wild-type S. coelicolor on solid medium. Both promoters were developmentally regulated; whiDp1 and whiDp2 transcripts were detected transiently, approximately at the time when sporulation septa were observed in the aerial hyphae. PMID:10671449

  19. Conformal window manufacturing process development and demonstration for polycrystalline materials

    NASA Astrophysics Data System (ADS)

    Smith, Nathan E.; Gould, Alan R.; Hordin, Tom; Medicus, Kate; Walters, Mark; Brophy, Matthew; DeGroote Nelson, Jessica

    2013-09-01

    Conformal windows pose new and unique challenges to manufacturing due to the shape, measurement of, and requested hard polycrystalline materials. Their non-rotationally symmetric shape and high departure surfaces do not lend themselves to traditional optical fabrication processes. The hard crystalline materials are another challenge due to increased processing time and possibility of grain decoration. We have developed and demonstrated a process for manufacturing various conformal windows out of fused silica, glass, zinc-sulfide multispectral, and spinel. The current process involves CNC generation/grinding, VIBE polishing, and sub-aperture figure correction. The CNC generation step incorporates an ultrasonic assisted grinding machine; the machine settings and tool are being continuously optimized for minimal sub-surface damage and surface form error. In VIBE, polishing to less than 5 nm rms surface roughness while maintaining overall form error is accomplished with a full aperture conformal polishing tool and with rapid removal rates. The final sub-aperture polishing step corrects the overall form error. Currently we utilize our CMM for surface form measurement and have shown that we can produce spinel conformal windows with form error within +/-10 micrometers of the nominal shape, without grain decoration. This conformal window manufacturing process is continuously optimized for cost reduction and precision of the final optic.

  20. Adenosine deaminase from Streptomyces coelicolor: recombinant expression, purification and characterization.

    PubMed

    Pornbanlualap, Somchai; Chalopagorn, Pornchanok

    2011-08-01

    The sequencing of the genome of Streptomyces coelicolor A3(2) identified seven putative adenine/adenosine deaminases and adenosine deaminase-like proteins, none of which have been biochemically characterized. This report describes recombinant expression, purification and characterization of SCO4901 which had been annotated in data bases as a putative adenosine deaminase. The purified putative adenosine deaminase gives a subunit Mr=48,400 on denaturing gel electrophoresis and an oligomer molecular weight of approximately 182,000 by comparative gel filtration. These values are consistent with the active enzyme being composed of four subunits with identical molecular weights. The turnover rate of adenosine is 11.5 s⁻¹ at 30 °C. Since adenine is deaminated ∼10³ slower by the enzyme when compared to that of adenosine, these data strongly show that the purified enzyme is an adenosine deaminase (ADA) and not an adenine deaminase (ADE). Other adenine nucleosides/nucleotides, including 9-β-D-arabinofuranosyl-adenine (ara-A), 5'-AMP, 5'-ADP and 5'-ATP, are not substrates for the enzyme. Coformycin and 2'-deoxycoformycin are potent competitive inhibitors of the enzyme with inhibition constants of 0.25 and 3.4 nM, respectively. Amino acid sequence alignment of ScADA with ADAs from other organisms reveals that eight of the nine highly conserved catalytic site residues in other ADAs are also conserved in ScADA. The only non-conserved residue is Asn317, which replaces Asp296 in the murine enzyme. Based on these data, it is suggested here that ADA and ADE proteins are divergently related enzymes that have evolved from a common α/β barrel scaffold to catalyze the deamination of different substrates, using a similar catalytic mechanism. PMID:21511036

  1. Early Development of Demonstratives in Pre-Qin Chinese

    ERIC Educational Resources Information Center

    Deng, Lin

    2011-01-01

    This dissertation offers a new dynamic account of the evolution of the demonstrative system in pre-Qin Chinese based on a comprehensive linguistic analysis of the phonological, morphological, syntactic, semantic, and pragmatic aspects of demonstratives attested in two corpora of excavated texts, i.e. the oracle-bone inscriptions dated to the late…

  2. RNA-Seq analysis reveals a six-gene SoxR regulon in Streptomyces coelicolor.

    PubMed

    Naseer, Nawar; Shapiro, Joshua A; Chander, Monica

    2014-01-01

    The redox-regulated transcription factor SoxR is conserved in diverse bacteria, but emerging studies suggest that this protein plays distinct physiological roles in different bacteria. SoxR regulates a global oxidative stress response (involving > 100 genes) against exogenous redox-cycling drugs in Escherichia coli and related enterics. In the antibiotic producers Streptomyces coelicolor and Pseudomonas aeruginosa, however, SoxR regulates a smaller number of genes that encode membrane transporters and proteins with homology to antibiotic-tailoring enzymes. In both S. coelicolor and P. aeruginosa, SoxR-regulated genes are expressed in stationary phase during the production of endogenously-produced redox-active antibiotics. These observations suggest that SoxR evolved to sense endogenous secondary metabolites and activate machinery to process and transport them in antibiotic-producing bacteria. Previous bioinformatics analysis that searched the genome for SoxR-binding sites in putative promoters defined a five-gene SoxR regulon in S. coelicolor including an ABC transporter, two oxidoreductases, a monooxygenase and an epimerase/dehydratase. Since this in silico screen may have missed potential SoxR-targets, we conducted a whole genome transcriptome comparison of wild type S. coelicolor and a soxR-deficient mutant in stationary phase using RNA-Seq. Our analysis revealed a sixth SoxR-regulated gene in S. coelicolor that encodes a putative quinone oxidoreductase. Knowledge of the full complement of genes regulated by SoxR will facilitate studies to elucidate the function of this regulatory molecule in antibiotic producers. PMID:25162599

  3. Development requirements for a successful ground based CELSS demonstration

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Considerations critical to a ground based control demonstration were identified. The controlled ecological life support system technologies were assessed for nutrition and food processing, food production, waste processing, and systems engineering/modeling.

  4. Genome-Wide Analysis of In Vivo Binding of the Master Regulator DasR in Streptomyces coelicolor Identifies Novel Non-Canonical Targets

    PubMed Central

    Świątek-Połatyńska, Magdalena A.; Bucca, Giselda; Laing, Emma; Gubbens, Jacob; Titgemeyer, Fritz; Smith, Colin P.; Rigali, Sébastien; van Wezel, Gilles P.

    2015-01-01

    Streptomycetes produce a wealth of natural products, including over half of all known antibiotics. It was previously demonstrated that N-acetylglucosamine and secondary metabolism are closely entwined in streptomycetes. Here we show that DNA recognition by the N-acetylglucosamine-responsive regulator DasR is growth-phase dependent, and that DasR can bind to sites in the S. coelicolor genome that have no obvious resemblance to previously identified DasR-responsive elements. Thus, the regulon of DasR extends well beyond what was previously predicted and includes a large number of genes with functions far removed from N-acetylglucosamine metabolism, such as genes for small RNAs and DNA transposases. Conversely, the DasR regulon during vegetative growth largely correlates to the presence of canonical DasR-responsive elements. The changes in DasR binding in vivo following N-acetylglucosamine induction were studied in detail and a possible molecular mechanism by which the influence of DasR is extended is discussed. Discussion of DasR binding was further informed by a parallel transcriptome analysis of the respective cultures. Evidence is provided that DasR binds directly to the promoters of all genes encoding pathway-specific regulators of antibiotic production in S. coelicolor, thereby providing an exquisitely simple link between nutritional control and secondary metabolism. PMID:25875084

  5. Structural and functional basis of transcriptional regulation by TetR family protein CprB from S. coelicolor A3(2)

    PubMed Central

    Bhukya, Hussain; Bhujbalrao, Ruchika; Bitra, Aruna; Anand, Ruchi

    2014-01-01

    Antibiotic production and resistance pathways in Streptomyces are dictated by the interplay of transcriptional regulatory proteins that trigger downstream responses via binding to small diffusible molecules. To decipher the mode of DNA binding and the associated allosteric mechanism in the sub-class of transcription factors that are induced by γ-butyrolactones, we present the crystal structure of CprB in complex with the consensus DNA element to a resolution of 3.25 Å. Binding of the DNA results in the restructuring of the dimeric interface of CprB, inducing a pendulum-like motion of the helix-turn-helix motif that inserts into the major groove. The crystal structure revealed that, CprB is bound to DNA as a dimer of dimers with the mode of binding being analogous to the broad spectrum multidrug transporter protein QacR from the antibiotic resistant strain Staphylococcus aureus. It was demonstrated that the CprB displays a cooperative mode of DNA binding, following a clamp and click model. Experiments performed on a subset of DNA sequences from Streptomyces coelicolor A3(2) suggest that CprB is most likely a pleiotropic regulator. Apart from serving as an autoregulator, it is potentially a part of a network of proteins that modulates the γ-butyrolactone synthesis and antibiotic regulation pathways in S. coelicolor A3(2). PMID:25092919

  6. RNA Polymerase Sigma Factor That Blocks Morphological Differentiation by Streptomyces coelicolor

    PubMed Central

    Gehring, Amy M.; Yoo, Narie J.; Losick, Richard

    2001-01-01

    The filamentous bacterium Streptomyces coelicolor undergoes a complicated process of morphological differentiation that begins with the formation of an aerial mycelium and culminates in sporulation. Genes required for the initiation of aerial mycelium formation have been termed bld (bald), describing the smooth, undifferentiated colonies of mutant strains. By using an insertional mutagenesis protocol that relies on in vitro transposition, we have isolated a bld mutant harboring an insertion in a previously uncharacterized gene, SCE59.12c, renamed here rsuA. The insertion mutant exhibited no measurable growth defect but failed to produce an aerial mycelium and showed a significant delay in the production of the polyketide antibiotic actinorhodin. The rsuA gene encodes an apparent anti-sigma factor and is located immediately downstream of SCE59.13c, renamed here sigU, whose product is inferred to be a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors. The absence of rsuA in a strain that contained sigU caused a block in development, and the overexpression of sigU in an otherwise wild-type strain caused a delay in aerial mycelium formation. However, a strain in which both rsuA and sigU had been deleted was able to undergo morphological differentiation normally. We conclude that the rsuA-encoded anti-sigma factor is responsible for antagonizing the function of the sigma factor encoded by sigU. We also conclude that the sigU-encoded sigma factor is not normally required for development but that its uncontrolled activity obstructs morphological differentiation at an early stage. PMID:11566999

  7. Adventure Central: Applying the "Demonstration Plot" Concept to Youth Development

    ERIC Educational Resources Information Center

    Cochran, Graham; Arnett, Nate; Ferrari, Theresa M.

    2006-01-01

    Historically, land-grant university research stations focused on demonstrating successful farming methods. The land-grant mission and the principles at its foundation have broad applicability, and we believe the land-grant principles can be successfully applied in urban settings to a university's work with youth and families. In this article we…

  8. Oxygen and Nitrate Respiration in Streptomyces coelicolor A3(2).

    PubMed

    Sawers, R G; Falke, D; Fischer, M

    2016-01-01

    Streptomyces species belong to the phylum Actinobacteria and can only grow with oxygen as a terminal electron acceptor. Like other members of this phylum, such as corynebacteria and mycobacteria, the aerobic respiratory chain lacks a soluble cytochrome c. It is therefore implicit that direct electron transfer between the cytochrome bc1 and the cytochrome aa3 oxidase complexes occurs. The complex developmental cycle of streptomycetes manifests itself in the production of spores, which germinate in the presence of oxygen into a substrate mycelium that greatly facilitates acquisition of nutrients necessary to support their saprophytic lifestyle in soils. Due to the highly variable oxygen levels in soils, streptomycetes have developed means of surviving long periods of hypoxia or even anaerobiosis but they fail to grow under these conditions. Little to nothing is understood about how they maintain viability under conditions of oxygen limitation. It is assumed that they can utilise a number of different electron acceptors to help them maintain a membrane potential, one of which is nitrate. The model streptomycete remains Streptomyces coelicolor A3(2), and it synthesises three nonredundant respiratory nitrate reductases (Nar). These Nar enzymes are synthesised during different phases of the developmental cycle and they are functional only under oxygen-limiting (<5% oxygen in air) conditions. Nevertheless, the regulation of their synthesis does not appear to be responsive to nitrate and in the case of Nar1, it appears to be developmentally regulated. This review highlights some of the novel aspects of our current, but somewhat limited, knowledge of respiration in these fascinating bacteria. PMID:27134020

  9. Redox flow cell development and demonstration project, calendar year 1977

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Research and development on the redox flow cell conducted from January 1, 1977, to December 31, 1977, are described in this report. The major focus of the effort during 1977 was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable ion exchange membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  10. DEMONSTRATION BULLETIN: FLAME REACTOR - HORSEHEAD RESOURCE DEVELOPMENT COMPANY, INC.

    EPA Science Inventory

    The Horsehead Resource Development Company, Inc. (HRD) Flame Reactor is a patented and proven high temperature thermal process designed to safely treat industrial residues and wastes containing metals. During processing, the waste material is introduced into the hottest portio...

  11. Redox flow cell development and demonstration project, calendar year 1976

    NASA Technical Reports Server (NTRS)

    1977-01-01

    The major focus of the effort was the key technology issues that directly influence the fundamental feasibility of the overall redox concept. These issues were the development of a suitable semipermeable separator membrane for the system, the screening and study of candidate redox couples to achieve optimum cell performance, and the carrying out of systems analysis and modeling to develop system performance goals and cost estimates.

  12. Flexible structure control laboratory development and technology demonstration

    NASA Technical Reports Server (NTRS)

    Vivian, H. C.; Blaire, P. E.; Eldred, D. B.; Fleischer, G. E.; Ih, C.-H. C.; Nerheim, N. M.; Scheid, R. E.; Wen, J. T.

    1987-01-01

    An experimental structure is described which was constructed to demonstrate and validate recent emerging technologies in the active control and identification of large flexible space structures. The configuration consists of a large, 20 foot diameter antenna-like flexible structure in the horizontal plane with a gimballed central hub, a flexible feed-boom assembly hanging from the hub, and 12 flexible ribs radiating outward. Fourteen electrodynamic force actuators mounted to the hub and to the individual ribs provide the means to excite the structure and exert control forces. Thirty permanently mounted sensors, including optical encoders and analog induction devices provide measurements of structural response at widely distributed points. An experimental remote optical sensor provides sixteen additional sensing channels. A computer samples the sensors, computes the control updates and sends commands to the actuators in real time, while simultaneously displaying selected outputs on a graphics terminal and saving them in memory. Several control experiments were conducted thus far and are documented. These include implementation of distributed parameter system control, model reference adaptive control, and static shape control. These experiments have demonstrated the successful implementation of state-of-the-art control approaches using actual hardware.

  13. Carbon-Flux Distribution within Streptomyces coelicolor Metabolism: A Comparison between the Actinorhodin-Producing Strain M145 and Its Non-Producing Derivative M1146

    PubMed Central

    Coze, Fabien; Gilard, Françoise; Tcherkez, Guillaume; Virolle, Marie-Joëlle; Guyonvarch, Armel

    2013-01-01

    Metabolic Flux Analysis is now viewed as essential to elucidate the metabolic pattern of cells and to design appropriate genetic engineering strategies to improve strain performance and production processes. Here, we investigated carbon flux distribution in two Streptomyces coelicolor A3 (2) strains: the wild type M145 and its derivative mutant M1146, in which gene clusters encoding the four main antibiotic biosynthetic pathways were deleted. Metabolic Flux Analysis and 13C-labeling allowed us to reconstruct a flux map under steady-state conditions for both strains. The mutant strain M1146 showed a higher growth rate, a higher flux through the pentose phosphate pathway and a higher flux through the anaplerotic phosphoenolpyruvate carboxylase. In that strain, glucose uptake and the flux through the Krebs cycle were lower than in M145. The enhanced flux through the pentose phosphate pathway in M1146 is thought to generate NADPH enough to face higher needs for biomass biosynthesis and other processes. In both strains, the production of NADPH was higher than NADPH needs, suggesting a key role for nicotinamide nucleotide transhydrogenase for redox homeostasis. ATP production is also likely to exceed metabolic ATP needs, indicating that ATP consumption for maintenance is substantial.Our results further suggest a possible competition between actinorhodin and triacylglycerol biosynthetic pathways for their common precursor, acetyl-CoA. These findings may be instrumental in developing new strategies exploiting S. coelicolor as a platform for the production of bio-based products of industrial interest. PMID:24376790

  14. Development and demonstration of flueric sounding rocket motor ignition

    NASA Technical Reports Server (NTRS)

    Marchese, V. P.

    1974-01-01

    An analytical and experimental program is described which established a flueric rocket motor ignition system concept incorporating a pneumatic match with a simple hand pump as the only energy source. An evaluation was made of this concept to determine the margins of the operating range and capabilities of every component of the system. This evaluation included a determination of power supply requirements, ignitor geometry and alinement, ignitor/propellant interfacing and materials and the effects of ambient temperatures and pressure. It was demonstrated that an operator using a simple hand pump for 30 seconds could ignite BKNO3 at a standoff distance of 100 m (330 ft) with the only connection to the ignitor being a piece of plastic pneumatic tubing.

  15. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... include such terms and conditions at least as protective as the criteria for municipal solid waste... and quantities of municipal solid waste and non-hazardous wastes which the State Director...

  16. 40 CFR 258.4 - Research, development, and demonstration permits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ...) SOLID WASTES CRITERIA FOR MUNICIPAL SOLID WASTE LANDFILLS General § 258.4 Research, development, and... include such terms and conditions at least as protective as the criteria for municipal solid waste... and quantities of municipal solid waste and non-hazardous wastes which the State Director...

  17. DESIGN, DEVELOPMENT, AND DEMONSTRATION OF A FINE PARTICULATE MEASURING DEVICE

    EPA Science Inventory

    The report describes the design, development, and testing of a fine particulate source monitoring instrument for real-time measurement of mass concentration as a function of aerodynamic particle size. It includes a literature review and selection of the operating principle on whi...

  18. Research, Development, Demonstration and Deployment Issues in the Power Sector

    EPA Science Inventory

    For Frank Princiotta’s book, Global Climate Change—The Technology Challenge In this chapter we explore the challenges in developing and deploying technology for mitigation of CO2 emissions associated with power generation. Past successes with controlling other pollutants (notab...

  19. POULTRY EXCRETA DEHYDRATION AND UTILIZATION: SYSTEM DEVELOPMENT AND DEMONSTRATION

    EPA Science Inventory

    A manure handling and drying system involving caged layers with daily manure collection, air drying, and dehydration in a flash-type dryer has been studied. Objectives of the study were to: (1) Develop a complete manure handling system to maximize pollution control; (2) determine...

  20. Prospects for international cooperation in SPS research, development and demonstration

    NASA Astrophysics Data System (ADS)

    Williamson, R. A.; Wasserman, A.

    1982-09-01

    Positive and negative aspects of international cooperation for the development of SPS satellites are discussed. The points raised are a result of a review by the U.S. Office of Technology Assessment, which projected a possible worldwide demand for 295-465 GW of SPS-derived electricity by the year 2030. The visibility of an SPS in the sky could possibly be a significant psychological factor for developing nations, although it may be impossible for any one nation to construct an SPS unilaterally. Sharing the expenses and resources among nations is calculated to make the construction of an SPS feasible. Member nations of the construction team would share in the power produced. Measures to avoid interference with other nations' spacecraft are noted to be necessary, as well as international agreements which prohibit antisatellite weapons. Attention is given to a distribution of research, the sharing of research data, and the sharing of financial burdens.

  1. Development and Flight Demonstration of Space Debris Monitor (SDM)

    NASA Astrophysics Data System (ADS)

    Kitazawa, Yukihito; Hanada, Toshiya; Matsumoto, Haruhisa; Kobayashi, Masanori; Sakurai, Akira; Yasaka, Tetsuo; Funakoshi, Kunihiro; Hasegawa, Sunao; Akahoshi, Yasuhiro; Kimoto, Yugo; Okudaira, Osamu; Kamiya, Koki; Nakamura, Maki

    2016-07-01

    The space debris monitor (SDM) is a large-area impact sensor for in situ measurements of micro-meteoroids and space debris of the sub-millimeter to millimeter size in the near-Earth space environment. These meteoroid and debris particles are very small to be detected by ground-based observations (radars and optical telescopes) but are sufficiently large to cause serious damage to spacecraft equipment in the low Earth orbit region. The nominal detection area of the SDM is 0.1 m^2 (0.35 m × 0.3 m), but its dimensions can be easily modified to accommodate different SDM constraints. The SDM is made from a flexible printed circuit, which is produced from a thin film of a nonconductive material (such as polyimide) on which thin conductive stripes are formed in parallel. The stripe width is approximately 50 μm, and the spatial separation is approximately 100 μm, as shown in Figure 1. When a micro-debris particle with an effective diameter near to or larger than the spatial separation of the stripes (here approximately 100 μm) collides with the sensor film at a velocity sufficient to penetrate it, one or more of the stripes are cut and become nonconductive. Debris impacts can thus be detected by monitoring the electrical conductivity (resistivity) of the stripes. This sensor system can measure the size of the incident micro-debris particles by detecting the number of severed stripes. The measurement concept is registered as a patent in many countries. The first SDM was launched with HTV-5 on August 19, 2015 and represented the world's first micro-debris measurement demonstration experiment to be conducted on the ISS using the concept of conductive (resistive) strip lines for real-time debris detection.

  2. Automotive Fuel Processor Development and Demonstration with Fuel Cell Systems

    SciTech Connect

    Nuvera Fuel Cells

    2005-04-15

    The potential for fuel cell systems to improve energy efficiency and reduce emissions over conventional power systems has generated significant interest in fuel cell technologies. While fuel cells are being investigated for use in many applications such as stationary power generation and small portable devices, transportation applications present some unique challenges for fuel cell technology. Due to their lower operating temperature and non-brittle materials, most transportation work is focusing on fuel cells using proton exchange membrane (PEM) technology. Since PEM fuel cells are fueled by hydrogen, major obstacles to their widespread use are the lack of an available hydrogen fueling infrastructure and hydrogen's relatively low energy storage density, which leads to a much lower driving range than conventional vehicles. One potential solution to the hydrogen infrastructure and storage density issues is to convert a conventional fuel such as gasoline into hydrogen onboard the vehicle using a fuel processor. Figure 2 shows that gasoline stores roughly 7 times more energy per volume than pressurized hydrogen gas at 700 bar and 4 times more than liquid hydrogen. If integrated properly, the fuel processor/fuel cell system would also be more efficient than traditional engines and would give a fuel economy benefit while hydrogen storage and distribution issues are being investigated. Widespread implementation of fuel processor/fuel cell systems requires improvements in several aspects of the technology, including size, startup time, transient response time, and cost. In addition, the ability to operate on a number of hydrocarbon fuels that are available through the existing infrastructure is a key enabler for commercializing these systems. In this program, Nuvera Fuel Cells collaborated with the Department of Energy (DOE) to develop efficient, low-emission, multi-fuel processors for transportation applications. Nuvera's focus was on (1) developing fuel processor

  3. Prioritizing orphan proteins for further study using phylogenomics and gene expression profiles in Streptomyces coelicolor

    PubMed Central

    2011-01-01

    Background Streptomyces coelicolor, a model organism of antibiotic producing bacteria, has one of the largest genomes of the bacterial kingdom, including 7825 predicted protein coding genes. A large number of these genes, nearly 34%, are functionally orphan (hypothetical proteins with unknown function). However, in gene expression time course data, many of these functionally orphan genes show interesting expression patterns. Results In this paper, we analyzed all functionally orphan genes of Streptomyces coelicolor and identified a list of "high priority" orphans by combining gene expression analysis and additional phylogenetic information (i.e. the level of evolutionary conservation of each protein). Conclusions The prioritized orphan genes are promising candidates to be examined experimentally in the lab for further characterization of their function. PMID:21899768

  4. [Determination of Streptomyces coelicolor A3(2) resistance to erythromycin].

    PubMed

    Puzynina, G G; Danilenko, V N; Vasil'chenko, L G; Mkrtumian, N M; Lomovskaia, N D

    1979-01-01

    Resistance to erythromycin is genetically unstable in strains of Streptomyces coelicolor A3(2). The frequent loss of resistance as well as reversion of sensitive variants to the original unstable resistance phenotype excluded the possibility that plasmid elimination is involved. The spontaneous frequency of occurrence of sensitive clones was 0.14 to 1.5%, the rate of reversion ranging from 1.10(-6) to 1.10(-8). Resistance to erythromycin has been mapped on the chromosomes of two S. coelicolor A3(2) derivatives in different sites: between markers adeC (v 10) and ArgA1 in the strain A617, between pheA1 and SCP1 in the strain S18. It is suggested that genetic instability of erythromycin resistance determinants having chromosomal location is due to transposition of genetic material. PMID:291565

  5. Tn4563 transposition in Streptomyces coelicolor and its application to isolation of new morphological mutants.

    PubMed Central

    Schauer, A T; Nelson, A D; Daniel, J B

    1991-01-01

    The Tn3-like transposon Tn4556 (and its derivatives Tn4560 and Tn4563) has been used for insertion mapping of genetic loci cloned on plasmids, but it has been difficult to obtain chromosomal insertions, largely because of the lack of a strong selection against transposon donor molecules. In this communication, we report two efficient selection techniques for transposition and their use in the isolation of chromosomal insertion mutations. A number of independent Streptomyces coelicolor morphological mutants (bld and whi) were obtained. Two of the bld mutations were mapped to locations on the chromosome by SCP1-mediated conjugation; at least one mutation, bld-5m1, appears to define a novel locus involved in control of S. coelicolor morphogenesis and antibiotic production. Images PMID:1650343

  6. Development and Demonstration of an Ada Test Generation System

    NASA Technical Reports Server (NTRS)

    1996-01-01

    important role in identifying testing requirements which are infeasible. This is especially true for data flow testing and modified condition/decision coverage. Our system uses in an essential way symbolic analysis and theorem proving technology, and we believe this work represents one of the few successful uses of a theorem prover working in a completely automatic fashion to solve a problem of practical interest. We believe this work anticipates an important trend away from purely syntactic-based methods for program analysis to semantic methods based on symbolic processing and inference technology. Other results demonstrating the practical use of automatic inference is being reported in hardware verification, although there are significant differences between the hardware work and ours. However, what is common and important is that general purpose theorem provers are being integrated with more special-purpose decision procedures to solve problems in analysis and verification. We are pursuina commercial opportunities for this work, and will use and extend the work in other projects we are engaged in. Ultimately we would like to rework the system to analyze C, C++, or Java as a key step toward commercialization.

  7. Purification of an Extracellular Signaling Molecule Involved in Production of Aerial Mycelium by Streptomyces coelicolor

    PubMed Central

    Nodwell, Justin R.; Losick, Richard

    1998-01-01

    We have extensively purified a factor from conditioned medium that restores aerial mycelium formation to a mutant of Streptomyces coelicolor that is defective in morphological differentiation. Response to this factor is shown to depend on the presence of the BldK oligopeptide import system. We suggest that this substance acts at the first step in a putative cascade of developmental regulatory signals. PMID:9495776

  8. Function and Evolution of Two Forms of SecDF Homologs in Streptomyces coelicolor

    PubMed Central

    Zhou, Zhan; Li, Yudong; Sun, Ning; Sun, Zhihao; Lv, Longxian; Wang, Yufeng; Shen, Libing; Li, Yong-Quan

    2014-01-01

    The general secretion (Sec) pathway plays a prominent role in bacterial protein export, and the accessory component SecDF has been shown to improve transportation efficiency. Inspection of Streptomyces coelicolor genome reveals the unexpected presence of two different forms of secDF homologous genes: one in fused form (secDF) and the other in separated form (secD and secF). However, the functional role of two SecDF homologs in S. coelicolor has not yet been determined. Transcriptional analysis of secDF homologs reveals that these genes are constitutively expressed. However, the transcript levels of secD and secF are much higher than that of secDF in S. coelicolor. Deletion of secDF or/and secD/secF in S. coelicolor did result in reduced secretion efficiency of Xylanase A and Amylase C, suggesting that they may have redundant functions for Sec-dependent translocation pathway. Moreover, our results also indicate that SecD/SecF plays a more prominent role than SecDF in protein translocation. Evolutionary analysis suggests that the fused and separated SecDF homologs in Streptomyces may have disparate evolutionary ancestries. SecD/SecF may be originated from vertical transmission of existing components from ancestor of Streptomyces species. However, SecDF may be derived from bacterial ancestors through horizontal gene transfer. Alternately, it is also plausible that SecDF may have arisen through additional gene duplication and fusion events. The acquisition of a second copy may confer a selective benefit to Streptomyces by enhancing protein transport capacity. Taken together, our results provide new insights into the potential biological function and evolutionary aspects of the prokaryotic SecDF complex. PMID:25140821

  9. Post-translational Serine/Threonine Phosphorylation and Lysine Acetylation: A Novel Regulatory Aspect of the Global Nitrogen Response Regulator GlnR in S. coelicolor M145

    PubMed Central

    Amin, Rafat; Franz-Wachtel, Mirita; Tiffert, Yvonne; Heberer, Martin; Meky, Mohamed; Ahmed, Yousra; Matthews, Arne; Krysenko, Sergii; Jakobi, Marco; Hinder, Markus; Moore, Jane; Okoniewski, Nicole; Maček, Boris; Wohlleben, Wolfgang; Bera, Agnieszka

    2016-01-01

    Soil-dwelling Streptomyces bacteria such as S.coelicolor have to constantly adapt to the nitrogen (N) availability in their habitat. Thus, strict transcriptional and post-translational control of the N-assimilation is fundamental for survival of this species. GlnR is a global response regulator that controls transcription of the genes related to the N-assimilation in S. coelicolor and other members of the Actinomycetales. GlnR represents an atypical orphan response regulator that is not activated by the phosphorylation of the conserved aspartate residue (Asp 50). We have applied transcriptional analysis, LC-MS/MS analysis and electrophoretic mobility shift assays (EMSAs) to understand the regulation of GlnR in S. coelicolor M145. The expression of glnR and GlnR-target genes was revisited under four different N-defined conditions and a complex N-rich condition. Although, the expression of selected GlnR-target genes was strongly responsive to changing N-concentrations, the glnR expression itself was independent of the N-availability. Using LC-MS/MSanalysis we demonstrated that GlnR was post-translationally modified. The post-translational modifications of GlnR comprise phosphorylation of the serine/threonine residues and acetylation of lysine residues. In the complex N-rich medium GlnR was phosphorylated on six serine/threonine residues and acetylated on one lysine residue. Under defined N-excess conditions only two phosphorylated residues were detected whereas under defined N-limiting conditions no phosphorylation was observed. GlnR phosphorylation is thus clearly correlated with N-rich conditions. Furthermore, GlnR was acetylated on four lysine residues independently of the N-concentration in the defined media and on only one lysine residue in the complex N-rich medium. Using EMSAs we demonstrated that phosphorylation inhibited the binding of GlnR to its targets genes, whereas acetylation had little influence on the formation of GlnR-DNA complex. This study clearly

  10. Post-translational Serine/Threonine Phosphorylation and Lysine Acetylation: A Novel Regulatory Aspect of the Global Nitrogen Response Regulator GlnR in S. coelicolor M145.

    PubMed

    Amin, Rafat; Franz-Wachtel, Mirita; Tiffert, Yvonne; Heberer, Martin; Meky, Mohamed; Ahmed, Yousra; Matthews, Arne; Krysenko, Sergii; Jakobi, Marco; Hinder, Markus; Moore, Jane; Okoniewski, Nicole; Maček, Boris; Wohlleben, Wolfgang; Bera, Agnieszka

    2016-01-01

    Soil-dwelling Streptomyces bacteria such as S.coelicolor have to constantly adapt to the nitrogen (N) availability in their habitat. Thus, strict transcriptional and post-translational control of the N-assimilation is fundamental for survival of this species. GlnR is a global response regulator that controls transcription of the genes related to the N-assimilation in S. coelicolor and other members of the Actinomycetales. GlnR represents an atypical orphan response regulator that is not activated by the phosphorylation of the conserved aspartate residue (Asp 50). We have applied transcriptional analysis, LC-MS/MS analysis and electrophoretic mobility shift assays (EMSAs) to understand the regulation of GlnR in S. coelicolor M145. The expression of glnR and GlnR-target genes was revisited under four different N-defined conditions and a complex N-rich condition. Although, the expression of selected GlnR-target genes was strongly responsive to changing N-concentrations, the glnR expression itself was independent of the N-availability. Using LC-MS/MSanalysis we demonstrated that GlnR was post-translationally modified. The post-translational modifications of GlnR comprise phosphorylation of the serine/threonine residues and acetylation of lysine residues. In the complex N-rich medium GlnR was phosphorylated on six serine/threonine residues and acetylated on one lysine residue. Under defined N-excess conditions only two phosphorylated residues were detected whereas under defined N-limiting conditions no phosphorylation was observed. GlnR phosphorylation is thus clearly correlated with N-rich conditions. Furthermore, GlnR was acetylated on four lysine residues independently of the N-concentration in the defined media and on only one lysine residue in the complex N-rich medium. Using EMSAs we demonstrated that phosphorylation inhibited the binding of GlnR to its targets genes, whereas acetylation had little influence on the formation of GlnR-DNA complex. This study clearly

  11. The development and testing of pulsed detonation engine ground demonstrators

    NASA Astrophysics Data System (ADS)

    Panicker, Philip Koshy

    2008-10-01

    The successful implementation of a PDE running on fuel and air mixtures will require fast-acting fuel-air injection and mixing techniques, detonation initiation techniques such as DDT enhancing devices or a pre-detonator, an effective ignition system that can sustain repeated firing at high rates and a fast and capable, closed-loop control system. The control system requires high-speed transducers for real-time monitoring of the PDE and the detection of the detonation wave speed. It is widely accepted that the detonation properties predicted by C-J detonation relations are fairly accurate in comparison to experimental values. The post-detonation flow properties can also be expressed as a function of wave speed or Mach number. Therefore, the PDE control system can use C-J relations to predict the post-detonation flow properties based on measured initial conditions and compare the values with those obtained from using the wave speed. The controller can then vary the initial conditions within the combustor for the subsequent cycle, by modulating the frequency and duty cycle of the valves, to obtain optimum air and fuel flow rates, as well as modulate the energy and timing of the ignition to achieve the required detonation properties. Five different PDE ground demonstrators were designed, built and tested to study a number of the required sub-systems. This work presents a review of all the systems that were tested, along with suggestions for their improvement. The PDE setups, ranged from a compact PDE with a 19 mm (3/4 in.) i.d., to two 25 mm (1 in.) i.d. setups, to a 101 mm (4 in.) i.d. dual-stage PDE setup with a pre-detonator. Propane-oxygen mixtures were used in the smaller PDEs. In the dual-stage PDE, propane-oxygen was used in the pre-detonator, while propane-air mixtures were used in the main combustor. Both rotary valves and solenoid valve injectors were studied. The rotary valves setups were tested at 10 Hz, while the solenoid valves were tested at up to 30 Hz

  12. Fertility properties and regulation of antimicrobial substance production by plasmid SCP2 of Streptomyces coelicolor.

    PubMed Central

    Troost, T R; Danilenko, V N; Lomovskaya, N D

    1979-01-01

    Streptomyces coelicolor A3(2) possesses two plasmids (SCP1 and SCP2) that act as sex factors. The plasmid deoxyribonucleic acid isolated from S. coelicolor A3(2) SCP1- strains A617 and A585 had the same molecular weight and endonuclease cleavage pattern as the SCP2 plasmid. The plasmidless strain S18 SCP2- was isolated from the A617 X A585 cross. SCP2 plasmid-containing strains acted as donors of chromosomal markers, whereas the plasmidless strain acted as recipient. The transfer of SCP2+ donor strain markers into the SCP2- recipient occurred at high frequencies (approximately 75%), was unidirectional, was initiated from a fixed region of the chromosome, and had the SCP2 fertility factor transferred first. The introduction of the SCP2 plasmid into a recipient strain greatly reduced the recombination frequency. These fertility properties differed from those previously reported, thereby suggesting that the SCP2 plasmid examined in this investigation may be an additional variant to those described in the literature. The SCP2 plasmid also regulated production of three antibacterial substances and conveyed resistance for S. coelicolor A3(2) strains against growth inhibition by one of them. Images PMID:500559

  13. Identification and physiological characterization of phosphatidic acid phosphatase enzymes involved in triacylglycerol biosynthesis in Streptomyces coelicolor

    PubMed Central

    2013-01-01

    Background Phosphatidic acid phosphatase (PAP, EC 3.1.3.4) catalyzes the dephosphorylation of phosphatidate yielding diacylglycerol (DAG), the lipid precursor for triacylglycerol (TAG) biosynthesis. Despite the importance of PAP activity in TAG producing bacteria, studies to establish its role in lipid metabolism have been so far restricted only to eukaryotes. Considering the increasing interest of bacterial TAG as a potential source of raw material for biofuel production, we have focused our studies on the identification and physiological characterization of the putative PAP present in the TAG producing bacterium Streptomyces coelicolor. Results We have identified two S. coelicolor genes, named lppα (SCO1102) and lppβ (SCO1753), encoding for functional PAP proteins. Both enzymes mediate, at least in part, the formation of DAG for neutral lipid biosynthesis. Heterologous expression of lppα and lppβ genes in E. coli resulted in enhanced PAP activity in the membrane fractions of the recombinant strains and concomitantly in higher levels of DAG. In addition, the expression of these genes in yeast complemented the temperature-sensitive growth phenotype of the PAP deficient strain GHY58 (dpp1lpp1pah1). In S. coelicolor, disruption of either lppα or lppβ had no effect on TAG accumulation; however, the simultaneous mutation of both genes provoked a drastic reduction in de novo TAG biosynthesis as well as in total TAG content. Consistently, overexpression of Lppα and Lppβ in the wild type strain of S. coelicolor led to a significant increase in TAG production. Conclusions The present study describes the identification of PAP enzymes in bacteria and provides further insights on the genetic basis for prokaryotic oiliness. Furthermore, this finding completes the whole set of enzymes required for de novo TAG biosynthesis pathway in S. coelicolor. Remarkably, the overexpression of these PAPs in Streptomyces bacteria contributes to a higher productivity of this single

  14. A novel Streptomyces gene, samR, with different effects on differentiation of Streptomyces ansochromogenes and Streptomyces coelicolor.

    PubMed

    Tan, Huarong; Tian, Yuqing; Yang, Haihua; Liu, Gang; Nie, Liping

    2002-03-01

    A 1.4-kb DNA fragment from Streptomyces ansochromogenes accelerated mycelium formation of S. ansochromogenes when present on a multicopy plasmid. The DNA fragment contains one complete open reading frame, designated samR, encoding a protein with 213 amino acids that contains a likely DNA-binding helix-turn-helix motif close to its N-terminus. The deduced SamR protein resembles the product of the hppR gene, which is involved in the regulation of catabolism of 3-(3-hydroxyphenyl) propionate in Rhodococcus globerulus. A samR disruption mutant was constructed that presented a bald phenotype and failed to form aerial hyphae and spores. We suggest that samR plays an important role in the emergence of aerial hyphae from substrate mycelium. An almost identical gene of Streptomyces coelicolor was also subjected to gene disruption. Surprisingly, the mutant was able to develop an aerial mycelium, but it remained white and deficient in sporulation instead of forming gray spores. PMID:11907684

  15. A sporulation-specific, sigF-dependent protein, SspA, affects septum positioning in Streptomyces coelicolor

    PubMed Central

    Tzanis, Angelos; Dalton, Kate A; Hesketh, Andrew; den Hengst, Chris D; Buttner, Mark J; Thibessard, Annabelle; Kelemen, Gabriella H

    2014-01-01

    The RNA polymerase sigma factor SigF controls late development during sporulation in the filamentous bacterium Streptomyces coelicolor. The only known SigF-dependent gene identified so far, SCO5321, is found in the biosynthetic cluster encoding spore pigment synthesis. Here we identify the first direct target for SigF, the gene sspA, encoding a sporulation-specific protein. Bioinformatic analysis suggests that SspA is a secreted lipoprotein with two PepSY signature domains. The sspA deletion mutant exhibits irregular sporulation septation and altered spore shape, suggesting that SspA plays a role in septum formation and spore maturation. The fluorescent translational fusion protein SspA–mCherry localized first to septum sites, then subsequently around the surface of the spores. Both SspA protein and sspA transcription are absent from the sigF null mutant. Moreover, in vitro transcription assay confirmed that RNA polymerase holoenzyme containing SigF is sufficient for initiation of transcription from a single sspA promoter. In addition, in vivo and in vitro experiments showed that sspA is a direct target of BldD, which functions to repress sporulation genes, including whiG, ftsZ and ssgB, during vegetative growth, co-ordinating their expression during sporulation septation. PMID:24261854

  16. In Vivo Analysis of HPr Reveals a Fructose-Specific Phosphotransferase System That Confers High-Affinity Uptake in Streptomyces coelicolor

    PubMed Central

    Nothaft, Harald; Parche, Stephan; Kamionka, Annette; Titgemeyer, Fritz

    2003-01-01

    HPr, the histidine-containing phosphocarrier protein of the bacterial phosphotransferase system (PTS), serves multiple functions in carbohydrate uptake and carbon source regulation in low-G+C-content gram-positive bacteria and in gram-negative bacteria. To assess the role of HPr in the high-G+C-content gram-positive organism Streptomyces coelicolor, the encoding gene, ptsH, was deleted. The ptsH mutant BAP1 was impaired in fructose utilization, while growth on other carbon sources was not affected. Uptake assays revealed that BAP1 could not transport appreciable amounts of fructose, while the wild type showed inducible high-affinity fructose transport with an apparent Km of 2 μM. Complementation and reconstitution experiments demonstrated that HPr is indispensable for a fructose-specific PTS activity. Investigation of the putative fruKA gene locus led to identification of the fructose-specific enzyme II permease encoded by the fruA gene. Synthesis of HPr was not specifically enhanced in fructose-grown cells and occurred also in the presence of non-PTS carbon sources. Transcriptional analysis of ptsH revealed two promoters that are carbon source regulated. In contrast to what happens in other bacteria, glucose repression of glycerol kinase was still operative in a ptsH background, which suggests that HPr is not involved in general carbon regulation. However, fructose repression of glycerol kinase was lost in BAP1, indicating that the fructose-PTS is required for transduction of the signal. This study provides the first molecular genetic evidence of a physiological role of the PTS in S. coelicolor. PMID:12533468

  17. Development and demonstration of manufacturing processes for fabricating graphite/LARC 160 polyimide structural elements

    NASA Technical Reports Server (NTRS)

    Frost, R. K.; Jones, J. S.; Dynes, P. J.; Wykes, D. H.

    1981-01-01

    The development and demonstration of manufacturing technologies for the structural application of Celion graphite/LARC-160 polyimide composite material is discussed. Process development and fabrication of demonstration components are discussed. Process development included establishing quality assurance of the basic composite material and processing, nondestructive inspection of fabricated components, developing processes for specific structural forms, and qualification of processes through mechanical testing. Demonstration components were fabricated. The demonstration components consisted of flat laminates, skin/stringer panels, honeycomb panels, chopped fiber compression moldings, and a technology demonstrator segment (TDS) representative of the space shuttle aft body flap.

  18. CDA is a new chromosomally-determined antibiotic from Streptomyces coelicolor A3(2).

    PubMed

    Hopwood, D A; Wright, H M

    1983-12-01

    Mutations (cda) leading to non-production of the new calcium-dependent antibiotic (CDA) of Streptomyces coelicolor A3(2) were closely linked on the chromosome. One representative mutation (cda-1) was mapped precisely between nicA and adeC. No cosynthesis of CDA was found in any pairwise combinations of 14 cda mutants. Mutations lacking aerial mycelium (bald mutations), mapping to the four previously described loci (bldA-D), were pleiotropically defective in production of CDA. PMID:6668466

  19. Resuscitation-Promoting Factors Are Cell Wall-Lytic Enzymes with Important Roles in the Germination and Growth of Streptomyces coelicolor

    PubMed Central

    Sexton, Danielle L.; St-Onge, Renée J.; Haiser, Henry J.; Yousef, Mary R.; Brady, Lauren; Gao, Chan; Leonard, Jacqueline

    2014-01-01

    Dormancy is a common strategy adopted by bacterial cells as a means of surviving adverse environmental conditions. For Streptomyces bacteria, this involves developing chains of dormant exospores that extend away from the colony surface. Both spore formation and subsequent spore germination are tightly controlled processes, and while significant progress has been made in understanding the underlying regulatory and enzymatic bases for these, there are still significant gaps in our understanding. One class of proteins with a potential role in spore-associated processes are the so-called resuscitation-promoting factors, or Rpfs, which in other actinobacteria are needed to restore active growth to dormant cell populations. The model species Streptomyces coelicolor encodes five Rpf proteins (RpfA to RfpE), and here we show that these proteins have overlapping functions during growth. Collectively, the S. coelicolor Rpfs promote spore germination and are critical for growth under nutrient-limiting conditions. Previous studies have revealed structural similarities between the Rpf domain and lysozyme, and our in vitro biochemical assays revealed various levels of peptidoglycan cleavage capabilities for each of these five Streptomyces enzymes. Peptidoglycan remodeling by enzymes such as these must be stringently governed so as to retain the structural integrity of the cell wall. Our results suggest that one of the Rpfs, RpfB, is subject to a unique mode of enzymatic autoregulation, mediated by a domain of previously unknown function (DUF348) located within the N terminus of the protein; removal of this domain led to significantly enhanced peptidoglycan cleavage. PMID:25512314

  20. Ergothioneine protects Streptomyces coelicolor A3(2) from oxidative stresses.

    PubMed

    Nakajima, Shunsuke; Satoh, Yasuharu; Yanashima, Kentaro; Matsui, Tomomi; Dairi, Tohru

    2015-09-01

    Thiol compounds with low-molecular weight, such as glutathione, mycothiol (MSH), bacillithiol, and ergothioneine (ERG), are known to protect microorganisms from oxidative stresses. Mycobacteria and actinobacteria utilize both MSH and ERG. The biological functions of MSH in mycobacteria have been extensively studied by genetic and biochemical studies, which have suggested it has critical roles for detoxification in cells. In contrast, the biological functions of ERG remain ambiguous because its biosynthetic genes were only recently identified in Mycobacterium avium. In this study, we constructed mutants of Streptomyces coelicolor A3(2), in which either the MSH or ERG biosynthetic gene was disrupted, and examined their phenotypes. A mshC (SCO1663)-disruptant completely lost MSH productivity. In contrast, a disruptant of the egtA gene (SCO0910) encoding γ-glutamyl-cysteine synthetase unexpectedly retained reduced productivity of ERG, probably because of the use of l-cysteine instead of γ-glutamyl-cysteine. Both disruptants showed delayed growth at the late logarithmic phase and were more susceptible to hydrogen peroxide and cumene hydroperoxide than the parental strain. Interestingly, the ERG-disruptant, which still kept reduced ERG productivity, was more susceptible. Furthermore, the ERG-disruptant accumulated 5-fold more MSH than the parental strain. In contrast, the amount of ERG was almost the same between the MSH-disruptant and the parental strain. Taken together, our results suggest that ERG is more important than MSH in S. coelicolor A3(2). PMID:25683449

  1. 18 CFR 367.1880 - Account 188, Research, development, or demonstration expenditures.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 18 Conservation of Power and Water Resources 1 2010-04-01 2010-04-01 false Account 188, Research... 188, Research, development, or demonstration expenditures. (a) This account must be charged with the cost of all expenditures coming within the meaning of research, development and demonstration (RD&D)...

  2. The two kinases, AbrC1 and AbrC2, of the atypical two-component system AbrC are needed to regulate antibiotic production and differentiation in Streptomyces coelicolor

    PubMed Central

    Rodríguez, Héctor; Rico, Sergio; Yepes, Ana; Franco-Echevarría, Elsa; Antoraz, Sergio; Santamaría, Ramón I.; Díaz, Margarita

    2015-01-01

    Two-component systems (TCSs) are the most important sensing mechanisms in bacteria. In Streptomyces, TCSs-mediated responses to environmental stimuli are involved in the regulation of antibiotic production. This study examines the individual role of two histidine kinases (HKs), AbrC1 and AbrC2, which form part of an atypical TCS in Streptomyces coelicolor. qRT-PCR analysis of the expression of both kinases demonstrated that both are expressed at similar levels in NB and NMMP media. Single deletion of abrC1 elicited a significant increase in antibiotic production, while deletion of abrC2 did not have any clear effect. The origin of this phenotype, probably related to the differential phosphorylation ability of the two kinases, was also explored indirectly, analyzing the toxic phenotypes associated with high levels of phosphorylated RR. The higher the AbrC3 regulator phosphorylation rate, the greater the cell toxicity. For the first time, the present work shows in Streptomyces the combined involvement of two different HKs in the response of a regulator to environmental signals. Regarding the possible applications of this research, the fact that an abrC1 deletion mutant overproduces three of the S. coelicolor antibiotics makes this strain an excellent candidate as a host for the heterologous production of secondary metabolites. PMID:26029189

  3. The two kinases, AbrC1 and AbrC2, of the atypical two-component system AbrC are needed to regulate antibiotic production and differentiation in Streptomyces coelicolor.

    PubMed

    Rodríguez, Héctor; Rico, Sergio; Yepes, Ana; Franco-Echevarría, Elsa; Antoraz, Sergio; Santamaría, Ramón I; Díaz, Margarita

    2015-01-01

    Two-component systems (TCSs) are the most important sensing mechanisms in bacteria. In Streptomyces, TCSs-mediated responses to environmental stimuli are involved in the regulation of antibiotic production. This study examines the individual role of two histidine kinases (HKs), AbrC1 and AbrC2, which form part of an atypical TCS in Streptomyces coelicolor. qRT-PCR analysis of the expression of both kinases demonstrated that both are expressed at similar levels in NB and NMMP media. Single deletion of abrC1 elicited a significant increase in antibiotic production, while deletion of abrC2 did not have any clear effect. The origin of this phenotype, probably related to the differential phosphorylation ability of the two kinases, was also explored indirectly, analyzing the toxic phenotypes associated with high levels of phosphorylated RR. The higher the AbrC3 regulator phosphorylation rate, the greater the cell toxicity. For the first time, the present work shows in Streptomyces the combined involvement of two different HKs in the response of a regulator to environmental signals. Regarding the possible applications of this research, the fact that an abrC1 deletion mutant overproduces three of the S. coelicolor antibiotics makes this strain an excellent candidate as a host for the heterologous production of secondary metabolites. PMID:26029189

  4. A comparison of key aspects of gene regulation in Streptomyces coelicolor and Escherichia coli using nucleotide-resolution transcription maps produced in parallel by global and differential RNA sequencing

    PubMed Central

    Romero, David A; Hasan, Ayad H; Lin, Yu-fei; Kime, Louise; Ruiz-Larrabeiti, Olatz; Urem, Mia; Bucca, Giselda; Mamanova, Lira; Laing, Emma E; van Wezel, Gilles P; Smith, Colin P; Kaberdin, Vladimir R; McDowall, Kenneth J

    2014-01-01

    Streptomyces coelicolor is a model for studying bacteria renowned as the foremost source of natural products used clinically. Post-genomic studies have revealed complex patterns of gene expression and links to growth, morphological development and individual genes. However, the underlying regulation remains largely obscure, but undoubtedly involves steps after transcription initiation. Here we identify sites involved in RNA processing and degradation as well as transcription within a nucleotide-resolution map of the transcriptional landscape. This was achieved by combining RNA-sequencing approaches suited to the analysis of GC-rich organisms. Escherichia coli was analysed in parallel to validate the methodology and allow comparison. Previously, sites of RNA processing and degradation had not been mapped on a transcriptome-wide scale for E. coli. Through examples, we show the value of our approach and data sets. This includes the identification of new layers of transcriptional complexity associated with several key regulators of secondary metabolism and morphological development in S. coelicolor and the identification of host-encoded leaderless mRNA and rRNA processing associated with the generation of specialized ribosomes in E. coli. New regulatory small RNAs were identified for both organisms. Overall the results illustrate the diversity in mechanisms used by different bacterial groups to facilitate and regulate gene expression. PMID:25266672

  5. Precision Information Environment (PIE) for International Safeguards: Pre-Demonstration Development Use Cases

    SciTech Connect

    Gastelum, Zoe N.; Henry, Michael J.

    2013-11-13

    In FY2013, the PIE International Safeguards team demonstrated our development progress to U.S. Department of Energy (DOE) staff from the Office of Nonproliferation and International Security (NA-24, our client) and the Office of Defense Nuclear Nonproliferation Research and Development (NA-22). Following the demonstration, the team was asked by our client to complete additional development prior to a planned demonstration at the International Atomic Energy Agency (IAEA), scheduled tentatively for January or spring of 2014. The team discussed four potential areas for development (in priority order), and will develop them as time and funding permit prior to an IAEA demonstration. The four capability areas are: 1. Addition of equipment manuals to PIE-accessible files 2. Optical character recognition (OCR) of photographed text 3. Barcode reader with information look-up from a database 4. Add Facilities to Data Model 5. Geospatial capabilities with information integration Each area will be described below in a use case.

  6. A Plan for the Development and Demonstration of Optical Communications for Deep Space

    NASA Technical Reports Server (NTRS)

    Lesh, J. R.; Deutsch, L. J.; Weber, W. J.

    1990-01-01

    In this article, an overall plan for the development and demonstration of optical communications for deep-space applications is presented. The current state of the technology for optical communications is presented. Then, the development and demonstration plan is presented in two parts: the overall major systems activities, followed by the generic technology developments that will enable them. The plan covers the path from laboratory subsystems demonstrations out to a full-scale flight experiment system for the proposed Mars Communications Relay Orbiter mission.

  7. A plan for the development and demonstration of optical communications for deep space

    NASA Technical Reports Server (NTRS)

    Lesh, James R.; Deutsch, Leslie J.; Weber, William J.

    1991-01-01

    A development history, current status, and prospective development account are given for deep-space optical communications, giving attention to overall systems-related activities and to the generic technology developments required for their implementation. An account is given of the progression envisioned from laboratory subsystems demonstrations to the full-scale flight experiment for the Mars Communications Relay Orbiter missions.

  8. TECHNOLOGY EVALUATION REPORT: SITE PROGRAM DEMONSTRATION TEST -HORSEHEAD RESEARCH DEVELOPMENT CO., INC. - FLAME REACTOR TECHNOLOGY

    EPA Science Inventory

    A SITE demonstration of the Horsehead Resource Development (HRD) Company, Inc. Flame Reactor Technology was conducted in March 1991 at the HRD facility in Monaca, Pennsylvania. or this demonstration, secondary lead smelter soda slag was treated to produce a potentially recyclable...

  9. Seeing Is Believing: Creating a Catalyst for Teacher Change through a Demonstration Classroom Professional Development Initiative

    ERIC Educational Resources Information Center

    Grierson, Arlene L.; Gallagher, Tiffany L.

    2009-01-01

    This qualitative case study explored the experiences of eight observing teachers, a demonstration teacher, and a program department consultant, who over a nine-month period participated in a demonstration classroom professional development initiative. Analyses of participants' experiences documented how the initiative created a catalyst for…

  10. ERDA/NASA-MSFC solar heating and cooling development and demonstration program

    NASA Technical Reports Server (NTRS)

    Price, J. M.

    1977-01-01

    The role of the Manned Space Flight Center (MSFC) in the National Solar Heating and Cooling program is evaluated. The responsibilities of the MSFC within this program are (1) to manage the development and testing of solar heating and cooling systems leading to marketable products, (2) to develop data acquisition equipment supporting a national data program, and (3) to manage and collect data from commercial demonstration sites. MSFC test facilities and demonstration sites are described.

  11. Interspecies modulation of bacterial development through iron competition and siderophore piracy

    PubMed Central

    Traxler, Matthew F.; Seyedsayamdost, Mohammad R.; Clardy, Jon; Kolter, Roberto

    2012-01-01

    Summary While soil-dwelling actinomycetes are renowned for secreting natural products, little is known about the roles of these molecules in mediating actinomycete interactions. In a previous co-culture screen, we found that one actinomycete, Amycolatopsis sp. AA4, inhibited aerial hyphae formation in adjacent colonies of Streptomyces coelicolor. A siderophore, amychelin, mediated this developmental arrest. Here we present genetic evidence that confirms the role of the amc locus in the production of amychelin and in the inhibition of S. coelicolor development. We further characterize the Amycolatopsis sp. AA4 - S. coelicolor interaction by examining expression of developmental and iron acquisition genes over time in co-culture. Manipulation of iron availability and/or growth near Amycolatopsis sp. AA4 led to alterations in expression of the critical developmental gene bldN, and other key down-stream genes in the S. coelicolor transcriptional cascade. In Amycolatopsis sp. AA4, siderophore genes were down-regulated when grown near S. coelicolor, leading us to find that deferrioxamine E, produced by S. coelicolor, could be readily utilized by Amycolatopsis sp. AA4. Collectively these results suggest that competition for iron via siderophore piracy and species-specific siderophores can alter patterns of gene expression and morphological differentiation during actinomycete interactions. PMID:22931126

  12. Interspecies modulation of bacterial development through iron competition and siderophore piracy.

    PubMed

    Traxler, Matthew F; Seyedsayamdost, Mohammad R; Clardy, Jon; Kolter, Roberto

    2012-11-01

    While soil-dwelling actinomycetes are renowned for secreting natural products, little is known about the roles of these molecules in mediating actinomycete interactions. In a previous co-culture screen, we found that one actinomycete, Amycolatopsis sp. AA4, inhibited aerial hyphae formation in adjacent colonies of Streptomyces coelicolor. A siderophore, amychelin, mediated this developmental arrest. Here we present genetic evidence that confirms the role of the amc locus in the production of amychelin and in the inhibition of S. coelicolor development. We further characterize the Amycolatopsis sp. AA4 - S. coelicolor interaction by examining expression of developmental and iron acquisition genes over time in co-culture. Manipulation of iron availability and/or growth near Amycolatopsis sp. AA4 led to alterations in expression of the critical developmental gene bldN, and other key downstream genes in the S. coelicolor transcriptional cascade. In Amycolatopsis sp. AA4, siderophore genes were downregulated when grown near S. coelicolor, leading us to find that deferrioxamine E, produced by S. coelicolor, could be readily utilized by Amycolatopsis sp. AA4. Collectively these results suggest that competition for iron via siderophore piracy and species-specific siderophores can alter patterns of gene expression and morphological differentiation during actinomycete interactions. PMID:22931126

  13. Glucose kinase has a regulatory role in carbon catabolite repression in Streptomyces coelicolor.

    PubMed Central

    Kwakman, J H; Postma, P W

    1994-01-01

    A glucose kinase (glkA) mutant of Streptomyces coelicolor A3(2) M145 was selected by the ability to grow in the presence of the nonmetabolizable glucose analog 2-deoxyglucose. In this glkA mutant, carbon catabolite repression of glycerol kinase and agarase was relieved on several carbon sources tested, even though most of these carbon sources are not metabolized via glucose kinase. This suggests that catabolite repression is not regulated by the flux through glucose kinase and that the protein itself has a regulatory role in carbon catabolite repression. A 10-fold overproduction of glucose kinase also results in relief of catabolite repression, suggesting that excess glucose kinase can titrate the repressing signal away. This could be achieved directly by competition of excess glucose kinase with its repressing form for binding sites on DNA promoter regions or indirectly by competition for binding of another regulatory protein. Images PMID:8169219

  14. Uncovering Genes with Divergent mRNA-Protein Dynamics in Streptomyces coelicolor

    PubMed Central

    Jayapal, Karthik P.; Philp, Robin J.; Kok, Yee-Jiun; Yap, Miranda G. S.; Sherman, David H.; Griffin, Timothy J.; Hu, Wei-Shou

    2008-01-01

    Many biological processes are intrinsically dynamic, incurring profound changes at both molecular and physiological levels. Systems analyses of such processes incorporating large-scale transcriptome or proteome profiling can be quite revealing. Although consistency between mRNA and proteins is often implicitly assumed in many studies, examples of divergent trends are frequently observed. Here, we present a comparative transcriptome and proteome analysis of growth and stationary phase adaptation in Streptomyces coelicolor, taking the time-dynamics of process into consideration. These processes are of immense interest in microbiology as they pertain to the physiological transformations eliciting biosynthesis of many naturally occurring therapeutic agents. A shotgun proteomics approach based on mass spectrometric analysis of isobaric stable isotope labeled peptides (iTRAQ™) enabled identification and rapid quantification of approximately 14% of the theoretical proteome of S. coelicolor. Independent principal component analyses of this and DNA microarray-derived transcriptome data revealed that the prominent patterns in both protein and mRNA domains are surprisingly well correlated. Despite this overall correlation, by employing a systematic concordance analysis, we estimated that over 30% of the analyzed genes likely exhibited significantly divergent patterns, of which nearly one-third displayed even opposing trends. Integrating this data with biological information, we discovered that certain groups of functionally related genes exhibit mRNA-protein discordance in a similar fashion. Our observations suggest that differences between mRNA and protein synthesis/degradation mechanisms are prominent in microbes while reaffirming the plausibility of such mechanisms acting in a concerted fashion at a protein complex or sub-pathway level. PMID:18461186

  15. Ribosylhopane, a novel bacterial hopanoid, as precursor of C35 bacteriohopanepolyols in Streptomyces coelicolor A3(2)

    PubMed Central

    Liu, Wenjun; Sakr, Elias; Schaeffer, Philippe; Talbot, Helen M.; Donisi, Janina; Härtner, Thomas; Kannenberg, Elmar; Takano, Eriko

    2014-01-01

    Streptomyces coelicolor A3(2) wild type produces aminobacteriohopanetriol as the only elongated C35 hopanoid. The hopanoid phenotype of two mutants bearing a deletion of genes from a previously identified hopanoid biosynthesis gene cluster provides clues on the formation of C35 bacteriohopanepolyols. orf14 encodes a putative nucleosidase: its deletion induces the accumulation of adenosylhopane that cannot be converted into ribosylhopane. orf18 encodes a putative transaminase: its deletion results in the accumulation of adenosylhopane, ribosylhopane and bacteriohopanetetrol. Ribosylhopane was hypothesized twenty years ago as a precursor for bacterial hopanoids but was never identified in a bacterium. Absence of the transaminase encoded by orf18 prevents the reductive amination of ribosylhopane into aminobacteriohopanetriol and induces its accumulation. Its reduction by an aldose reductase like enzyme produces bacteriohopanetetrol, which is normally not present in S. coelicolor. PMID:25155017

  16. Characterization of FabG and FabI of the Streptomyces coelicolor dissociated fatty acid synthase.

    PubMed

    Singh, Renu; Reynolds, Kevin A

    2015-03-01

    Streptomyces coelicolor produces fatty acids for both primary metabolism and for biosynthesis of the secondary metabolite undecylprodiginine. The first and last reductive steps during the chain elongation cycle of fatty acid biosynthesis are catalyzed by FabG and FabI. The S. coelicolor genome sequence has one fabI gene (SCO1814) and three likely fabG genes (SCO1815, SCO1345, and SCO1846). We report the expression, purification, and characterization of the corresponding gene products. Kinetic analyses revealed that all three FabGs and FabI are capable of utilizing both straight and branched-chain β-ketoacyl-NAC and enoyl-NAC substrates, respectively. Furthermore, only SCO1345 differentiates between ACPs from both biosynthetic pathways. The data presented provide the first experimental evidence that SCO1815, SCO1346, and SCO1814 have the catalytic capability to process intermediates in both fatty acid and undecylprodiginine biosynthesis. PMID:25662938

  17. Economic Development Projects and Jobs: Lessons from the Targeted Jobs Demonstration Program.

    ERIC Educational Resources Information Center

    Van Horn, Carl; And Others

    This guide, based on approaches for targeting jobs and business opportunities that were developed during the Targeted Jobs Demonstration Program (TJDP), contains strategies and techniques for ensuring that some of the benefits of economic development investments are directed to low-income individuals and small and minority businesses. Addressed in…

  18. In Search of the E. coli Compounds that Change the Antibiotic Production Pattern of Streptomyces coelicolor During Inter-species Interaction.

    PubMed

    Mavituna, Ferda; Luti, Khalid Jaber Kadhum; Gu, Lixing

    2016-08-01

    The aim of this work was to investigate the interaction between E.coli and Streptomyces coelicolor A3 (2) for the increased production of undecylprodigiosin and identify the E. coli actives mediating this inter-species interaction. The antibiotics of interest were the red-pigmented undecylprodigiosin and blue-pigmented actinorhodin. Pure cultures of S. coelicolor in a defined medium produced higher concentrations of actinorhodin compared to those of undecylprodigiosin. The latter however, is more important due to its immunosuppressive and antitumor properties. As a strategy to increase undecylprodigiosin production, we added separately, live cells and heat-killed cells of E. coli C600, and the cell-free supernatant of E. coli culture to S. coelicolor cultures in shake flasks. The interaction with live cells of E. coli altered the antibiotic production pattern and undecylprodigiosin production was enhanced by 3.5-fold compared to the pure cultures of S. coelicolor and actinorhodin decreased by 15-fold. The heat-killed cells of E. coli however, had no effect on antibiotic production. In all cases, growth and glucose consumption of S. coelicolor remained almost the same as those observed in the pure culture indicating that the changes in antibiotic production were not due to nutritional stress. Results with cell-free supernatant of E. coli culture indicated that the interaction between S. coelicolor and E. coli was mediated via diffusible molecule(s). Using a set of extraction procedures and agar-well diffusion bioassays, we isolated and preliminarily identified a class of compounds. For the preliminary verification, we added the compound which was the common chemical structural moiety in this class of compounds to the pure S. coelicolor cultures. We observed similar effects on antibiotic production as with the live E. coli cells and their supernatant indicating that this class of compounds secreted by E. coli indeed could act as actives during interspecies

  19. Freihoelser Forst Local Training Area Demonstration Project: Prescription development and installation

    SciTech Connect

    Hinchman, R.R.; Zellmer, S.D.; Severinghaus, W.D.; Brent, J.J.

    1989-04-01

    The Freiholser Forst Local Training Area (LTA) Rehabilitation Demonstration Project is part of the Integrated Training Area Management program being developed by the US Army Corps of Engineers` Construction Engineering Research Laboratory for the Seventh Army Training Command of the US Army in Europe. The rehabilitation demonstration project was begun in 1987 to develop and demonstrate rapid, cost-effective methods to stabilize the LTA`s barren, eroding maneuver areas and make training conditions more realistic. The sandy, infertile, and acidic soils at the LTA are considered the major factor limiting rehabilitation efforts there. The project involves the evaluation of three procedures to revegetate the soils, each incorporating identical methods for preparing the seedbed and a single seed mixture consisting of adapted, native species but using different soil amendments. All three treatments have satisfactorily reestablished vegetation and controlled erosion on the demonstration plots at the LTA, but their costs have varied widely.

  20. Freihoelser Forst Local Training Area Demonstration Project: Prescription development and installation

    SciTech Connect

    Hinchman, R.R.; Zellmer, S.D. . Energy Systems Div.); Severinghaus, W.D. ); Brent, J.J. )

    1989-04-01

    The Freiholser Forst Local Training Area (LTA) Rehabilitation Demonstration Project is part of the Integrated Training Area Management program being developed by the US Army Corps of Engineers' Construction Engineering Research Laboratory for the Seventh Army Training Command of the US Army in Europe. The rehabilitation demonstration project was begun in 1987 to develop and demonstrate rapid, cost-effective methods to stabilize the LTA's barren, eroding maneuver areas and make training conditions more realistic. The sandy, infertile, and acidic soils at the LTA are considered the major factor limiting rehabilitation efforts there. The project involves the evaluation of three procedures to revegetate the soils, each incorporating identical methods for preparing the seedbed and a single seed mixture consisting of adapted, native species but using different soil amendments. All three treatments have satisfactorily reestablished vegetation and controlled erosion on the demonstration plots at the LTA, but their costs have varied widely.

  1. Integrated gasification combined-cycle research development and demonstration activities in the US

    SciTech Connect

    Ness, H.M.; Brdar, R.D.

    1996-09-01

    The United States Department of Energy (DOE)`s Office of Fossil Energy, Morgantown Energy Technology Center, is managing a research development and demonstration (RD&D) program that supports the commercialization of integrated gasification combined-cycle (IGCC) advanced power systems. This overview briefly describes the supporting RD&D activities and the IGCC projects selected for demonstration in the Clean Coal Technology (CCT) Program.

  2. Integrated gasification combined-cycle research development and demonstration activities in the U.S.

    SciTech Connect

    Ness, H.M.

    1994-12-31

    The United States Department of Energy (DOE) has selected seven integrated gasification combined-cycle (IGCC) advanced power systems for demonstration in the Clean Coal Technology (CCT) Program. DOE`s Office of Fossil Energy, Morgantown Energy Technology Center, is managing a research development and demonstration (RD&D)program that supports the CCT program, and addresses long-term improvements in support of IGCC technology. This overview briefly describes the CCT projects and the supporting RD&D activities.

  3. Purification and characterization of recombinant malate synthase enzymes from Streptomyces coelicolor A3(2) and S. clavuligerus NRRL3585.

    PubMed

    Loke, P; Goh, L L; Seng Soh, B; Yeow, P; Sim, T S

    2002-04-01

    Malate synthases (MS) from Streptomyces coelicolor A3(2) and S. clavuligerus NRRL3585 were cloned by polymerase chain reaction into a glutathione S-transferase (GST) fusion expression vector and heterologously expressed in Escherichia coli. The fusion GST-MS construct improved the soluble expression of MS by approximately 10-fold compared to the soluble expression of nonfusion MS. With the significant improvement in levels of soluble MS, purification and subsequent cleavage of recombinant MS from GST were facilitated in this study. Using purified enzymes, optimized parameters, which achieved maximal specific activity, were established in the enzymatic assay for streptomycete MS. The average purified specific activities of S. coelicolor and S. clavuligerus MS were 26199 and 11821 nmol/mg min, respectively. Furthermore, enzymatic analysis revealed that the two streptomycete MS displayed a similar Km value for acetyl-CoA, but S. coelicolor MS had a Km value for glyoxylate that is approximately sixfold higher than S. clavuligerus MS. PMID:11986926

  4. Medium engineering for enhanced production of undecylprodigiosin antibiotic in Streptomyces coelicolor using oil palm biomass hydrolysate as a carbon source.

    PubMed

    Bhatia, Shashi Kant; Lee, Bo-Rahm; Sathiyanarayanan, Ganesan; Song, Hun-Seok; Kim, Junyoung; Jeon, Jong-Min; Kim, Jung-Ho; Park, Sung-Hee; Yu, Ju-Hyun; Park, Kyungmoon; Yang, Yung-Hun

    2016-10-01

    In this study, a biosugar obtained from empty fruit bunch (EFB) of oil palm by hot water treatment and subsequent enzymatic saccharification was used for undecylprodigiosin production, using Streptomyces coelicolor. Furfural is a major inhibitor present in EFB hydrolysate (EFBH), having a minimum inhibitory concentration (MIC) of 1.9mM, and it reduces utilization of glucose (27%), xylose (59%), inhibits mycelium formation, and affects antibiotic production. Interestingly, furfural was found to be a good activator of undecylprodigiosin production in S. coelicolor, which enhanced undecylprodigiosin production by up to 52%. Optimization by mixture analysis resulted in a synthetic medium containing glucose:furfural:ACN:DMSO (1%, 2mM, 0.2% and 0.3%, respectively). Finally, S. coelicolor was cultured in a fermenter in minimal medium with EFBH as a carbon source and addition of the components described above. This yielded 4.2μg/mgdcw undecylprodigiosin, which was 3.2-fold higher compared to that in un-optimized medium. PMID:26951741

  5. Development and Demonstration of a 25 Watt Thermophotovoltaic Power Source for a Hybrid Power System

    NASA Technical Reports Server (NTRS)

    Doyle, Edward; Shukla, Kailash; Metcalfe, Christopher

    2001-01-01

    The development of a propane-fueled, 25 W thermophotovoltaic (TPV) power source for use in a hybrid power system is described. The TPV power source uses a platinum emitting surface with an anti-reflective coating to radiate to gallium antimonide photocells, which converts the radiation to electric power. The development program started with the design and fabrication of an engineering prototype system. This was used as a component development vehicle to develop the technologies for the various components. A 25 W demonstration prototype was then designed and fabricated using the most advanced component approaches. The designs and test results from this development program are discussed.

  6. Waste separations and pretreatment technology development in the Underground Storage Tank--Integrated Demonstration Program

    SciTech Connect

    Cruse, J.M.

    1993-04-01

    The principal objective of the Underground Storage Tank Integrated Demonstration (UST-ID) Program is the demonstration and continued development of technologies suitable for the remediation of USTs. The most promising new technologies are selected for demonstration, testing, and evaluation. The objective is the eventual transfer of new technologies as a system to full-scale remediation at US Department of Energy (DOE) complexes and sites in the private sector. Technologies under development in the UST-ID Program are targeted toward use in remediation actions at the following five DOE participant sites: Hanford, Fernald, Idaho, Oak Ridge, and Savannah River. Combined, these participant sites have more than 300 USTs containing more than 100 m gal of high-level and low-level radioactive liquid waste. This paper focuses on the Waste Separations and Pretreatment area of the UST-ID, summarizing the currently funded technology development projects. In this area, several ``compact processing units`` (CPUs) are being developed to effect near-term demonstration and deployment of promising processes in actual tank environments. The CPU design and development approach is summarized. A number of programmatic issues are discussed, including onsite and offsite transportation of potentially contaminated processing systems, containment design criteria, including applicable codes and standards, and operation and maintenance of such systems in a developmental testing environment.

  7. X-37 Flight Demonstrator: A Building Block in NASA's Future Access to Space; X-37 Flight Demonstrator: Orbital Vehicle Technology Development Approach

    NASA Technical Reports Server (NTRS)

    Jacobson, David

    2004-01-01

    Project management issues and subsystems development for the X-37 flight demonstrator are covered in this viewgraph presentation. Subsystems profiled include: thermal protection systems, hot structures, and lithium-ion batteries.

  8. DEVELOPMENT AND DEMONSTRATION OF AN ULTRA LOW NOx COMBUSTOR FOR GAS TURBINES

    SciTech Connect

    NEIL K. MCDOUGALD

    2005-04-30

    Alzeta Corporation has developed surface-stabilized fuel injectors for use with lean premixed combustors which provide extended turndown and ultra-low NOX emission performance. These injectors use a patented technique to form interacting radiant and blue-flame zones immediately above a selectively-perforated porous metal surface. This allows stable operation at low reaction temperatures. This technology is being commercialized under the product name nanoSTAR. Initial tests demonstrated low NOX emissions but, were limited by flashback failure of the injectors. The weld seams required to form cylindrical injectors from flat sheet material were identified as the cause of the failures. The approach for this project was to first develop new fabrication methods to produce injectors without weld seams, verify similar emissions performance to the original flat sheet material and then develop products for microturbines and small gas turbines along parallel development paths. A 37 month project was completed to develop and test a surface stabilized combustion system for gas turbine applications. New fabrication techniques developed removed a technological barrier to the success of the product by elimination of conductive weld seams from the injector surface. The injectors demonstrated ultra low emissions in rig tests conducted under gas turbine operating conditions. The ability for injectors to share a common combustion chamber allowing for deployment in annular combustion liner was also demonstrated. Some further development is required to resolve integration issues related to specific engine constraints, but the nanoSTAR technology has clearly demonstrated its low emissions potential. The overall project conclusions can be summarized: (1) A wet-laid casting method successfully eliminated weld seams from the injector surface without degrading performance. (2) Gas turbine cycle analysis identified several injector designs and control schemes to start and load engines using

  9. DEVELOPMENT AND DEMONSTRATION OF POLYMER MICROENCAPSULATION OF MIXED WASTE USING KINETIC MIXER PROCESSING

    SciTech Connect

    LAGERAAEN,P.R.; KALB,P.D.; MILIAN,L.W.; ADAMS,J.W.

    1997-11-01

    Thermokinetic mixing was investigated as an alternative processing method for polyethylene microencapsulation, a technology well demonstrated for treatment of hazardous, low-level radioactive and low-level mixed wastes. Polyethylene encapsulation by extrusion has been previously shown to be applicable to a wide range of waste types but often pretreatment of the wastes is necessary due to process limitations regarding the maximum waste moisture content and particle size distribution. Development testing was conducted with kinetic mixing in order to demonstrate technology viability and show improved process applicability in these areas. Testing to establish process capabilities and relevant operating parameters was performed with waste surrogates including an aqueous evaporator concentrate and soil. Using a pilot-scale kinetic mixer which was installed and modified for this program, the maximum waste moisture content and particle size was determined. Following process development with surrogate wastes, the technology was successfully demonstrated at BNL using actual mixed waste.

  10. Research, development and demonstration of nickel-iron batteries for electric-vehicle propulsion

    NASA Astrophysics Data System (ADS)

    1983-05-01

    The development of nickel-iron batteries was addressed to demonstrate improved performance features in electric vehicle battery systems. Performance improvements reported include: 30% increase in gravimetric energy density to 57 Wh/kg, 70% increase in volumetric enegy density to 116 Wh/1, and 5% increase in power density to 105 W/kg. In addition, a preliminary design was established for a multicell modular package that can further enhance these performance parameters by providing weight and volume reductions. Additional accomplishments are reported in process development by operating pilot line facilities to demonstrate the potential for $80/kWh OEM selling price. The potential for meeting the cycle life goal was enhanced by demonstration of: iron electrode life of over 1000 cycles at 80% depth of discharge (DOD), nickel electrode life of over 1000 cycles at 80% DOD, cell life of over 1000 cycles at 80% DOD, and 5-cell module life of 903 cycles at 80% DOD.

  11. Development of Performance Demonstration Programs for Steam Generator Tubing Analysts in Korea

    SciTech Connect

    Chan-Hee, Cho; Min-Woo, Nam; Dong-Hyun, Jee; Jee-Hong, Jung; Hee-Jong, Lee; Se-Kyoung, Kim

    2006-07-01

    Korea Electric Power Research Institute developed the performance demonstration programs for non-destructive examination personnel who analyze eddy current data for steam generator tubes of the nuclear power plant. The purpose of these programs is to ensure a continuing uniform knowledge base and skill level for data analysts and contribute to safely operate the nuclear power plant. In Korea, there have been many changes for the non-destructive examination of steam generator tubing in the nuclear power plant such as inspection scope, plugging criteria and qualification requirements. According to the Notice 2004-13 revised by the Ministry of Science and Technology in Korea, the analysts for steam generator tubing shall be qualified as the Qualified Data Analyst, and the Site Specific Performance Demonstration program shall be implemented for them. KEPRI developed these performance demonstration programs and they are being successfully implemented. The analyst performance is expected to be improved by the implementation of theses programs. (authors)

  12. DEVELOPMENT AND DEMONSTRATION OF INDOOR RADON REDUCTION MEASURES FOR 10 HOMES IN CLINTON, NEW JERSEY

    EPA Science Inventory

    The report discusses the development and demonstration of indoor radon reduction methods for 10 houses in Clinton, New Jersey, where (in the spring of 1986) the New Jersey Department of Environmental Protection (DEP) located a cluster of houses with extremely high radon levels. T...

  13. INSTRUCTIONAL SYSTEMS DEVELOPMENT--A DEMONSTRATION AND EVALUATION PROJECT. FINAL REPORT.

    ERIC Educational Resources Information Center

    BARSON, JOHN

    FOUR UNIVERSITIES COOPERATED WITH THE INSTRUCTIONAL MEDIA CENTER AT MICHIGAN STATE UNIVERSITY TO TEST, DEMONSTRATE, AND REFINE A MODEL FOR MEDIA INNOVATION AND INSTRUCTIONAL DEVELOPMENT WHICH HAD BEEN DESIGNED IN A EARLIER PROJECT. MEDIA, EVALUATIVE, AND INSTRUCTIONAL SPECIALIST CAPABILITIES WERE TEAMED TO THE DECISION PROCESSES OF THE INSTRUCTORS…

  14. ςBldN, an Extracytoplasmic Function RNA Polymerase Sigma Factor Required for Aerial Mycelium Formation in Streptomyces coelicolor A3(2)

    PubMed Central

    Bibb, Maureen J.; Molle, Virginie; Buttner, Mark J.

    2000-01-01

    Sporulation mutants of Streptomyces coelicolor appear white because they are defective in the synthesis of the gray polyketide spore pigment, and such white (whi) mutants have been used to define 13 sporulation loci. whiN, one of five new whi loci identified in a recent screen of NTG (N-methyl-N′-nitro-N-nitrosoguanidine)-induced whi strains (N. J. Ryding et al., J. Bacteriol. 181:5419–5425, 1999), was defined by two mutants, R112 and R650. R650 produced frequent spores that were longer than those of the wild type. In contrast, R112 produced long, straight, undifferentiated hyphae, although rare spore chains were observed, sometimes showing highly irregular septum placement. Subcloning and sequencing showed that whiN encodes a member of the extracytoplasmic function subfamily of RNA polymerase sigma factors and that the sigma factor has an unusual N-terminal extension of approximately 86 residues that is not present in other sigma factors. A constructed whiN null mutant failed to form aerial mycelium (the “bald” phenotype) and, as a consequence, whiN was renamed bldN. This observation was not totally unexpected because, on some media, the R112 point mutant produced substantially less aerial mycelium than its parent, M145. The bldN null mutant did not fit simply into the extracellular signaling cascade proposed for S. coelicolor bld mutants. Expression of bldN was analyzed during colony development in wild-type and aerial mycelium-deficient bld strains. bldN was transcribed from a single promoter, bldNp. bldN transcription was developmentally regulated, commencing approximately at the time of aerial mycelium formation, and depended on bldG and bldH, but not on bldA, bldB, bldC, bldF, bldK, or bldJ or on bldN itself. Transcription from the p1 promoter of the response-regulator gene bldM depended on bldN in vivo, and the bldMp1 promoter was shown to be a direct biochemical target for ςBldN holoenzyme in vitro. PMID:10913095

  15. The phage growth limitation system in Streptomyces coelicolor A(3)2 is a toxin/antitoxin system, comprising enzymes with DNA methyltransferase, protein kinase and ATPase activity

    PubMed Central

    Hoskisson, Paul A.; Sumby, Paul; Smith, Margaret C.M.

    2015-01-01

    The phage growth limitation system of Streptomyces coelicolor A3(2) is an unusual bacteriophage defence mechanism. Progeny ϕC31 phage from an initial infection are thought to be modified such that subsequent infections are attenuated in a Pgl+ host but normal in a Pgl− strain. Earlier work identified four genes required for phage resistance by Pgl. Here we demonstrate that Pgl is an elaborate and novel phage restriction system that, in part, comprises a toxin/antitoxin system where PglX, a DNA methyltransferase is toxic in the absence of a functional PglZ. In addition, the ATPase activity of PglY and a protein kinase activity in PglW are shown to be essential for phage resistance by Pgl. We conclude that on infection of a Pgl+ cell by bacteriophage ϕC31, PglW transduces a signal, probably via phosphorylation, to other Pgl proteins resulting in the activation of the DNA methyltransferase, PglX and this leads to phage restriction. PMID:25592393

  16. Roles of two-component system AfsQ1/Q2 in regulating biosynthesis of the yellow-pigmented coelimycin P2 in Streptomyces coelicolor.

    PubMed

    Chen, Shuangshuang; Zheng, Guosong; Zhu, Hong; He, Huiqi; Chen, Lei; Zhang, Weiwen; Jiang, Weihong; Lu, Yinhua

    2016-08-01

    We previously demonstrated that in Streptomyces coelicolor two-component system AfsQ1/Q2 activates the production of the yellow-colored coelimycin P2 (also named as yCPK) on glutamate-supplemented minimal medium, and the response regulator AfsQ1 could specifically bind to the intergenic region between two structural genes, cpkA and cpkD Here, a more in-depth investigation was performed to elucidate the mechanism underlying the role of AfsQ1/Q2 in regulating coelimycin P2 biosynthesis. Deletion of afsQ1/Q2 resulted in markedly decreased expression of the whole coelimycin P2 biosynthetic gene cluster. Electrophoretic mobility shift assays revealed that AfsQ1 bound only to the target site identified previously, but not to any other promoters in the gene cluster. Mutations of AfsQ1-binding motif only resulted in drastically reduced transcription of the cpkA/B/C operon (encoding three type I polyketide synthases) and intriguingly, led to enhanced expression of some coelimcyin P2 genes, particularly accA1 and scF These results suggested the direct role of AfsQ1/Q2 in regulating coelimycin production, which is directly mediated by the structural genes, but not the cluster-situated regulatory genes, and also implied that other unknown mechanisms may be involved in AfsQ1/Q2-mediated regulation of coelimycin P2 biosynthesis. PMID:27313101

  17. Molecular characterization of SCO0765 as a cellotriose releasing endo-β-1,4-cellulase from Streptomyces coelicolor A(3).

    PubMed

    Hong, Joo-Bin; Dhakshnamoorthy, Vijayalakshmi; Lee, Chang-Ro

    2016-09-01

    The sco0765 gene was annotated as a glycosyl hydrolase family 5 endoglucanase from the genomic sequence of Streptomyces coelicolor A3(2) and consisted of 2,241 bp encoding a polypeptide of 747 amino acids (molecular weight of 80.5 kDa) with a 29-amino acid signal peptide for secretion. The SCO0765 recombinant protein was heterogeneously over-expressed in Streptomyces lividans TK24 under the control of a strong ermE* promoter. The purified SCO0765 protein showed the expected molecular weight of the mature form (718 aa, 77.6 kDa) on sodium dodecyl sulfate-polyacryl amide gel electrophoresis. SCO0765 showed high activity toward β-glucan and carboxymethyl cellulose (CMC) and negligible activity to Avicel, xylan, and xyloglucan. The SCO0765 cellulase had a maximum activity at pH 6.0 and 40°C toward CMC and at pH 9.0 and 50-60°C toward β-glucan. Thin layer chromatography of the hydrolyzed products of CMC and β-glucan by SCO0765 gave cellotriose as the major product and cellotetraose, cellopentaose, and longer oligosaccharides as the minor products. These results clearly demonstrate that SCO0765 is an endo-β-1,4-cellulase, hydrolyzing the β-1,4 glycosidic bond of cellulose into cellotriose. PMID:27572512

  18. Molecular and Functional Analyses of the Gene (eshA) Encoding the 52-Kilodalton Protein of Streptomyces coelicolor A3(2) Required for Antibiotic Production

    PubMed Central

    Kawamoto, Shinichi; Watanabe, Masakatsu; Saito, Natsumi; Hesketh, Andrew; Vachalova, Katerina; Matsubara, Keiko; Ochi, Kozo

    2001-01-01

    Analysis of proteins recovered in the S100 precipitate fraction of Streptomyces griseus after ultracentrifugation led to the identification of a 52-kDa protein which is produced during the late growth phase. The gene (eshA) which codes for this protein was cloned from S. griseus, and then its homologue was cloned from Streptomyces coelicolor A3(2). The protein was deduced to be 471 amino acids in length. The protein EshA is characterized by a central region that shows homology to the eukaryotic-type cyclic nucleotide-binding domains. Significant homology was also found to MMPI in Mycobacterium leprae, a major antigenic protein to humans. The eshA gene mapped near the chromosome end and was not essential for viability, as demonstrated by gene disruption experiments, but its disruption resulted in the abolishment of an antibiotic (actinorhodin but not undecylprodigiosin) production. Aerial mycelium was produced as abundantly as by the parent strain. Expression analysis of the EshA protein by Western blotting revealed that EshA is present only in late-growth-phase cells. The eshA gene was transcribed just preceding intracellular accumulation of the EshA protein, as determined by S1 nuclease protection, indicating that EshA expression is regulated at the transcription level. The expression of EshA was unaffected by introduction of the relA mutation, which blocks ppGpp synthesis. PMID:11567001

  19. Development of the Optical Communications Telescope Laboratory: A Laser Communications Relay Demonstration Ground Station

    NASA Technical Reports Server (NTRS)

    Wilson, K. E.; Antsos, D.; Roberts, L. C. Jr.,; Piazzolla, S.; Clare, L. P.; Croonquist, A. P.

    2012-01-01

    The Laser Communications Relay Demonstration (LCRD) project will demonstrate high bandwidth space to ground bi-directional optical communications links between a geosynchronous satellite and two LCRD optical ground stations located in the southwestern United States. The project plans to operate for two years with a possible extension to five. Objectives of the demonstration include the development of operational strategies to prototype optical link and relay services for the next generation tracking and data relay satellites. Key technologies to be demonstrated include adaptive optics to correct for clear air turbulence-induced wave front aberrations on the downlink, and advanced networking concepts for assured and automated data delivery. Expanded link availability will be demonstrated by supporting operations at small sun-Earth-probe angles. Planned optical modulation formats support future concepts of near-Earth satellite user services to a maximum of 1.244 Gb/s differential phase shift keying modulation and pulse position modulations formats for deep space links at data rates up to 311 Mb/s. Atmospheric monitoring instruments that will characterize the optical channel during the link include a sun photometer to measure atmospheric transmittance, a solar scintillometer, and a cloud camera to measure the line of sight cloud cover. This paper describes the planned development of the JPL optical ground station.

  20. Dual Positive Feedback Regulation of Protein Degradation of an Extra-cytoplasmic Function σ Factor for Cell Differentiation in Streptomyces coelicolor *

    PubMed Central

    Mao, Xu-Ming; Sun, Ning; Wang, Feng; Luo, Shuai; Zhou, Zhan; Feng, Wei-Hong; Huang, Fang-Liang; Li, Yong-Quan

    2013-01-01

    Here we report that in Streptomyces coelicolor, the protein stability of an ECF σ factor SigT, which is involved in the negative regulation of cell differentiation, was completely dependent on its cognate anti-σ factor RstA. The degradation of RstA caused a ClpP/SsrA-dependent degradation of SigT during cell differentiation. This was consistent with the delayed morphological development or secondary metabolism in the ΔclpP background after rstA deletion or sigT overexpression. Meanwhile, SigT negatively regulated clpP/ssrA expression by directly binding to the clpP promoter (clpPp). The SigT-clpPp interaction could be disrupted by secondary metabolites, giving rise to the stabilized SigT protein and retarded morphological development in a non-antibiotic-producing mutant. Thus a novel regulatory mechanism was revealed that the protein degradation of the ECF σ factor was initiated by the degradation of its anti-σ factor, and was accelerated in a dual positive feedback manner, through regulation by secondary metabolites, to promote rapid and irreversible development of the secondary metabolism. This ingenious cooperation of intracellular components can ensure economical and exquisite control of the ECF σ factor protein level for the proper cell differentiation in Streptomyces. PMID:24014034

  1. TRL Assessment of Solar Sail Technology Development Following the 20-Meter System Ground Demonstrator Hardware Testing

    NASA Technical Reports Server (NTRS)

    Young, Roy M.; Adams, Charles L.

    2010-01-01

    The NASA In-Space Propulsion Technology (ISPT) Projects Office sponsored two separate, independent solar sail system design and development demonstration activities during 2002-2005. ATK Space Systems of Goleta, CA was the prime contractor for one development team and L' Garde, Inc. of Tustin, CA was the prime contractor for the other development team. The goal of these activities was to advance the technology readiness level (TRL) of solar sail propulsion from 3 towards 6 by the year 2006. Component and subsystem fabrication and testing were completed successfully, including the ground deployment of 10-meter and 20-meter demonstration hardware systems under vacuum conditions. The deployment and structural testing of the 20-meter solar sail systems was conducted in the 30 meter diameter Space Power Facility thermal-vacuum chamber at NASA Glenn Plum Brook in April though August, 2005. This paper will present the results of the TRL assessment following the solar sail technology development activities associated with the design, development, analysis and testing of the 20-meter system ground demonstrators.

  2. Waste-heat research, development, demonstration and commercialization plan: Rankine-cycle bottoming systems. Executive summary

    SciTech Connect

    Not Available

    1980-01-01

    Organic and binary Rankine cycle (ORC) technology as potentially broad applications in recovering and converting waste heat to the useful energy form of electricity. ORC systems are particularly suited for recovering medium-grade exhaust heat (200 to 1000/sup 0/F), a form of waste energy released primarily in the generation of electricity and in industrial processes. Therefore, a Waste Heat Research, Development, Demonstration and Commercialization Plan (RDD and C Plan) has been formulated. The objective of the plan is to achieve significant market penetration of ORC technology by 1985. To accomplish this commercialization objective, the plan is structured around three key strategic elements: demonstration of technically and economically attractive ORC systems that meet the specific needs of commercial waste heat markets; stimulation of the demand for ORC products in these markets; and promotion of the development of a competitive industry to serve ORC markets efficiently. The development of this plan and ERDA's role in its implementation are discussed.

  3. Aging aircraft NDI Development and Demonstration Center (AANC): An overview. [nondestructive inspection

    NASA Technical Reports Server (NTRS)

    Walter, Patrick L.

    1992-01-01

    A major center with emphasis on validation of nondestructive inspection (NDI) techniques for aging aircraft, the Aging Aircraft NDI Development and Demonstration Center (AANC), has been funded by the FAA at Sandia National Laboratories. The Center has been assigned specific tasks in developing techniques for the nondestructive inspection of static engine parts, assessing inspection reliability (POD experiments), developing testbeds for NDI validation, maintaining a FAA library of characterized aircraft structural test specimens, and leasing a hangar to house a high flight cycle transport aircraft for use as a full scale test bed.

  4. First demonstration of electron scattering using a novel target developed for short-lived nuclei.

    PubMed

    Suda, T; Wakasugi, M; Emoto, T; Ishii, K; Ito, S; Kurita, K; Kuwajima, A; Noda, A; Shirai, T; Tamae, T; Tongu, H; Wang, S; Yano, Y

    2009-03-13

    We carried out a demonstrative electron scattering experiment using a novel ion-trap target exclusively developed for short-lived highly unstable nuclei. Using stable 133Cs ion as a target, this experiment completely mimicked electron scattering off short-lived nuclei. Achieving a luminosity higher than 10;{26} cm;{-2} s;{-1} with around only 10;{6} trapped ions on the electron beam, the angular distribution of elastic scattering was successfully measured. This experiment clearly demonstrates that electron scattering off rarely produced short-lived nuclei is practical with this target technique. PMID:19392108

  5. First Demonstration of Electron Scattering Using a Novel Target Developed for Short-Lived Nuclei

    SciTech Connect

    Suda, T.; Wakasugi, M.; Emoto, T.; Ito, S.; Wang, S.; Yano, Y.; Ishii, K.; Kurita, K.; Kuwajima, A.; Tamae, T.; Noda, A.; Shirai, T.; Tongu, H.

    2009-03-13

    We carried out a demonstrative electron scattering experiment using a novel ion-trap target exclusively developed for short-lived highly unstable nuclei. Using stable {sup 133}Cs ion as a target, this experiment completely mimicked electron scattering off short-lived nuclei. Achieving a luminosity higher than 10{sup 26} cm{sup -2} s{sup -1} with around only 10{sup 6} trapped ions on the electron beam, the angular distribution of elastic scattering was successfully measured. This experiment clearly demonstrates that electron scattering off rarely produced short-lived nuclei is practical with this target technique.

  6. The Formative Process Used by the Satellite Technology Demonstration in the Development of Television Programming for Junior High School Students. Satellite Technology Demonstration, Technical Report No. 0210.

    ERIC Educational Resources Information Center

    Connolly, A. J.; And Others

    The Satellite Technology Demonstration (STD) created a planning or formative process in the creation of an educational television series. The STD recommendations were: (1) define the process, in-depth, before developing the process; (2) make certain the process can adapt to unforeseen developments; (3) test the process by converting early scripts…

  7. Design, Development, And Testing of Umbilical System Mechanisms for the X-33 Advanced Technology Demonstrator

    NASA Technical Reports Server (NTRS)

    Littlefield, Alan C.; Melton, Gregory S.

    2000-01-01

    The X-33 Advanced Technology Demonstrator is an un-piloted, vertical take-off, horizontal landing spacecraft. The purpose of the X-33 program is to demonstrate technologies that will dramatically lower the cost of access to space. The rocket-powered X-33 will reach an altitude of up to 100 km and speeds between Mach 13 and 15. Fifteen flight tests are planned, beginning in 2000. Some of the key technologies demonstrated will be the linear aerospike engine, improved thermal protection systems, composite fuel tanks and reduced operational timelines. The X-33 vehicle umbilical connections provide monitoring, power, cooling, purge, and fueling capability during horizontal processing and vertical launch operations. Two "rise-off" umbilicals for the X-33 have been developed, tested, and installed. The X-33 umbilical systems mechanisms incorporate several unique design features to simplify horizontal operations and provide reliable disconnect during launch.

  8. Development of a 15 T $Nb_3Sn$ Accelerator Dipole Demonstrator at Fermilab

    SciTech Connect

    Novitski, I.; Andreev, N.; Barzi, E.; Carmichael, J.; Kashikhin, V. V.; Turrion, D.; Yu, M.; Zlobin, A. V.

    2015-01-01

    100 TeV scale Hadron Collider (HC) with a nominal operation field of at least 15 T is being considered for the post-LHC era, which requires using the $Nb_3Sn$ technology. Practical demonstration of this field level in an accelerator-quality magnet and substantial reduction of the magnet costs are the key conditions for realization of such a machine. FNAL has started the development of a 15 T Nb3Sn dipole demonstrator for a 100 TeV scale HC. The magnet design is based on 4-layer shell type coils, graded between the inner and outer layers to maximize the performance and reduce the cost. The experience gained during the Nb3Sn magnet R&D is applied to different aspects of the magnet design. This paper describes the magnetic and structural designs and parameters of the 15 T Nb3Sn dipole and the steps towards the demonstration model fabrication.

  9. Design, Development,and Testing of Umbillical System Mechanisms for the X-33 Advanced Technology Demonstrator

    NASA Technical Reports Server (NTRS)

    Littlefield, Alan C.; Melton, Gregory S.

    1999-01-01

    The X-33 Advanced Technology Demonstrator is an un-piloted, vertical take-off, horizontal landing spacecraft. The purpose of the X-33 program is to demonstrate technologies that will dramatically lower the cost of access to space. The rocket-powered X-33 will reach an altitude of up to 100 km and speeds between Mach 13 and 15. Fifteen flight tests are planned, beginning in 2000. Some of the key technologies demonstrated will be the linear aerospike engine, improved thermal protection systems, composite fuel tanks and reduced operational timelines. The X-33 vehicle umbilical connections provide monitoring, power, cooling, purge, and fueling capability during horizontal processing and vertical launch operations. Two "rise-ofF' umbilicals for the X-33 have been developed, tested, and installed. The X-33 umbilical systems mechanisms incorporate several unique design features to simplify horizontal operations and provide reliable disconnect during launch.

  10. Mixed Waste Focus Area alternative oxidation technologies development and demonstration program

    SciTech Connect

    Borduin, L.C.; Fewell, T.; Gombert, D.; Priebe, S.

    1998-07-01

    The Mixed Waste Focus Area (MWFA) is currently supporting the development and demonstration of several alternative oxidation technology (AOT) processes for treatment of combustible mixed low-level wastes. The impetus for this support derives from regulatory and political hurdles frequently encountered by traditional thermal techniques, primarily incinerators. AOTs have been defined as technologies that destroy organic material without using open-flame reactions. Whether thermal or nonthermal, the processes have the potential advantages of relatively low-volume gaseous emissions, generation of few or no dioxin/furan compounds, and operation at low enough temperatures that metals (except mercury) and most radionuclides are not volatilized. Technology development and demonstration are needed to confirm and realize the potential of AOTs and to compare them on an equal basis with their fully demonstrated thermal counterparts. AOTs include both thermal and nonthermal processes that oxidize organic wastes but operate under significantly different physical and chemical conditions than incinerators. Nonthermal processes currently being studied include Delphi DETOX and acid digestion at the Savannah River Site, and direct chemical oxidation at Lawrence Livermore National Laboratory. All three technologies are at advanced stages of development or are entering the demonstration phase. Nonflame thermal processes include catalytic chemical oxidation, which is being developed and deployed at Lawrence Berkeley National Laboratory, and team reforming, a commercial process being supported by Department of Energy. Related technologies include two low-flow, secondary oxidation processes (Phoenix and Thermatrix units) that have been tested at MSE, Inc., in Butte, Montana. Although testing is complete on some AOT technologies, most require additional support to complete some or all of the identified development objectives. Brief descriptions, status, and planned paths forward for each

  11. The Iodine Satellite (iSAT) Hall Thruster Demonstration Mission Concept and Development

    NASA Technical Reports Server (NTRS)

    Dankanich, John W.; Polzin, Kurt A.; Calvert, Derek; Kamhawi, Hani

    2014-01-01

    The use of iodine propellant for Hall thrusters has been studied and proposed by multiple organizations due to the potential mission benefits over xenon. In 2013, NASA Marshall Space Flight Center competitively selected a project for the maturation of an iodine flight operational feed system through the Technology Investment Program. Multiple partnerships and collaborations have allowed the team to expand the scope to include additional mission concept development and risk reduction to support a flight system demonstration, the iodine Satellite (iSAT). The iSAT project was initiated and is progressing towards a technology demonstration mission preliminary design review. The current status of the mission concept development and risk reduction efforts in support of this project is presented.

  12. Crew station research and development facility training for the light helicopter demonstration/validation program

    NASA Technical Reports Server (NTRS)

    Matsumoto, Joy Hamerman; Rogers, Steven; Mccauley, Michael; Salinas, AL

    1992-01-01

    The U.S. Army Crew Station Research and Development Branch (CSRDB) of the Aircraft Simulation Division (AVSCOM) was tasked by the Light Helicopter Program Manager (LH-PM) to provide training to Army personnel in advanced aircraft simulation technology. The purpose of this training was to prepare different groups of pilots to support and evaluate two contractor simulation efforts during the Demonstration/Validation (DEM/VAL) phase of the LH program. The personnel in the CSRDB developed mission oriented training programs to accomplish the objectives, conduct the programs, and provide guidance to army personnel and support personnel throughout the DEM/VAL phase.

  13. Vibration isolation technology: An executive summary of systems development and demonstration

    NASA Technical Reports Server (NTRS)

    Grodsinsky, Carlos M.; Logsdon, Kirk A.; Lubomski, Joseph F.

    1993-01-01

    A program was organized to develop the enabling technologies needed for the use of Space Station Freedom as a viable microgravity experimental platform. One of these development programs was the Vibration Isolation Technology (VIT). This technology development program grew because of increased awareness that the acceleration disturbances present on the Space Transportation System (STS) orbiter can and are detrimental to many microgravity experiments proposed for STS, and in the future, Space Station Freedom (SSF). Overall technological organization are covered of the VIT program. Emphasis is given to the results from development and demonstration of enabling technologies to achieve the acceleration requirements perceived as those most likely needed for a variety of microgravity science experiments. In so doing, a brief summary of general theoretical approaches to controlling the acceleration environment of an isolated space based payload and the design and/or performance of two prototype six degree of freedom active magnetic isolation systems is presented.

  14. 1000kW on-site PAFC power plant development and demonstration

    SciTech Connect

    Satomi, Tomohide; Koike, Shunichi; Ishikawa, Ryou

    1996-12-31

    Phosphoric Acid Fuel Cell Technology Research Association (PAFC-TRA) and New Energy and Industrial Technology Development Organization (NEDO) have been conducting a joint project on development of a 5000kW urban energy center type PAFC power plant (pressurized) and a 1000kW on-site PAFC power plant (non-pressurized). The objective of the technical development of 1000kW on-site PAFC power plant is to realize a medium size power plant with an overall efficiency of over 70% and an electrical efficiency of over 36%, that could be installed in a large building as a cogeneration system. The components and system integration development work and the plant design were performed in 1991 and 1992. Manufacturing of the plant and installation at the test site were completed in 1994. PAC test was carried out in 1994, and generation test was started in January 1995. Demonstration test is scheduled for 1995 and 1996.

  15. Microgravity vibration isolation technology: Development to demonstration. Ph.D. Thesis - Case Western Reserve Univ.

    NASA Technical Reports Server (NTRS)

    Grodsinsky, Carlos M.

    1993-01-01

    The low gravity environment provided by space flight has afforded the science community a unique area for the study of fundamental and technological sciences. However, the dynamic environment observed on space shuttle flights and predicted for Space Station Freedom has complicated the analysis of prior 'microgravity' experiments and prompted concern for the viability of proposed space experiments requiring long term, low gravity environments. Thus, isolation systems capable of providing significant improvements to this random environment have been developed. This dissertation deals with the design constraints imposed by acceleration sensitive, microgravity experiment payloads in the unique environment of space. A theoretical background for the inertial feedback and feedforward isolation of a payload was developed giving the basis for two experimental active inertial isolation systems developed for the demonstration of these advanced active isolation techniques. A prototype six degree of freedom digital active isolation system was designed and developed for the ground based testing of an actively isolated payload in three horizontal degrees of freedom. A second functionally equivalent system was built for the multi-dimensional testing of an active inertial isolation system in a reduced gravity environment during low gravity aircraft trajectories. These multi-input multi-output control systems are discussed in detail with estimates on acceleration noise floor performance as well as the actual performance acceleration data. The attenuation performance is also given for both systems demonstrating the advantages between inertial and non-inertial control of a payload for both the ground base environment and the low gravity aircraft acceleration environment. A future goal for this area of research is to validate the technical approaches developed to the 0.01 Hz regime by demonstrating a functional active inertial feedforward/feedback isolation system during orbital flight

  16. TRL Assessment of Solar Sail Technology Development Following the 20-Meter System Ground Demonstrator Hardware Testing

    NASA Technical Reports Server (NTRS)

    Young, Roy M.; Montgomery, Edward E.; Montgomery, Sandy; Adams, Charles L.

    2007-01-01

    The NASA In-Space Propulsion Technology (ISPT) Projects Office has been sponsoring 2 separate, independent system design and development hardware demonstration activities during 2002-2005. ATK Space Systems of Goleta, CA was the prime contractor for one development team and L'Garde, Inc. of Tustin, CA was the prime contractor for the other development team. The goal of these activities was to advance the technology readiness level (TRL) of solar sail propulsion from 3 towards 6 by the year 2006. Component and subsystem fabrication and testing were completed successfully, including the ground deployment of 10-meter and 20-meter ground demonstration hardware systems under vacuum conditions. The deployment and structural testing of the 20-meter solar sail systems was conducted in the 30 meter diameter Space Power Facility thermal-vacuum chamber at NASA Glenn Plum Brook in April though August, 2005. This paper will present the results of the TRL assessment following the solar sail technology development activities associated with the design, development, analysis and testing of the 20-meter system ground demonstrators. Descriptions of the system designs for both the ATK and L'Garde systems will be presented. Changes, additions and evolution of the system designs will be highlighted. A description of the modeling and analyses activities performed by both teams, as well as testing conducted to raise the TRL of solar sail technology will be presented. A summary of the results of model correlation activities will be presented. Finally, technology gaps identified during the assessment and gap closure plans will be presented, along with "lessons learned", subsequent planning activities and validation flight opportunities for solar sail propulsion technology.

  17. Highly efficient editing of the actinorhodin polyketide chain length factor gene in Streptomyces coelicolor M145 using CRISPR/Cas9-CodA(sm) combined system.

    PubMed

    Zeng, Hu; Wen, Shishi; Xu, Wei; He, Zhaoren; Zhai, Guifa; Liu, Yunkun; Deng, Zixin; Sun, Yuhui

    2015-12-01

    The current diminishing returns in finding useful antibiotics and the occurrence of drug-resistant bacteria call for the need to find new antibiotics. Moreover, the whole genome sequencing revealed that the biosynthetic potential of Streptomyces, which has produced the highest numbers of approved and clinical-trial drugs, has been greatly underestimated. Considering the known gene editing toolkits were arduous and inefficient, novel and efficient gene editing system are desirable. Here, we developed an engineered CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein) combined with the counterselection system CodA(sm), the D314A mutant of cytosine deaminase, to rapidly and effectively edit Streptomyces genomes. In-frame deletion of the actinorhodin polyketide chain length factor gene actI-ORF2 was created in Streptomyces coelicolor M145 as an illustration. This CRISPR/Cas9-CodA(sm) combined system strikingly increased the frequency of unmarked mutants and shortened the time required to generate them. We foresee the system becoming a routine laboratory technique for genome editing to exploit the great biosynthetic potential of Streptomyces and perhaps for other medically and economically important actinomycetes. PMID:26318449

  18. Highly efficient synthesis of chiral alcohols with a novel NADH-dependent reductase from Streptomyces coelicolor.

    PubMed

    Wang, Li-Juan; Li, Chun-Xiu; Ni, Yan; Zhang, Jie; Liu, Xiang; Xu, Jian-He

    2011-07-01

    An NADH-dependent reductase (ScCR) from Streptomyces coelicolor was discovered by genome mining for carbonyl reductases. ScCR was overexpressed in Escherichia coli BL21, purified to homogeneity and its catalytic properties were studied. This enzyme catalyzed the asymmetric reduction of a broad range of prochiral ketones including aryl ketones, α- and β-ketoesters, with high activity and excellent enantioselectivity (>99% ee) towards β-ketoesters. Among them, ethyl 4-chloro-3-oxobutanoate (COBE) was efficiently converted to ethyl (S)-4-chloro-3-hydroxybutanoate ((S)-CHBE), an important pharmaceutical intermediate, in water/toluene biphasic system. As much as 600 g/L (3.6M) of COBE was asymmetrically reduced within 22 h using 2-propanol as a co-substrate for NADH regeneration, resulting in a yield of 93%, an enantioselectivity of >99% ee, and a total turnover number (TTN) of 12,100. These results indicate the potential of ScCR for the industrial production of valuable chiral alcohols. PMID:21570826

  19. Complex Intra-Operonic Dynamics Mediated by a Small RNA in Streptomyces coelicolor

    PubMed Central

    Hindra; Moody, Matthew J.; Jones, Stephanie E.; Elliot, Marie A.

    2014-01-01

    Streptomyces are predominantly soil-dwelling bacteria that are best known for their multicellular life cycle and their prodigious metabolic capabilities. They are also renowned for their regulatory capacity and flexibility, with each species encoding >60 sigma factors, a multitude of transcription factors, and an increasing number of small regulatory RNAs. Here, we describe our characterization of a conserved small RNA (sRNA), scr4677. In the model species Streptomyces coelicolor, this sRNA is located in the intergenic region separating SCO4677 (an anti-sigma factor-encoding gene) and SCO4676 (a putative regulatory protein-encoding gene), close to the SCO4676 translation start site in an antisense orientation. There appears to be considerable genetic interplay between these different gene products, with wild type expression of scr4677 requiring function of the anti-sigma factor SCO4677, and scr4677 in turn influencing the abundance of SCO4676-associated transcripts. The scr4677-mediated effects were independent of RNase III (a double stranded RNA-specific nuclease), with RNase III having an unexpectedly positive influence on the level of SCO4676-associated transcripts. We have shown that both SCO4676 and SCO4677 affect the production of the blue-pigmented antibiotic actinorhodin under specific growth conditions, and that this activity appears to be independent of scr4677. PMID:24465751

  20. AllR Controls the Expression of Streptomyces coelicolor Allantoin Pathway Genes

    PubMed Central

    Navone, Laura; Macagno, Juan Pablo; Licona-Cassani, Cuauhtémoc; Marcellin, Esteban; Nielsen, Lars K.; Gramajo, Hugo

    2015-01-01

    Streptomyces species are native inhabitants of soil, a natural environment where nutrients can be scarce and competition fierce. They have evolved ways to metabolize unusual nutrients, such as purines and its derivatives, which are highly abundant in soil. Catabolism of these uncommon carbon and nitrogen sources needs to be tightly regulated in response to nutrient availability and environmental stimulus. Recently, the allantoin degradation pathway was characterized in Streptomyces coelicolor. However, there are questions that remained unanswered, particularly regarding pathway regulation. Here, using a combination of proteomics and genetic approaches, we identified the negative regulator of the allantoin pathway, AllR. In vitro studies confirmed that AllR binds to the promoter regions of allantoin catabolic genes and determined the AllR DNA binding motif. In addition, effector studies showed that allantoic acid, and glyoxylate, to a lesser extent, inhibit the binding of AllR to the DNA. Inactivation of AllR repressor leads to the constitutive expression of the AllR regulated genes and intriguingly impairs actinorhodin and undecylprodigiosin production. Genetics and proteomics analysis revealed that among all genes from the allantoin pathway that are upregulated in the allR mutant, the hyi gene encoding a hydroxypyruvate isomerase (Hyi) is responsible of the impairment of antibiotic production. PMID:26187964

  1. Conditional control of gene expression by synthetic riboswitches in Streptomyces coelicolor.

    PubMed

    Rudolph, Martin M; Vockenhuber, Michael-Paul; Suess, Beatrix

    2015-01-01

    Here we provide a step-by-step protocol for the application of synthetic theophylline-dependent riboswitches for conditional gene expression in Streptomyces coelicolor. Application of the method requires a sequence of only ~85 nt to be inserted between the transcriptional start site and the start codon of a gene of interest. No auxiliary factors are needed. All tested riboswitch variants worked well in concert with the promoters galP2, ermEp1, and SF14. Moreover, they allowed theophylline-dependent expression not only of the heterologous β-glucuronidase reporter gene but also of dagA, an endogenous agarase gene. The right combination of the tested promoters with the riboswitch variants allows for the adjustment of the desired dynamic range of regulation in a highly specific and dose-dependent manner and underlines the orthogonality of riboswitch regulation. We anticipate that any additional natural or synthetic promoter can be combined with the presented riboswitches. Moreover, this system should easily be transferable to other Streptomyces species, and most likely to any other genetically manipulable bacteria. PMID:25605391

  2. FY94 Office of Technology Development Mixed Waste Operations Robotics Demonstration

    SciTech Connect

    Kriikku, E.M.

    1994-08-30

    The Department of Energy (DOE) Office of Technology Development (OTD) develops technologies to help solve waste management and environmental problems at DOE sites. The OTD includes the Robotics Technology Development Program (RTDP) and the Mixed Waste Integrated Program (MWIP). Together these programs will provide technologies for DOE mixed waste cleanup projects. Mixed waste contains both radioactive and hazardous constituents. DOE sites currently store over 240,000 cubic meters of low level mixed waste and cleanup activities will generate several hundred thousand more cubic meters. Federal and state regulations require that this waste must be processed before final disposal. The OTD RTDP Mixed Waste Operations (MWO) team held several robotic demonstrations at the Savannah River Site (SRS) during November of 1993. Over 330 representatives from DOE, Government Contractors, industry, and universities attended. The MWO team includes: Fernald Environmental Management Project (FEMP), Idaho National Engineering Laboratory (INEL), Lawrence Livermore National Laboratory (LLNL), Oak Ridge National Engineering Laboratory (ORNL), Sandia National Laboratory (SNL), and Savannah River Technology Center (SRTC). SRTC is the lead site for MWO and provides the technical coordinator. The primary demonstration objective was to show that robotic technologies can make DOE waste facilities run better, faster, more cost effective, and safer. To meet the primary objective, the demonstrations successfully showed the following remote waste drum processing activities: non-destructive drum examination, drum transportation, drum opening, removing waste from a drum, characterize and sort waste items, scarify metal waste, and inspect stored drums. To further meet the primary objective, the demonstrations successfully showed the following remote waste box processing activities: swing free crane control, workcell modeling, and torch standoff control.

  3. Development and demonstration of solvent extraction processes for the separation of radionuclides from acidic radioactive waste

    SciTech Connect

    Law, J.D.; Brewer, K.N.; Herbst, R.S.; Todd, T.A.; Wood, D.J.

    1999-06-01

    The presence of long-lived radionuclides presents a challenge to the management of radioactive wastes. Immobilization of these radionuclides must be accomplished prior to long-term, permanent disposal. Separation of the radionuclides from the waste solutions has the potential of significantly decreasing the costs associated with the immobilization and disposal of the radioactive waste by minimizing waste volumes. Several solvent extraction processes have been developed and demonstrated at the Idaho National Engineering and Environmental Laboratory for the separation of transuranic element (TRUs), {sup 90}Sr, and/or {sup 137}Cs from acidic radioactive waste solutions. The Transuranic Extraction (TRUEX) and phosphine oxide (POR) processes for the separation of TRUs, the Strontium Extraction (SREX) process for the separation of {sup 90}Sr, the chlorinated cobalt dicarbollide (ChCoDiC) process for the separation of {sup 137}Cs and {sup 90}Sr, and a universal solvent extraction process for the simultaneous separation of TRUs, {sup 90}Sr, and {sup 137}Cs have all been demonstrated in centrifugal contactors using actual radioactive waste solutions. This article summarizes the most recent results of each of the flowsheet demonstrations and allows for comparison of the technologies. The successful demonstration of these solvent extraction processes indicates that they are all viable for the treatment of acidic radioactive waste solutions.

  4. Affordable Development and Demonstration of a Small NTR Engine and Stage: How Small is Big Enough?

    NASA Technical Reports Server (NTRS)

    Borowski, S. K.; Sefcik, R. J.; Fittje, J. E.; McCurdy, D. R.; Qualls, A. L.; Schnitzler, B. G.; Werner, J.; Weitzberg, A.; Joyner, C. R.

    2015-01-01

    In FY11, NASA formulated a plan for Nuclear Thermal Propulsion (NTP) development that included Foundational Technology Development followed by system-level Technology Demonstrations The ongoing NTP project, funded by NASAs Advanced Exploration Systems (AES) program, is focused on Foundational Technology Development and includes 5 key task activities:(1) Fuel element fabrication and non-nuclear validation testing of heritage fuel options;(2) Engine conceptual design;(3) Mission analysis and engine requirements definition;(4) Identification of affordable options for ground testing; and(5) Formulation of an affordable and sustainable NTP development program Performance parameters for Point of Departure designs for a small criticality-limited and full size 25 klbf-class engine were developed during FYs 13-14 using heritage fuel element designs for both RoverNERVA Graphite Composite (GC) and Ceramic Metal (Cermet) fuel forms To focus the fuel development effort and maximize use of its resources, the AES program decided, in FY14, that a leader-follower down selection between GC and cermet fuel was required An Independent Review Panel (IRP) was convened by NASA and tasked with reviewing the available fuel data and making a recommendation to NASA. In February 2015, the IRP recommended and the AES program endorsed GC as the leader fuel In FY14, a preliminary development schedule DDTE plan was produced by GRC, DOE industry for the AES program. Assumptions, considerations and key task activities are presented here Two small (7.5 and 16.5 klbf) engine sizes were considered for ground and flight technology demonstration within a 10-year timeframe; their ability to support future human exploration missions was also examined and a recommendation on a preferred size is provided.

  5. Development and Demonstration of Sustainable Surface Infrastructure for Moon/Mars Exploration

    NASA Technical Reports Server (NTRS)

    Sanders, Gerald B.; Larson, William E.; Picard, Martin

    2011-01-01

    For long-term human exploration of the Moon and Mars to be practical, affordable, and sustainable, future missions must be able to identify and utilize resources at the site of exploration. The ability to characterize, extract, processes, and separate products from local material, known as In-Situ Resource Utilization (ISRU), can provide significant reductions in launch mass, logistics, and development costs while reducing risk through increased mission flexibility and protection as well as increased mission capabilities in the areas of power and transportation. Making mission critical consumables like propellants, fuel cell reagents and life support gases, as well as in-situ crew/hardware protection and energy storage capabilities can significantly enhance robotic and human science and exploration missions, however other mission systems need to be designed to interface with and utilize these in-situ developed products and services from the start or the benefits will be minimized or eliminated. This requires a level of surface and transportation system development coordination not typically utilized during early technology and system development activities. An approach being utilized by the US National Aeronautics and Space Administration and the Canadian Space Agency has been to utilize joint analogue field demonstrations to focus technology development activities to demonstrate and integrate new and potentially game changing. mission critical capabilities that would enable an affordable and sustainable surface infrastructure for lunar and Mars robotic and human exploration. Two analogue field tests performed in November 2008 and February 2010 demonstrated first generation capabilities for lunar resource prospecting, exploration site preparation, and oxygen extraction from regolith while initiating integration with mobility, science, fuel cell power, and propulsion disciplines. A third analogue field test currently planned for June 2012 will continue and expand

  6. Technical support to the Solvent Refined Coal (SRC) demonstration projects: assessment of current research and development

    SciTech Connect

    Edwards, M.S.; Rodgers, B.R.; Brown, C.H.; Carlson, P.K.; Gambill, W.R.; Gilliam, T.M.; Holmes, J.M.; Krishnan, R.P.; Parsly, L.F.

    1980-12-01

    A program to demonstrate Solvent Refined Coal (SRC) technology has been initiated by the US Department of Energy (DOE) in partnership with two industrial groups. Project management responsibility has been assigned to the Oak Ridge Operations Office (ORO) of DOE. ORO requested that the Oak Ridge National Laboratory assess current research and development (R and D) activities and develop recommendations for those activities that might contribute to successful completion of the SRC demonstration plant projects. The objectives of this final report are to discuss in detail the problem areas in SRC; to discuss the current and planned R and D investigations relevant to the problems identified; and to suggest appropriate R and D activities in support of designs for the SRC demonstration plants. Four types of R and D activities are suggested: continuation of present and planned activities; coordination of activities and results, present and proposed; extension/redirection of activities not involving major equipment purchase or modifications; and new activities. Important examples of the first type of activity include continuation of fired heater, slurry rheology, and slurry mixing studies at Ft. Lewis. Among the second type of activity, coordination of data acquisition and interpretation is recommended in the areas of heat transfer, vapor/liquid equilibria, and physical properties. Principal examples of recommendations for extension/redirection include screening studies at laboratory scale on the use of carbonaceous precoat (e.g., anthracite) infiltration, and 15- to 30-day continuous tests of the Texaco gasifier at the Texaco Montebello facility (using SRC residues).

  7. Regulation of the AbrA1/A2 Two-Component System in Streptomyces coelicolor and the Potential of Its Deletion Strain as a Heterologous Host for Antibiotic Production

    PubMed Central

    Rico, Sergio; Yepes, Ana; Rodríguez, Héctor; Santamaría, Jorge; Antoraz, Sergio; Krause, Eva M.; Díaz, Margarita; Santamaría, Ramón I.

    2014-01-01

    The Two-Component System (TCS) AbrA1/A2 from Streptomyces coelicolor M145 is a negative regulator of antibiotic production and morphological differentiation. In this work we show that it is able to auto-regulate its expression, exerting a positive induction of its own operon promoter, and that its activation is dependent on the presence of iron. The overexpression of the abrA2 response regulator (RR) gene in the mutant ΔabrA1/A2 results in a toxic phenotype. The reason is an excess of phosphorylated AbrA2, as shown by phosphoablative and phosphomimetic AbrA2 mutants. Therefore, non-cognate histidine kinases (HKs) or small phospho-donors may be responsible for AbrA2 phosphorylation in vivo. The results suggest that in the parent strain S. coelicolor M145 the correct amount of phosphorylated AbrA2 is adjusted through the phosphorylation-dephosphorylation activity rate of the HK AbrA1. Furthermore, the ABC transporter system, which is part of the four-gene operon comprising AbrA1/A2, is necessary to de-repress antibiotic production in the TCS null mutant. Finally, in order to test the possible biotechnological applications of the ΔabrA1/A2 strain, we demonstrate that the production of the antitumoral antibiotic oviedomycin is duplicated in this strain as compared with the production obtained in the wild type, showing that this strain is a good host for heterologous antibiotic production. Thus, this genetically modified strain could be interesting for the biotechnology industry. PMID:25303210

  8. Development and Evaluation of the Habitat Demonstration Unit Medical Operations Workstation and Opportunities for Future Research

    NASA Technical Reports Server (NTRS)

    Howard, Robert L., Jr.

    2012-01-01

    As NASA develops missions to leave Earth orbit and explore distant destinations (Mars, Moon, Asteroids) it is necessary to rethink human spaceflight paradigms in the life sciences. Standards developed for low earth orbit human spaceflight may not be fully applicable and in-space research may be required to develop new standards. Preventative and emergency medical care may require new capabilities never before used in space. Due to spacecraft volume limitations, this work area may also be shared with various animal and plant life science research. This paper explores the prototype Medical Operations Workstation within the NASA Habitat Demonstration Unit and discusses some of the lessons learned from field analogue missions involving the workstation. Keywords: Exploration, medical, health, crew, injury emergency, biology, animal, plant, science, preventative, emergency.

  9. Water Use Optimization Toolset Project: Development and Demonstration Phase Draft Report

    SciTech Connect

    Gasper, John R.; Veselka, Thomas D.; Mahalik, Matthew R.; Hayse, John W.; Saha, Samrat; Wigmosta, Mark S.; Voisin, Nathalie; Rakowski, Cynthia; Coleman, Andre; Lowry, Thomas S.

    2014-05-19

    This report summarizes the results of the development and demonstration phase of the Water Use Optimization Toolset (WUOT) project. It identifies the objective and goals that guided the project, as well as demonstrating potential benefits that could be obtained by applying the WUOT in different geo-hydrologic systems across the United States. A major challenge facing conventional hydropower plants is to operate more efficiently while dealing with an increasingly uncertain water-constrained environment and complex electricity markets. The goal of this 3-year WUOT project, which is funded by the U.S. Department of Energy (DOE), is to improve water management, resulting in more energy, revenues, and grid services from available water, and to enhance environmental benefits from improved hydropower operations and planning while maintaining institutional water delivery requirements. The long-term goal is for the WUOT to be used by environmental analysts and deployed by hydropower schedulers and operators to assist in market, dispatch, and operational decisions.

  10. NASA Systems Autonomy Demonstration Project - Development of Space Station automation technology

    NASA Technical Reports Server (NTRS)

    Bull, John S.; Brown, Richard; Friedland, Peter; Wong, Carla M.; Bates, William

    1987-01-01

    A 1984 Congressional expansion of the 1958 National Aeronautics and Space Act mandated that NASA conduct programs, as part of the Space Station program, which will yield the U.S. material benefits, particularly in the areas of advanced automation and robotics systems. Demonstration programs are scheduled for automated systems such as the thermal control, expert system coordination of Station subsystems, and automation of multiple subsystems. The programs focus the R&D efforts and provide a gateway for transfer of technology to industry. The NASA Office of Aeronautics and Space Technology is responsible for directing, funding and evaluating the Systems Autonomy Demonstration Project, which will include simulated interactions between novice personnel and astronauts and several automated, expert subsystems to explore the effectiveness of the man-machine interface being developed. Features and progress on the TEXSYS prototype thermal control system expert system are outlined.

  11. Development of LM10-MIRA LOX/LNG expander cycle demonstrator engine

    NASA Astrophysics Data System (ADS)

    Rudnykh, Mikhail; Carapellese, Stefano; Liuzzi, Daniele; Arione, Luigi; Caggiano, Giuseppe; Bellomi, Paolo; D'Aversa, Emanuela; Pellegrini, Rocco; Lobov, S. D.; Gurtovoy, A. A.; Rachuk, V. S.

    2016-09-01

    This article contains results of joint works by Konstruktorskoe Buro Khimavtomatiki (KBKhA, Russia) and AVIO Company (Italy) on creation of the LM10-MIRA liquid-propellant rocket demonstrator engine for the third stage of the upgraded "Vega" launcher.Scientific and research activities conducted by KBKhA and AVIO in 2007-2014 in the frame of the LYRA Program, funded by the Italian Space Agency, with ELV as Prime contractor, and under dedicated ASI-Roscosmos inter-agencies agreement, were aimed at development and testing of a 7.5 t thrust expander cycle demonstrator engine propelled by oxygen and liquid natural gas (further referred to as LNG).

  12. Microgrid Design, Development and Demonstration - Final Report for Phase I and Phase II

    SciTech Connect

    Bose, Sumit; Krok, Michael

    2011-02-08

    This document constitutes GE’s final report for the Microgrid Design, Development and Demonstration program for DOE’s Office of Electricity Delivery and Energy Reliability, Award DE-FC02-05CH11349. It contains the final report for Phase I in Appendix I, and the results the work performed in Phase II. The program goal was to develop and demonstrate a Microgrid Energy Management (MEM) framework for a broad set of Microgrid applications that provides unified controls, protection, and energy management. This project contributed to the achievement of the U.S. Department of Energy’s Renewable and Distributed Systems Integration Program goals by developing a fully automated power delivery microgrid network that: - Reduces carbon emissions and emissions of other air pollutants through increased use of optimally dispatched renewable energy, - Increases asset use through integration of distributed systems, - Enhances reliability, security, and resiliency from microgrid applications in critical infrastructure protection, constrained areas of the electric grid, etc. - Improves system efficiency with on-site, distributed generation and improved economic efficiency through demand-side management.

  13. Development and demonstration of treatment technologies for the processing of US Department of Energy mixed waste

    SciTech Connect

    Berry, J.B.; Bloom, G.A.; Kuchynka, D.J.

    1994-06-01

    Mixed waste is defined as waste contaminated with chemically hazardous (governed by the Resource Conservation and Recovery Act) and radioactive species [governed by US Department of Energy (DOE) orders]. The Mixed Waste Integrated Program (MWIP) is responding to the need for DOE mixed waste treatment technologies that meet these dual regulatory requirements. MWIP is developing emerging and innovative treatment technologies to determine process feasibility. Technology demonstrations will be used to determine whether processes are superior to existing technologies in reducing risk, minimizing life-cycle cost, and improving process performance. The Program also provides a forum for stakeholder and customer involvement in the technology development process. MWIP is composed of six technical areas that support a mixed-waste treatment system: (1) systems analysis, (2) materials handling, (3) chemical/physical separation, (4) waste destruction and stabilization, (5) off-gas treatment, and (6) final waste form stabilization. The status of the technical initiatives and the current research, development, and demonstration in each of these areas is described in this paper.

  14. The development and testing of the Lens Antenna Deployment Demonstration (LADD) test article

    NASA Technical Reports Server (NTRS)

    Pugh, Mark L.; Denton, Robert J., Jr.; Strange, Timothy J.

    1993-01-01

    The USAF Rome Laboratory and NASA Marshall Space Flight Center, through contract to Grumman Corporation, have developed a space-qualifiable test article for the Strategic Defense Initiative Organization to demonstrate the critical structural and mechanical elements of single-axis roll-out membrane deployment for Space Based Radar (SBR) applications. The Lens Antenna Deployment Demonstration (LADD) test article, originally designed as a shuttle-attached flight experiment, is a large precision space structure which is representative of operational designs for space-fed lens antennas. Although the flight experiment was cancelled due to funding constraints and major revisions in the Strategic Defense System (SDS) architecture, development of this test article was completed in June 1989. To take full advantage of the existence of this unique structure, a series of ground tests are proposed which include static, dynamic, and thermal measurements in a simulated space environment. An equally important objective of these tests is the verification of the analytical tools used to design and develop large precision space structures.

  15. Northwest Hazardous Waste Research, Development, and Demonstration Center: Program Plan. [Contains glossary

    SciTech Connect

    Not Available

    1988-02-01

    The Northwest Hazardous Waste Research, Development, and Demonstration Center was created as part of an ongoing federal effort to provide technologies and methods that protect human health and welfare and environment from hazardous wastes. The Center was established by the Superfund Amendments and Reauthorization Act (SARA) to develop and adapt innovative technologies and methods for assessing the impacts of and remediating inactive hazardous and radioactive mixed-waste sites. The Superfund legislation authorized $10 million for Pacific Northwest Laboratory to establish and operate the Center over a 5-year period. Under this legislation, Congress authorized $10 million each to support research, development, and demonstration (RD and D) on hazardous and radioactive mixed-waste problems in Idaho, Montana, Oregon, and Washington, including the Hanford Site. In 1987, the Center initiated its RD and D activities and prepared this Program Plan that presents the framework within which the Center will carry out its mission. Section 1.0 describes the Center, its mission, objectives, organization, and relationship to other programs. Section 2.0 describes the Center's RD and D strategy and contains the RD and D objectives, priorities, and process to be used to select specific projects. Section 3.0 contains the Center's FY 1988 operating plan and describes the specific RD and D projects to be carried out and their budgets and schedules. 9 refs., 18 figs., 5 tabs.

  16. Energy Savings Potential and Research, Development, & Demonstration Opportunities for Commercial Building Appliances

    SciTech Connect

    Zogg, Robert; Goetzler, William; Ahlfeldt, Christopher; Hiraiwa, Hirokazu; Sathe, Amul; Sutherland, Timothy

    2009-12-01

    This study characterizes and assesses the appliances used in commercial buildings. The primary objectives of this study were to document the energy consumed by commercial appliances and identify research, development and demonstration (RD&D) opportunities for efficiency improvements, excluding product categories such as HVAC, building lighting, refrigeration equipment, and distributed generation systems. The study included equipment descriptions, characteristics of the equipment’s market, national energy consumption, estimates of technical potential for energy-saving technologies, and recommendations for U.S. Department of Energy programs that can promote energy savings in commercial appliances.

  17. Development of the Lens Antenna Deployment Demonstration (LADD) shuttle-attached flight experiment

    NASA Astrophysics Data System (ADS)

    Hill, H.; Johnston, D.; Frauenberger, H.

    1986-11-01

    The primary objective of the LADD Program is to develop a technology demonstration test article that can be used for both ground and flight tests to demonstrate the structural and mechanical feasibility and reliability of the single-axis roll-out space based radar (SBR) approach. As designed, the LADD will essentially be a generic strucutural experiment which incorporates all critical technology elements of the operational satellite and is applicable to a number of future antenna systems. However, to fully determine its design integrity for meeting the lens flatness and constant geometry requirements in a zero g environment under extreme thermal conditions, the LADD must be space flight tested. By accurately surveying the structure under varying conditions the membrane tolerance-holding capabilities of the structure will be demonstrated. The flight test will provide data to verify analytical tools used to predict thermal and structural behavior. Most important, the experiment will provide an initial indication of structural damping in a zero g vacuum environment. The recently completed Solar Array Flight Experiment (SAFE) showed orbital damping greater than that experienced during ground testing. From the experience and the information obtained from LADD it is hoped that designs can be confidently extrapolated to operational satellites with apertures in the 20 m by 60 m size range.

  18. Development of the Lens Antenna Deployment Demonstration (LADD) shuttle-attached flight experiment

    NASA Technical Reports Server (NTRS)

    Hill, H.; Johnston, D.; Frauenberger, H.

    1986-01-01

    The primary objective of the LADD Program is to develop a technology demonstration test article that can be used for both ground and flight tests to demonstrate the structural and mechanical feasibility and reliability of the single-axis roll-out space based radar (SBR) approach. As designed, the LADD will essentially be a generic strucutural experiment which incorporates all critical technology elements of the operational satellite and is applicable to a number of future antenna systems. However, to fully determine its design integrity for meeting the lens flatness and constant geometry requirements in a zero g environment under extreme thermal conditions, the LADD must be space flight tested. By accurately surveying the structure under varying conditions the membrane tolerance-holding capabilities of the structure will be demonstrated. The flight test will provide data to verify analytical tools used to predict thermal and structural behavior. Most important, the experiment will provide an initial indication of structural damping in a zero g vacuum environment. The recently completed Solar Array Flight Experiment (SAFE) showed orbital damping greater than that experienced during ground testing. From the experience and the information obtained from LADD it is hoped that designs can be confidently extrapolated to operational satellites with apertures in the 20 m by 60 m size range.

  19. Development of the first demonstration CFB boiler for gas and steam cogeneration

    SciTech Connect

    Fang, M; Luo, Z.; Li, X.; Wang, Q.; Shi, Z.; Ni, M.; Cen, K.

    1997-12-31

    To solve the shortage of gas and steam supply in the small towns of the country, a new gas steam cogeneration system has been developed. On the basis of the fundamental research on the system, a demonstration gas steam cogeneration system has been designed. As the phase 1 of the project, a 75t/h demonstration CFB boiler for gas steam cogeneration has been erected and operated at Yangzhong Thermal Power Plant of China. This paper introduces the first 75t/h demonstration CFB boiler for gas steam cogeneration. Due to the need of gas steam cogeneration process, the boiler has the features of high temperature cyclone separation, high solid recycle ratio, staged combustion and an external heat exchanger adjusting bed temperature and heat load. The operation results show that the boiler has wide fuel adaptability and the heating value of the coal changes from 14MJ/Kg to 25MJ/Kg. The heat load changes from 85t/h to 28t/h while steam parameter is maintained at the normal conditions. The combustion efficiency of the boiler attain 98%. The boiler design and operation experiences may be a guide to the design and operation of larger CFB units in the future.

  20. Waste management technology development and demonstration programs at Brookhaven National Laboratory

    NASA Technical Reports Server (NTRS)

    Kalb, Paul D.; Colombo, Peter

    1991-01-01

    Two thermoplastic processes for improved treatment of radioactive, hazardous, and mixed wastes were developed from bench scale through technology demonstration: polyethylene encapsulation and modified sulfur cement encapsulation. The steps required to bring technologies from the research and development stage through full scale implementation are described. Both systems result in durable waste forms that meet current Nuclear Regulatory Commission and Environmental Protection Agency regulatory criteria and provide significant improvements over conventional solidification systems such as hydraulic cement. For example, the polyethylene process can encapsulate up to 70 wt pct. nitrate salt, compared with a maximum of about 20 wt pct. for the best hydraulic cement formulation. Modified sulfur cement waste forms containing as much as 43 wt pct. incinerator fly ash were formulated, whereas the maximum quantity of this waste in hydraulic cement is 16 wt pct.

  1. Development and demonstration of treatment technologies for the processing of US Department of Energy Mixed Waste

    SciTech Connect

    Bloom, G.A.; Berry, J.B.

    1994-01-01

    Mixed waste is defined as ``waste contaminated with chemically hazardous and radioactive species.`` The Mixed Waste Integrated Program (MWIP) was established in response to the need for a unified, DOE complexwide solution to issues of mixed waste treatment that meets regulatory requirements. MWIP is developing treatment technologies that reduce risk, minimize life-cycle cost, and improve process performance as compared to existing technologies. Treatment for waste streams for which no current technology exists, and suitable waste forms for disposal, will be provided to improve operations of the DOE Office of Waste Management. MWIP is composed of six technical areas within a mixed-waste treatment system: (1) systems analysis, (2) materials handling, (3) chemical/physical separation, (4) waste destruction and stabilization, (5) off-gas treatment, and (6) final waste form stabilization. The status of the technical initiatives and the current research, development, and demonstration in each of these areas are described in this paper

  2. A data-gathering method for use in modeling energy research, development and demonstration programs

    NASA Astrophysics Data System (ADS)

    Meyer, M. A.; Booker, J. M.; Cullingford, H. S.; Peaslee, A. T., Jr.

    The development and testing of a data-gathering method for use in a computer program designed to model energy research, development, and demonstration programs for decisionmakers are described. The data-gathering method consists of face-to-face interviews with the scientists working on the projects that will be modeled by the computer program. The basic information gained from an interview includes time estimates for reaching certain project goals and the probability of achieving those goals within the times estimated. The interview method is based on decision analysis techniques. The Magnetic Fusion Energy program of the US Department of Energy was selected as the test case. The data gathering method was used at five fusion projects to determine whether it could meet its design criteria. Extensive statistical analysis was performed to learn how much the expert's answers agreed, what factors were likely to enter into their estimates, and how their estimates corresponded.

  3. Waste management technology development and demonstration programs at Brookhaven National Laboratory

    SciTech Connect

    Kalb, P.D.; Colombo, P.

    1991-12-31

    Two thermoplastic processes for improved treatment of radioactive, hazardous, and mixed wastes have been developed from bench-scale through technology demonstration: polyethylene encapsulation and modified sulfur cement encapsulation. The steps required to bring technologies from the research and development stage through full-scale implementation are described. Both systems result in durable waste forms that meet current Nuclear Regulatory Commission and Environmental Protection Agency regulatory criteria and provide significant improvements over conventional solidification systems such as hydraulic cement. For example, the polyethylene process can encapsulate up to 70 wt % nitrate salt, compared with a maximum of about 20 wt % for the best hydraulic cement formulation. Modified sulfur cement waste forms containing as much as 43 wt % incinerator fly ash have been formulated, whereas the maximum quantity of this waste in hydraulic cement is 16 wt %.

  4. A Summary fo Solar Sail Technology Developments and Proposed Demonstration Missions

    NASA Technical Reports Server (NTRS)

    Garner, Charles; Diedrich, Benjamin; Leipold, Manfred

    1999-01-01

    NASA's drive to reduce mission costs and accept the risk of incorporating innovative, high payoff technologies into it's missions while simultaneously undertaking ever more difficult missions has sparked a greatly renewed interest in solar sails. From virtually no technology or flight mission studies activity three years ago solar sails are now included in NOAA, NASA, DOD, DLR, ESA and ESTEC technology development programs and technology roadmaps. NASA programs include activities at Langley Research Center, Jet Propulsion Laboratory, Marshall Space Flight Center, Goddard Space Flight Center, and the NASA Institute for Advanced Concepts; NOAA has received funding for a proposed solar sail mission; DLR is designing and fabricating a 20-m laboratory model sail, there are four demonstration missions under study at industry, NASA, DOD and Europe, two new text books on solar sailing were recently published and one new test book is planned. This paper summarizes these on-going developments in solar sails.

  5. Pit disassembly and conversion demonstration environmental assessment and research and development activities

    SciTech Connect

    1998-08-01

    A significant portion of the surplus plutonium is in the form of pits, a nuclear weapons component. Pits are composed of plutonium which is sealed in a metallic shell. These pits would need to be safely disassembled and permanently converted to an unclassified form that would be suitable for long-term disposition and international inspection. To determine the feasibility of an integrated pit disassembly and conversion system, a Pit Disassembly and Conversion Demonstration is proposed to take place at the Los Alamos National Laboratory (LANL). This demonstration would be done in existing buildings and facilities, and would involve the disassembly of up to 250 pits and conversion of the recovered plutonium to plutonium metal ingots and plutonium dioxide. This demonstration also includes the conversion of up to 80 kilograms of clean plutonium metal to plutonium dioxide because, as part of the disposition process, some surplus plutonium metal may be converted to plutonium dioxide in the same facility as the surplus pits. The equipment to be used for the proposed demonstration addressed in this EA would use some parts of the Advanced Recovery and Integrated Extraction System (ARIES) capability, other existing equipment/capacities, plus new equipment that was developed at other sites. In addition, small-scale R and D activities are currently underway as part of the overall surplus plutonium disposition program. These R and D activities are related to pit disassembly and conversion, MOX fuel fabrication, and immobilization (in glass and ceramic forms). They are described in Section 7.0. On May 16, 1997, the Office of Fissile Materials Disposition (MD) notified potentially affected states and tribes that this EA would be prepared in accordance with NEPA. This EA has been prepared to provide sufficient information for DOE to determine whether a Finding of No Significant Impact (FONSI) is warranted or whether an EIS must be prepared.

  6. Initial Development and Pilot Study Design of Interactive Lecture Demonstrations for ASTRO 101

    NASA Astrophysics Data System (ADS)

    Schwortz, Andria C.; French, D. A; Gutierrez, Joseph V; Sanchez, Richard L; Slater, Timothy F.; Tatge, Coty

    2014-06-01

    Interactive lecture demonstrations (ILDs) have repeatedly shown to be effective tools for improving student achievement in the context of learning physics. As a first step toward systematic development of interactive lecture demonstrations in ASTRO 101, the introductory astronomy survey course, a systematic review of education research, describing educational computer simulations (ECSs) reveals that initial development requires a targeted study of how ASTRO 101 students respond to ECSs in the non-science majoring undergraduate lecture setting. In this project we have adopted the process by which ILDs were designed, pilot-tested, and successfully implemented in the context of physics teaching (Sokoloff & Thornton, 1997; Sokoloff & Thornton, 2004). We have designed the initial pilot-test set of ASTRO 101 ILD instructional materials relying heavily on ECSs. Both an instructor’s manual and a preliminary classroom-ready student workbook have been developed, and we are implementing a pilot study to explore their effectiveness in communicating scientific content, and the extent to which they might enhance students’ knowledge of and perception about astronomy and science in general. The study design uses a pre-/post-test quasi-experimental study design measuring students’ normalized gain scores, calculated as per Hake (1998) and Prather (2009), using a slightly modified version of S. Slater’s (2011) Test Of Astronomy STandards TOAST combined with other instruments. The results of this initial study will guide the iterative development of ASTRO 101 ILDs that are intended to both be effective at enhancing student achievement and easy for instructors to successfully implement.

  7. Nevada Test Site-Directed Research, Development, and Demonstration. FY2005 report

    SciTech Connect

    Lewis, Will

    2006-09-01

    The Nevada Test Site-Directed Research, Development, and Demonstration (SDRD) program completed a very successful year of research and development activities in FY 2005. Fifty new projects were selected for funding this year, and five FY 2004 projects were brought to conclusion. The total funds expended by the SDRD program were $5.4 million, for an average per project cost of just under $100,000. Two external audits of SDRD accounting practices were conducted in FY 2005. Both audits found the program's accounting practices consistent with the requirements of DOE Order 413.2A, and one included the observation that the NTS contractor ''did an exceptional job in planning and executing year-start activities.'' Highlights for the year included: the filing of 18 invention disclosures for intellectual property generated by FY 2005 projects; programmatic adoption of 17 FY 2004 SDRD-developed technologies; participation in the tri-lab Laboratory Directed Research and Development (LDRD) and SDRD program review that was broadly attended by NTS, NNSA, LDRD, and U.S. Department of Homeland Security representatives; peer reviews of all FY 2005 projects; and the successful completion of 55 R&D projects, as presented in this report.

  8. Two-Phase Flow Technology Developed and Demonstrated for the Vision for Exploration

    NASA Technical Reports Server (NTRS)

    Sankovic, John M.; McQuillen, John B.; Lekan, Jack F.

    2005-01-01

    NASA s vision for exploration will once again expand the bounds of human presence in the universe with planned missions to the Moon and Mars. To attain the numerous goals of this vision, NASA will need to develop technologies in several areas, including advanced power-generation and thermal-control systems for spacecraft and life support. The development of these systems will have to be demonstrated prior to implementation to ensure safe and reliable operation in reduced-gravity environments. The Two-Phase Flow Facility (T(PHI) FFy) Project will provide the path to these enabling technologies for critical multiphase fluid products. The safety and reliability of future systems will be enhanced by addressing focused microgravity fluid physics issues associated with flow boiling, condensation, phase separation, and system stability, all of which are essential to exploration technology. The project--a multiyear effort initiated in 2004--will include concept development, normal-gravity testing (laboratories), reduced gravity aircraft flight campaigns (NASA s KC-135 and C-9 aircraft), space-flight experimentation (International Space Station), and model development. This project will be implemented by a team from the NASA Glenn Research Center, QSS Group, Inc., ZIN Technologies, Inc., and the Extramural Strategic Research Team composed of experts from academia.

  9. Research, Development and Demonstration of Micro-CHP System for Residential Applications

    SciTech Connect

    Karl Mayer

    2010-03-31

    ECR International and its joint venture company, Climate Energy, are at the forefront of the effort to deliver residential-scale combined heat and power (Micro-CHP) products to the USA market. Part of this substantial program is focused on the development of a new class of steam expanders that offers the potential for significantly lower costs for small-scale power generation technology. The heart of this technology is the scroll expander, a machine that has revolutionized the HVAC refrigerant compressor industry in the last 15 years. The liquid injected cogeneration (LIC) technology is at the core of the efforts described in this report, and remains an excellent option for low cost Micro-CHP systems. ECR has demonstrated in several prototype appliances that the concept for LIC can be made into a practical product. The continuing challenge is to identify economical scroll machine designs that will meet the performance and endurance requirements needed for a long life appliance application. This report describes the numerous advances made in this endeavor by ECR International. Several important advances are described in this report. Section 4 describes a marketing and economics study that integrates the technical performance of the LIC system with real-world climatic data and economic analysis to assess the practical impact that different factors have on the economic application of Micro-CHP in residential applications. Advances in the development of a working scroll steam expander are discussed in Section 5. A rigorous analytical assessment of the performance of scroll expanders, including the difficult to characterize impact of pocket to pocket flank leakage, is presented in Section 5.1. This is followed with an FEA study of the thermal and pressure induced deflections that would result from the normal operation of an advanced scroll expander. Section 6 describes the different scroll expanders and test fixtures developed during this effort. Another key technical

  10. Development of a UAV-based Global Ozone Lidar Demonstrator (GOLD)

    NASA Astrophysics Data System (ADS)

    Browell, E. V.; Deyoung, R. J.; Hair, J. W.; Ismail, S.; McGee, T.; Hardesty, R. M.; Brewer, W. A.; McDermid, I. S.

    2006-12-01

    Global ozone measurements are needed across the troposphere with high vertical resolution to enable comprehensive studies of continental and intercontinental atmospheric chemistry and dynamics, which are affected by diverse natural and human-induced processes. The development of a unattended aerial vehicle (UAV) based Global Ozone Lidar Demonstrator (GOLD) is an important step in enabling a space-based ozone and aerosol lidar and for conducting unique UAV-based large-scale atmospheric investigations. The GOLD system will incorporate the most advanced technology developed under the NASA Laser Risk Reduction Program (LRRP) and the Small Business Innovative Research (SBIR) program to produce a compact, autonomously operating ozone and aerosol Differential Absorption Lidar (DIAL) system for a UAV platform. This system will leverage advanced Nd:YAG and optical parametric oscillator (OPO) laser technologies being developed by ITT Industries under the LRRP and the autonomously operating ozone DIAL system being developed by Science and Engineering Services Inc. (SESI) under an SBIR Phase-3 contract. Laser components from ITT will be integrated into the SESI DIAL system, and the resulting GOLD system will be flight tested on a NASA UAV. The development of the GOLD system was initiated as part of the NASA Instrument Incubator Program in December 2005, and great progress has been made towards completing major GOLD subsystems. ITT has begun construction of the high-power Nd:YAG pump laser and the ultraviolet OPO for generating the ozone DIAL wavelengths of 290 and 300 nm and the aerosol visible wavelength at 532 nm. SESI is completing the Phase-3 SBIR contract for the delivery and demonstration of the ozone DIAL receiver and data system, and NOAA is completing detector evaluations for use in the GOLD system. Welch Mechanical is examining system designs for integrating GOLD into the external pod that will be hung under the new IKANA (Predator-B) UAV that NASA Dryden is

  11. MicroRNA in late lung development and bronchopulmonary dysplasia: the need to demonstrate causality.

    PubMed

    Nardiello, Claudio; Morty, Rory E

    2016-12-01

    MicroRNA are emerging as powerful regulators of cell differentiation and tissue and organ development. Several microRNA have been described to play a role in branching morphogenesis, a key step in early lung development. However, considerably less attention has been paid to microRNA as regulators of the process of secondary septation, which drives lung alveolarization during late lung development. Secondary septation is severely perturbed in bronchopulmonary dysplasia (BPD), a common complication of preterm birth characterized by blunted alveolarization. A number of studies to date have reported microRNA microarray screens in animal models of BPD; however, only two studies have attempted to demonstrate causality. Although the expression of miR-150 was altered in experimental BPD, a miR-150(-/-) knockout mouse did not exhibit appreciable protection in a BPD animal model. Similarly, while the expression of miR-489 in the lung was reduced in clinical and experimental BPD, antagomiR and over-expression approaches could not validate a role for miR-489 in the impaired alveolarization associated with experimental BPD. This mini-review aims to highlight microRNA that have been revealed by multiple microarray studies to be potential causal players in normal and pathological alveolarization. Additionally, the challenges faced in attempting to demonstrate a causal role for microRNA in lung alveolarization are discussed. These include the tremendous variability in the animal models employed, and the limitations and advantages offered by the available tools, including antagomiRs and approaches for the validation of a specific microRNA-mRNA interaction during lung alveolarization. PMID:27216745

  12. Industrial advanced turbine systems: Development and demonstration. Quarterly report, January 1--March 31, 1998

    SciTech Connect

    1998-08-01

    The US Department of Energy (DOE) has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The objective of the cooperative agreements granted under the program is to join the DOE with industry in research and development that will lead to commercial offerings in the private sector. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The ATS will foster (1) early market penetration that enhances the global competitiveness of US industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace, and (4) the retention and expansion of the skilled US technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program. Phase 3 of the work is separated into two subphases: Phase 3A entails Component Design and Development; Phase 3B will involve Integrated Subsystem Testing. Phase 4 will cover Host Site Testing. As of the end of the reporting period work on the program is 29.1% complete (24.7% last quarter). Work on the Mercury 50 development and ATS technology development portions of the program (WBS 10000 et seq) is 48.9% complete (41.6% last quarter). Estimates of percent complete are based upon milestones completed. In order to maintain objectivity in assessing schedule progress, Solar uses a 0/100 percent complete assumption for milestones rather than subjectively estimating progress toward completion of milestones. Cost and schedule variance information is provided in Section 4.0 Program Management.

  13. Promoter determining the timing and spatial localization of transcription of a cloned Streptomyces coelicolor gene encoding a spore-associated polypeptide.

    PubMed Central

    Guijarro, J; Santamaria, R; Schauer, A; Losick, R

    1988-01-01

    Streptomyces coelicolor is a filamentous, gram-positive bacterium that exhibits a complex cycle of morphological differentiation involving the formation of an aerial mycelium of multinucleoid hyphae which undergo septation to form long chains of spores. We report the identification of two proteins of 13 and 3 kilodaltons, designated SapA and SapB, respectively, that are produced during formation of the aerial mycelium and are found in assocation with purified, mature spores. We cloned the structural gene (sapA) for one of these spore-associated proteins. Nucleotide sequence analysis suggests that the 13-kilodalton polypeptide is derived from a larger pre- or preproprotein containing a leader sequence of 37 amino acids. Nuclease protection-hybridization analysis and experiments using the Vibrio harveyi, luciferase-encoding luxAB operon as a gene tag demonstrated that expression of sapA is controlled from a promoter contained within a region of less than 110 base pairs in length, whose transcription start site is located approximately 50 base pairs upstream from the initiation codon for the sapA open reading frame. Transcription of sapA was induced at the time of appearance of the aerial mycelium, and the level of sapA transcripts was significantly reduced in certain mutants blocked in aerial mycelium (bld) and or spore (whi) formation. As further evidence of the association of sapA transcription with morphological differentiation, experiments in which we monitored sapA transcription topographically by use of a sapA-luxAB operon fusion demonstrated a close spatial correlation between colony regions undergoing aerial mycelium formation and zones of sapA-promoted light emission. Images PMID:2450872

  14. Comparative genomics of transport proteins in developmental bacteria: Myxococcus xanthus and Streptomyces coelicolor

    PubMed Central

    2013-01-01

    Background Two of the largest fully sequenced prokaryotic genomes are those of the actinobacterium, Streptomyces coelicolor (Sco), and the δ-proteobacterium, Myxococcus xanthus (Mxa), both differentiating, sporulating, antibiotic producing, soil microbes. Although the genomes of Sco and Mxa are the same size (~9 Mbp), Sco has 10% more genes that are on average 10% smaller than those in Mxa. Results Surprisingly, Sco has 93% more identifiable transport proteins than Mxa. This is because Sco has amplified several specific types of its transport protein genes, while Mxa has done so to a much lesser extent. Amplification is substrate- and family-specific. For example, Sco but not Mxa has amplified its voltage-gated ion channels but not its aquaporins and mechano-sensitive channels. Sco but not Mxa has also amplified drug efflux pumps of the DHA2 Family of the Major Facilitator Superfamily (MFS) (49 versus 6), amino acid transporters of the APC Family (17 versus 2), ABC-type sugar transport proteins (85 versus 6), and organic anion transporters of several families. Sco has not amplified most other types of transporters. Mxa has selectively amplified one family of macrolid exporters relative to Sco (16 versus 1), consistent with the observation that Mxa makes more macrolids than does Sco. Conclusions Except for electron transport carriers, there is a poor correlation between the types of transporters found in these two organisms, suggesting that their solutions to differentiative and metabolic needs evolved independently. A number of unexpected and surprising observations are presented, and predictions are made regarding the physiological functions of recognizable transporters as well as the existence of yet to be discovered transport systems in these two important model organisms and their relatives. The results provide insight into the evolutionary processes by which two dissimilar prokaryotes evolved complexity, particularly through selective chromosomal gene

  15. Extracellular complementation and the identification of additional genes involved in aerial mycelium formation in Streptomyces coelicolor.

    PubMed

    Nodwell, J R; Yang, M; Kuo, D; Losick, R

    1999-02-01

    Morphogenesis in the bacterium Streptomyces coelicolor involves the formation of a lawn of hair-like aerial hyphae on the colony surface that stands up in the air and differentiates into chains of spores. bld mutants are defective in the formation of this aerial mycelium and grow as smooth, hairless colonies. When certain pairs of bld mutants are grown close to one another on rich sporulation medium, they exhibit extracellular complementation such that one mutant restores aerial mycelium formation to the other. The extracellular complementation relationships of most of the previously isolated bld mutants placed them in a hierarchy of extracellular complementation groups. We have screened for further bld mutants with precautions intended to maximize the discovery of additional genes. Most of the 50 newly isolated mutant strains occupy one of three of the previously described positions in the hierarchy, behaving like bldK, bldC, or bldD mutants. We show that the mutations in some of the strains that behave like bldK are bldK alleles but that others fall in a cluster at a position on the chromosome distinct from that of any known bld gene. We name this locus bldL. By introducing cloned genes into the strains that exhibit bldC or bldD-like extracellular complementation phenotypes, we show that most of these strains are likely to contain mutations in genes other than bldC or bldD. These results indicate that the genetic control of aerial mycelium formation is more complex than previously recognized and support the idea that a high proportion of bld genes are directly or indirectly involved in the production of substances that are exchanged between cells during morphological differentiation. PMID:9927452

  16. Structural analysis of the catalytic mechanism and stereoselectivity in Streptomyces coelicolor alditol oxidase.

    PubMed

    Forneris, Federico; Heuts, Dominic P H M; Delvecchio, Manuela; Rovida, Stefano; Fraaije, Marco W; Mattevi, Andrea

    2008-01-22

    Alditol oxidase (AldO) from Streptomyces coelicolor A3(2) is a soluble monomeric flavin-dependent oxidase that performs selective oxidation of the terminal primary hydroxyl group of several alditols. Here, we report the crystal structure of the recombinant enzyme in its native state and in complex with both six-carbon (mannitol and sorbitol) and five-carbon substrates (xylitol). AldO shares the same folding topology of the members of the vanillyl-alcohol oxidase family of flavoenzymes and exhibits a covalently linked FAD which is located at the bottom of a funnel-shaped pocket that forms the active site. The high resolution of the three-dimensional structures highlights a well-defined hydrogen-bonding network that tightly constrains the substrate in the productive conformation for catalysis. Substrate binding occurs through a lock-and-key mechanism and does not induce conformational changes with respect to the ligand-free protein. A network of charged residues is proposed to favor catalysis through stabilization of the deprotonated form of the substrate. A His side chain acts as back door that "pushes" the substrate-reactive carbon atom toward the N5-C4a locus of the flavin. Analysis of the three-dimensional structure reveals possible pathways for diffusion of molecular oxygen and a small cavity on the re side of the flavin that may host oxygen during FAD reoxidation. These features combined with the tight shape of the catalytic site provide insights into the mechanism of AldO-mediated regioselective oxidation reactions and its substrate specificity. PMID:18154360

  17. Extracellular complementation and the identification of additional genes involved in aerial mycelium formation in Streptomyces coelicolor.

    PubMed Central

    Nodwell, J R; Yang, M; Kuo, D; Losick, R

    1999-01-01

    Morphogenesis in the bacterium Streptomyces coelicolor involves the formation of a lawn of hair-like aerial hyphae on the colony surface that stands up in the air and differentiates into chains of spores. bld mutants are defective in the formation of this aerial mycelium and grow as smooth, hairless colonies. When certain pairs of bld mutants are grown close to one another on rich sporulation medium, they exhibit extracellular complementation such that one mutant restores aerial mycelium formation to the other. The extracellular complementation relationships of most of the previously isolated bld mutants placed them in a hierarchy of extracellular complementation groups. We have screened for further bld mutants with precautions intended to maximize the discovery of additional genes. Most of the 50 newly isolated mutant strains occupy one of three of the previously described positions in the hierarchy, behaving like bldK, bldC, or bldD mutants. We show that the mutations in some of the strains that behave like bldK are bldK alleles but that others fall in a cluster at a position on the chromosome distinct from that of any known bld gene. We name this locus bldL. By introducing cloned genes into the strains that exhibit bldC or bldD-like extracellular complementation phenotypes, we show that most of these strains are likely to contain mutations in genes other than bldC or bldD. These results indicate that the genetic control of aerial mycelium formation is more complex than previously recognized and support the idea that a high proportion of bld genes are directly or indirectly involved in the production of substances that are exchanged between cells during morphological differentiation. PMID:9927452

  18. Development/Demonstration of an Advanced Oxy-Fuel Front-End System

    SciTech Connect

    Mighton, Steven, J.

    2007-08-06

    Owens Corning and other glass manufacturers have used oxy-fuel combustion technology successfully in furnaces to reduce emissions, increase throughput, reduce fuel consumption and, depending on the costs of oxygen and fuel, reduce energy costs. The front end of a fiberglass furnace is the refractory channel system that delivers glass from the melter to the forming process. After the melter, it is the second largest user of energy in a fiberglass plant. A consortium of glass companies and suppliers, led by Owens Corning, was formed to develop and demonstrate oxy/fuel combustion technology for the front end of a fiberglass melter, to demonstrate the viability of this energy saving technology to the U.S. glass industry, as a D.O.E. sponsored project. The project goals were to reduce natural gas consumption and CO2 green house gas emissions by 65 to 70% and create net cost savings after the purchase of oxygen to achieve a project payback of less than 2 years. Project results in Jackson, TN included achieving a 56% reduction in gas consumption and CO2 emissions. A subsequent installation in Guelph ON, not impacted by unrelated operational changes in Jackson, achieved a 64% reduction. Using the more accurate 64% reduction in the payback calculation yielded a 2.2 year payback in Jackson. The installation of the demonstration combustion system saves 77,000 DT/yr of natural gas or 77 trillion Btu/yr and eliminates 4500 tons/yr of CO2 emissions. This combustion system is one of several energy and green house gas reduction technologies being adopted by Owens Corning to achieve aggressive goals relating to the company’s global facility environmental footprint.

  19. Industrial advanced turbine systems: Development and demonstration. Quarterly report, October 1--December 31, 1997

    SciTech Connect

    1998-06-01

    The US DOE has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program. Phase 3 of the work is separated into two subphases: Phase 3A entails Component Design and Development; Phase 3B will involve Integrated Subsystem Testing. Phase 4 will cover Host Site Testing. Forecasts call for completion of the program within budget as originally estimated. Scheduled completion is forecasted to be approximately 3 years late to original plan. This delay has been intentionally planned in order to better match program tasks to the anticipated availability of DOE funds. To ensure the timely realization of DOE/Solar program goals, the development schedule for the smaller system (Mercury 50) and enabling technologies has been maintained, and commissioning of the field test unit is scheduled for May of 2000. As of the end of the reporting period work on the program is 24.7% complete (22.8% last quarter). Work on the Mercury 50 development and ATS technology development portions of the program (WBS 10000 et seq) is 41.6% complete. Although a great amount of work occurred in the quarter, a significant amount of this work entailed the revision and rerelease of several Mercury 50 drawings. Estimates of percent compete are based upon milestones completed. In order to maintain objectivity in assessing schedule progress, Solar uses a 0/100 percent complete assumption for milestones rather than subjectively estimating progress toward completion of milestones. Cost and schedule variation information is provided in Section 4.0 Program Management.

  20. Industrial Advanced Turbine Systems: Development and Demonstration. Annual report, September 14, 1995--September 30, 1996

    SciTech Connect

    1998-12-31

    The U.S. Department of Energy (DOE) has initiated a program for advanced turbine systems (ATS) that will serve industrial power generation markets. The objective of the cooperative agreements granted under the program is to join the DOE with industry in research and development that will lead to commercial offerings in the private sector. The ATS will provide ultra-high efficiency, environmental superiority, and cost competitiveness. The ATS will foster (1) early market penetration that enhances the global competitiveness of U.S. industry, (2) public health benefits resulting from reduced exhaust gas emissions of target pollutants, (3) reduced cost of power used in the energy-intensive industrial marketplace and (4) the retention and expansion of the skilled U.S. technology base required for the design, development and maintenance of state-of-the-art advanced turbine products. The Industrial ATS Development and Demonstration program is a multi-phased effort. Solar Turbines Incorporated (Solar) has participated in Phases 1 and 2 of the program. On September 14, 1995 Solar was awarded a Cooperative Agreement for Phases 3 and 4 of the program (DE-FC21-95MC31173) by the DOE`s Office of Energy Efficiency and Renewable Energy (EE). Technical administration of the Cooperative Agreement will be provided from EE`s Chicago Operations Office. Contract administration of the Cooperative Agreement will be provided from DOE`s Office of Fossil Energy, Morgantown Energy Technology Center (METC).

  1. Development and pilot demonstration program of a waste minimization plan at Argonne National Laboratory

    SciTech Connect

    Peters, R.W.; Wentz, C.A.; Thuot, J.R.

    1991-01-01

    In response to US Department of Energy directives, Argonne National Laboratory (ANL) has developed a waste minimization plan aimed at reducing the amount of wastes at this national research and development laboratory. Activities at ANL are primarily research- oriented and as such affect the amount and type of source reduction that can be achieved at this facility. The objective of ANL's waste minimization program is to cost-effectively reduce all types of wastes, including hazardous, mixed, radioactive, and nonhazardous wastes. The ANL Waste Minimization Plan uses a waste minimization audit as a systematic procedure to determine opportunities to reduce or eliminate waste. To facilitate these audits, a computerized bar-coding procedure is being implemented at ANL to track hazardous wastes from where they are generated to their ultimate disposal. This paper describes the development of the ANL Waste Minimization Plan and a pilot demonstration of the how the ANL Plan audited the hazardous waste generated within a selected divisions of ANL. It includes quantitative data on the generation and disposal of hazardous waste at ANL and describes potential ways to minimize hazardous wastes. 2 refs., 5 figs., 8 tabs.

  2. Development and demonstration of a wood-fired gas turbine system

    SciTech Connect

    Sethi, V.

    1997-10-01

    Power Generating Inc. (PGI) has developed and patented a unique direct-fired gas turbine power system (PGI Power System) that operates on solid wood-based fuels. The PGI Power System is designed to generate from 500 kilowatts to 3.5 megawatts of electrical power and up to 30 million Btu per hour of thermal energy for various industrial and utility applications. The system is expected to operate at thermal efficiency levels greater than 70% through full utilization of both the electrical and thermal energy it generates at a specific host facility. PGI and WRI built a 450-kW prototype system at the Western Research Institute (WRI) facilities in Laramie, Wyoming, to demonstrate the technical and economic viability of the PGI Power System. The plant has undergone a brief shakedown, and is presently being operated on white wood. In previous attempts to develop similar systems, the major technical hindrance to long-term operation of a gas turbine power system has been degradation of the hot section in the gas turbine. This problem is overcome in the PGI Power System through its unique design, by closely controlling fuel specifications, and by developing specialized operating procedures. In wood-fired testing conducted to date, no degradation in the engine performance is obvious.

  3. Experimental Development and Demonstration of Ultrasonic Measurement Diagnostics for Sodium Fast Reactor Thermal-hydraulics

    SciTech Connect

    Tokuhiro, Akira; Jones, Byron

    2013-09-13

    This research project will address some of the principal technology issues related to sodium-cooled fast reactors (SFR), primarily the development and demonstration of ultrasonic measurement diagnostics linked to effective thermal convective sensing under normatl and off-normal conditions. Sodium is well-suited as a heat transfer medium for the SFR. However, because it is chemically reactive and optically opaque, it presents engineering accessibility constraints relative to operations and maintenance (O&M) and in-service inspection (ISI) technologies that are currently used for light water reactors. Thus, there are limited sensing options for conducting thermohydraulic measurements under normal conditions and off-normal events (maintenance, unanticipated events). Acoustic methods, primarily ultrasonics, are a key measurement technology with applications in non-destructive testing, component imaging, thermometry, and velocimetry. THis project would have yielded a better quantitative and qualitative understanding of the thermohydraulic condition of solium under varied flow conditions. THe scope of work will evaluate and demonstrate ultrasonic technologies and define instrumentation options for the SFR.

  4. Research, development, and demonstration of nickel-iron batteries for electric vehicle propulsion. Annual report, 1980

    SciTech Connect

    Not Available

    1981-03-01

    The objective of the Eagle-Picher nickel-iron battery program is to develop a nickel-iron battery for use in the propulsion of electric and electric-hybrid vehicles. To date, the program has concentrated on the characterization, fabrication and testing of the required electrodes, the fabrication and testing of full-scale cells, and finally, the fabrication and testing of full-scale (270 AH) six (6) volt modules. Electrodes of the final configuration have now exceeded 1880 cycles and are showing minimal capacity decline. Full-scale cells have presently exceeded 600 cycles and are tracking the individual electrode tests almost identically. Six volt module tests have exceeded 500 cycles, with a specific energy of 48 Wh/kg. Results to date indicate the nickel-iron battery is beginning to demonstrate the performance required for electric vehicle propulsion.

  5. Hybrid Neural-Network: Genetic Algorithm Technique for Aircraft Engine Performance Diagnostics Developed and Demonstrated

    NASA Technical Reports Server (NTRS)

    Kobayashi, Takahisa; Simon, Donald L.

    2002-01-01

    As part of the NASA Aviation Safety Program, a unique model-based diagnostics method that employs neural networks and genetic algorithms for aircraft engine performance diagnostics has been developed and demonstrated at the NASA Glenn Research Center against a nonlinear gas turbine engine model. Neural networks are applied to estimate the internal health condition of the engine, and genetic algorithms are used for sensor fault detection, isolation, and quantification. This hybrid architecture combines the excellent nonlinear estimation capabilities of neural networks with the capability to rank the likelihood of various faults given a specific sensor suite signature. The method requires a significantly smaller data training set than a neural network approach alone does, and it performs the combined engine health monitoring objectives of performance diagnostics and sensor fault detection and isolation in the presence of nominal and degraded engine health conditions.

  6. 3-d finite element model development for biomechanics: a software demonstration

    SciTech Connect

    Hollerbach, K.; Hollister, A.M.; Ashby, E.

    1997-03-01

    Finite element analysis is becoming an increasingly important part of biomechanics and orthopedic research, as computational resources become more powerful, and data handling algorithms become more sophisticated. Until recently, tools with sufficient power did not exist or were not accessible to adequately model complicated, three-dimensional, nonlinear biomechanical systems. In the past, finite element analyses in biomechanics have often been limited to two-dimensional approaches, linear analyses, or simulations of single tissue types. Today, we have the resources to model fully three-dimensional, nonlinear, multi-tissue, and even multi-joint systems. The authors will present the process of developing these kinds of finite element models, using human hand and knee examples, and will demonstrate their software tools.

  7. Identification of sites for the low-level waste disposal development and demonstration program

    SciTech Connect

    Ketelle, R.H.; Lee, D.W.

    1988-04-01

    This report presents the results of site selection studies for potential low-level radioactive waste disposal sites on the Oak Ridge Reservation (ORR). Summaries of the site selection procedures used and results of previous site selection studies on the ORR are included. This report includes recommendations of sites for demonstration of shallow land burial using engineered trench designs and demonstration of above-grade disposal using design concepts similar to those used in tumulus disposal. The site selection study, like its predecessor (ORNL/TM-9717, Use of DOE Site Selection Criteria for Screening Low-Level Waste Disposal Sites on the Oak Ridge Reservation), involved application of exclusionary site screening criteria to the region of interest to eliminate unacceptable areas from consideration. Also like the previous study, the region of interest for this study was limited to the Oak Ridge Department of Energy Reservation. Reconnaissance-level environmental data were used in the study, and field inspections of candidate sites were made to verify the available reconnaissance data. Five candidate sites, all underlain by Knox dolomite residuum and bedrock, were identified for possible development of shallow land burial facilities. Of the five candidate sites, the West Chestnut site was judged to be best suited for deployment of the shallow land burial technology. Three candidate sites, all underlain by the Conasauga Group in Bear Creek Valley, were identified for possible development of above-grade disposal technologies. Of the three sites identified, the Central Bear Creek Valley site lying between State Route 95 and Gum Hollow Road was ranked most favorable for deployment of the above-grade disposal technology.

  8. A Regulatory Gene SCO2140 is Involved in Antibiotic Production and Morphological Differentiation of Streptomyces coelicolor A3(2).

    PubMed

    Yu, Lingjun; Pan, Yuanyuan; Liu, Gang

    2016-08-01

    Streptomyces coelicolor is the soil-dwelling bacterium with a complex life cycle and a strong ability to produce plenty of secondary metabolites which are strictly regulated by a variety of regulators. Amino acid alignment shows that the deduced protein of SCO2140 belongs to the family of Leucine-responsive regulatory proteins (Lrps). Disruption of SCO2140 significantly decreased the yields of actinorhodin and calcium-dependent antibiotics, and the complemented strain restored the antibiotic productions to the wild-type level. In contrast, overexpression of SCO2140 increased the actinorhodin production. In agreement with it, the transcriptions of actII-ORF4 and cdaR remarkably reduced in the SCO2140 disruption mutant. The aerial mycelium formation of the SCO2140 disruption mutant was clearly delayed in R2YE medium due to the decrease of ramS expression while its complemented strain could restore the normal formation of aerial mycelia. These results indicated that SCO2140 was involved in antibiotic biosynthesis and morphological differentiation of Streptomyces coelicolor A3(2). PMID:27113590

  9. ScbR- and ScbR2-mediated signal transduction networks coordinate complex physiological responses in Streptomyces coelicolor

    PubMed Central

    Li, Xiao; Wang, Juan; Li, Shanshan; Ji, Junjie; Wang, Weishan; Yang, Keqian

    2015-01-01

    In model organism Streptomyces coelicolor, γ-butyrolactones (GBLs) and antibiotics were recognized as signalling molecules playing fundamental roles in intra- and interspecies communications. To dissect the GBL and antibiotic signalling networks systematically, the in vivo targets of their respective receptors ScbR and ScbR2 were identified on a genome scale by ChIP-seq. These identified targets encompass many that are known to play important roles in diverse cellular processes (e.g. gap1, pyk2, afsK, nagE2, cdaR, cprA, cprB, absA1, actII-orf4, redZ, atrA, rpsL and sigR), and they formed regulatory cascades, sub-networks and feedforward loops to elaborately control key metabolite processes, including primary and secondary metabolism, morphological differentiation and stress response. Moreover, interplay among ScbR, ScbR2 and other regulators revealed intricate cross talks between signalling pathways triggered by GBLs, antibiotics, nutrient availability and stress. Our work provides a global view on the specific responses that could be triggered by GBL and antibiotic signals in S. coelicolor, among which the main echo was the change of production profile of endogenous antibiotics and antibiotic signals manifested a role to enhance bacterial stress tolerance as well, shedding new light on GBL and antibiotic signalling networks widespread among streptomycetes. PMID:26442964

  10. Crystallization and preliminary X-ray analysis of AbsC, a novel regulator of antibiotic production in Streptomyces coelicolor

    SciTech Connect

    Stevenson, Clare E. M.; Kock, Holger; Mootien, Saraspadee; Davies, Sîan C.; Bibb, Mervyn J.; Lawson, David M.

    2007-03-01

    A novel regulator of antibiotic production in S. coelicolor, AbsC, has been crystallized in space group P2{sub 1}2{sub 1}2{sub 1}. X-ray data to 2.25 Å resolution were collected on station PX 14.1 at Daresbury. Crystals of recombinant AbsC (subunit MW = 18 313 Da; 158 amino acids), a novel regulator of antibiotic production from Streptomyces coelicolor, were grown by vapour diffusion. The protein crystallizes in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 43.53, b = 121.30, c = 143.75 Å. Native data to a resolution of 2.25 Å were recorded at station PX 14.1 (Daresbury) from a single crystal. Preliminary analysis of these data suggests that the asymmetric unit contains four copies of the AbsC monomer, giving an estimated solvent content of 47.0%. AbsC belongs to the MarR family of proteins that mediate ligand-responsive transcriptional control.