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Sample records for metabolic products formed

  1. Metabolism of Zearalenone and Its Major Modified Forms in Pigs

    Sabina B. Binder

    2017-02-01

    Full Text Available The Fusarium mycotoxin zearalenone (ZEN can be conjugated with polar molecules, like sugars or sulfates, by plants and fungi. To date, the fate of these modified forms of ZEN has not yet been elucidated in animals. In order to investigate whether ZEN conjugates contribute to the total ZEN exposure of an individual, ZEN (10 µg/kg b.w. and equimolar amounts of two of its plant metabolites (ZEN-14-O-β-glucoside, ZEN-16-O-β-glucoside and of one fungal metabolite (ZEN-14-sulfate were orally administered to four pigs as a single bolus using a repeated measures design. The concentrations of ZEN, its modified forms and its mammalian metabolites ZEN-14-glucuronide, α-zearalenol (α-ZEL and α-ZEL-14-glucuronide in excreta were analyzed by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS based methods. The biological recovery of ZEN in urine was 26% ± 10%, the total biological recovery in excreta was 40% ± 8%. Intact ZEN-14-sulfate, ZEN-14-O-β-glucoside and ZEN-16-O-β-glucoside were neither detected in urine nor in feces. After ZEN-14-sulfate application, 19% ± 5% of the administered dose was recovered in urine. In feces, no ZEN metabolites were detected. The total biological recoveries of ZEN-14-O-β-glucoside and ZEN-16-O-β-glucoside in the form of their metabolites in urine were 19% ± 11% and 13% ± 7%, respectively. The total biological recoveries in urine and feces amounted to 48% ± 7% and 34 ± 3%. An explanation for the low biological recoveries could be extensive metabolization by intestinal bacteria to yet unknown metabolites. In summary, ZEN-14-sulfate, ZEN-14-O-β-glucoside, and ZEN-16-O-β-glucoside were completely hydrolyzed in the gastrointestinal tract of swine, thus contributing to the overall toxicity of ZEN.

  2. Patchoulol Production with Metabolically Engineered Corynebacterium glutamicum

    Nadja A. Henke

    2018-04-01

    Full Text Available Patchoulol is a sesquiterpene alcohol and an important natural product for the perfume industry. Corynebacterium glutamicum is the prominent host for the fermentative production of amino acids with an average annual production volume of ~6 million tons. Due to its robustness and well established large-scale fermentation, C. glutamicum has been engineered for the production of a number of value-added compounds including terpenoids. Both C40 and C50 carotenoids, including the industrially relevant astaxanthin, and short-chain terpenes such as the sesquiterpene valencene can be produced with this organism. In this study, systematic metabolic engineering enabled construction of a patchoulol producing C. glutamicum strain by applying the following strategies: (i construction of a farnesyl pyrophosphate-producing platform strain by combining genomic deletions with heterologous expression of ispA from Escherichia coli; (ii prevention of carotenoid-like byproduct formation; (iii overproduction of limiting enzymes from the 2-c-methyl-d-erythritol 4-phosphate (MEP-pathway to increase precursor supply; and (iv heterologous expression of the plant patchoulol synthase gene PcPS from Pogostemon cablin. Additionally, a proof of principle liter-scale fermentation with a two-phase organic overlay-culture medium system for terpenoid capture was performed. To the best of our knowledge, the patchoulol titers demonstrated here are the highest reported to date with up to 60 mg L−1 and volumetric productivities of up to 18 mg L−1 d−1.

  3. Metabolic engineering of cyanobacteria for the synthesis of commodity products

    Angermayr, S.A.; Gorchs Rovira, A.; Hellingwerf, K.J.

    2015-01-01

    Through metabolic engineering cyanobacteria can be employed in biotechnology. Combining the capacity for oxygenic photosynthesis and carbon fixation with an engineered metabolic pathway allows carbon-based product formation from CO2, light, and water directly. Such cyanobacterial 'cell factories'

  4. Ethylene production in relation to nitrogen metabolism in Saccharomyces cerevisiae.

    Johansson, Nina; Persson, Karl O; Quehl, Paul; Norbeck, Joakim; Larsson, Christer

    2014-11-01

    We have previously shown that ethylene production in Saccharomyces cerevisiae expressing the ethylene-forming enzyme (EFE) from Pseudomonas syringae is strongly influenced by variations in the mode of cultivation as well as the choice of nitrogen source. Here, we have studied the influence of nitrogen metabolism on the production of ethylene further. Using ammonium, glutamate, glutamate/arginine, and arginine as nitrogen sources, it was found that glutamate (with or without arginine) correlates with a high ethylene production, most likely linked to an observed increase in 2-oxoglutarate levels. Arginine as a sole nitrogen source caused a reduced ethylene production. A reduction of arginine levels, accomplished using an arginine auxotrophic ARG4-deletion strain in the presence of limiting amounts of arginine or through CAR1 overexpression, did however not correlate with an increased ethylene production. As expected, arginine was necessary for ethylene production as ethylene production in the ARG4-deletion strain ceased at the time when arginine was depleted. In conclusion, our data suggest that high levels of 2-oxoglutarate and a limited amount of arginine are required for successful ethylene production in yeast. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  5. Engineering strategy of yeast metabolism for higher alcohol production

    Shimizu Hiroshi

    2011-09-01

    Full Text Available Abstract Background While Saccharomyces cerevisiae is a promising host for cost-effective biorefinary processes due to its tolerance to various stresses during fermentation, the metabolically engineered S. cerevisiae strains exhibited rather limited production of higher alcohols than that of Escherichia coli. Since the structure of the central metabolism of S. cerevisiae is distinct from that of E. coli, there might be a problem in the structure of the central metabolism of S. cerevisiae. In this study, the potential production of higher alcohols by S. cerevisiae is compared to that of E. coli by employing metabolic simulation techniques. Based on the simulation results, novel metabolic engineering strategies for improving higher alcohol production by S. cerevisiae were investigated by in silico modifications of the metabolic models of S. cerevisiae. Results The metabolic simulations confirmed that the high production of butanols and propanols by the metabolically engineered E. coli strains is derived from the flexible behavior of their central metabolism. Reducing this flexibility by gene deletion is an effective strategy to restrict the metabolic states for producing target alcohols. In contrast, the lower yield using S. cerevisiae originates from the structurally limited flexibility of its central metabolism in which gene deletions severely reduced cell growth. Conclusions The metabolic simulation demonstrated that the poor productivity of S. cerevisiae was improved by the introduction of E. coli genes to compensate the structural difference. This suggested that gene supplementation is a promising strategy for the metabolic engineering of S. cerevisiae to produce higher alcohols which should be the next challenge for the synthetic bioengineering of S. cerevisiae for the efficient production of higher alcohols.

  6. Processes for production of alternative waste forms

    Ross, W.A.; Rusin, J.M.; McElroy, J.L.

    1979-01-01

    During the past 20 years, numerous waste forms and processes have been proposed for solidification of high-level radioactive wastes (HLW). The number has increased significantly during the past 3 to 4 years. At least five factors must be considered in selecting the waste form and process method: 1) processing flexibility, 2) waste loading, 3) canister size and stability, 4) waste form inertness and stability, and 5) processing complexity. This paper describes various waste form processes and operations, and a simple system is proposed for making comparisons. This system suggests that one goal for processes would be to reduce the number of process steps, thereby providing less complex processing systems

  7. Engineering yeast metabolism for production of fuels and chemicals

    Nielsen, Jens

    2016-01-01

    faster development of metabolically engineered strains that can be used for production of fuels and chemicals. The yeast Saccharomyces cerevisiae is widely used for production of fuels, chemicals, pharmaceuticals and materials. Through metabolic engineering of this yeast a number of novel industrial...... as for metabolic design. In this lecture it will be demonstrated how the Design-Build-Test cycle of metabolic engineering has allowed for development of yeast cell factories for production of a range of different fuels and chemicals. Some examples of different technologies will be presented together with examples......Metabolic engineering relies on the Design-Build-Test cycle. This cycle includes technologies like mathematical modeling of metabolism, genome editing and advanced tools for phenotypic characterization. In recent years there have been advances in several of these technologies, which has enabled...

  8. Novel sterol metabolic network of Trypanosoma brucei procyclic and bloodstream forms

    Nes, Craigen R.; Singha, Ujjal K.; Liu, Jialin; Ganapathy, Kulothungan; Villalta, Fernando; Waterman, Michael R.; Lepesheva, Galina I.; Chaudhuri, Minu; Nes, W. David

    2012-01-01

    Trypanosoma brucei is the protozoan parasite that causes African trypanosomiasis, a neglected disease of people and animals. Co-metabolite analysis, labelling studies using [methyl-2H3]-methionine and substrate/product specificities of the cloned 24-SMT (sterol C24-methyltransferase) and 14-SDM (sterol C14-demethylase) from T. brucei afforded an uncommon sterol metabolic network that proceeds from lanosterol and 31-norlanosterol to ETO [ergosta-5,7,25(27)-trien-3β-ol], 24-DTO [dimethyl ergosta-5,7,25(27)-trienol] and ergosterol [ergosta-5,7,22(23)-trienol]. To assess the possible carbon sources of ergosterol biosynthesis, specifically 13C-labelled specimens of lanosterol, acetate, leucine and glucose were administered to T. brucei and the 13C distributions found were in accord with the operation of the acetate–mevalonate pathway, with leucine as an alternative precursor, to ergostenols in either the insect or bloodstream form. In searching for metabolic signatures of procyclic cells, we observed that the 13C-labelling treatments induce fluctuations between the acetyl-CoA (mitochondrial) and sterol (cytosolic) synthetic pathways detected by the progressive increase in 13C-ergosterol production (control sterol synthesis that is further fluctuated in the cytosol, yielding distinct sterol profiles in relation to cell demands on growth. PMID:22176028

  9. Production of L-valine from metabolically engineered Corynebacterium glutamicum.

    Wang, Xiaoyuan; Zhang, Hailing; Quinn, Peter J

    2018-05-01

    L-Valine is one of the three branched-chain amino acids (valine, leucine, and isoleucine) essential for animal health and important in metabolism; therefore, it is widely added in the products of food, medicine, and feed. L-Valine is predominantly produced through microbial fermentation, and the production efficiency largely depends on the quality of microorganisms. In recent years, continuing efforts have been made in revealing the mechanisms and regulation of L-valine biosynthesis in Corynebacterium glutamicum, the most utilitarian bacterium for amino acid production. Metabolic engineering based on the metabolic biosynthesis and regulation of L-valine provides an effective alternative to the traditional breeding for strain development. Industrially competitive L-valine-producing C. glutamicum strains have been constructed by genetically defined metabolic engineering. This article reviews the global metabolic and regulatory networks responsible for L-valine biosynthesis, the molecular mechanisms of regulation, and the strategies employed in C. glutamicum strain engineering.

  10. Yeast metabolic engineering for hemicellulosic ethanol production

    Jennifer Van Vleet; Thomas W. Jeffries

    2009-01-01

    Efficient fermentation of hemicellulosic sugars is critical for the bioconversion of lignocellulosics to ethanol. Efficient sugar uptake through the heterologous expression of yeast and fungal xylose/glucose transporters can improve fermentation if other metabolic steps are not rate limiting. Rectification of cofactor imbalances through heterologous expression of...

  11. CAN THE END PRODUCTS OF ANAEROBIC METABOLISM ...

    Exercise, primarily powered by the shell adductor muscle, was mainly fueled by ... in seawater) transportation stress of up to 36 h at 7 and 10°C, clearly showed that ... at 10°C), indicating that aerobic metabolism is impaired at an early stage of ...

  12. Graphesis visual forms of knowledge production

    Drucker, Johanna

    2014-01-01

    In our current screen-saturated culture, we take in more information through visual means than at any point in history. The computers and smart phones that constantly flood us with images do more than simply convey information. They structure our relationship to information through graphical formats. Learning to interpret how visual forms not only present but produceknowledge, says Johanna Drucker, has become an essential contemporary skill. Graphesis provides a descriptive critical language for the analysis of graphical knowledge. In an interdisciplinary study fusing digital humanities with media studies and graphic design history, Drucker outlines the principles by which visual formats organize meaningful content. Among the most significant of these formats is the graphical user interface (GUI)--the dominant feature of the screens of nearly all consumer electronic devices. Because so much of our personal and professional lives is mediated through visual interfaces, it is important to start thinking critical...

  13. Production of amino acids - Genetic and metabolic engineering approaches.

    Lee, Jin-Ho; Wendisch, Volker F

    2017-12-01

    The biotechnological production of amino acids occurs at the million-ton scale and annually about 6milliontons of l-glutamate and l-lysine are produced by Escherichia coli and Corynebacterium glutamicum strains. l-glutamate and l-lysine production from starch hydrolysates and molasses is very efficient and access to alternative carbon sources and new products has been enabled by metabolic engineering. This review focusses on genetic and metabolic engineering of amino acid producing strains. In particular, rational approaches involving modulation of transcriptional regulators, regulons, and attenuators will be discussed. To address current limitations of metabolic engineering, this article gives insights on recent systems metabolic engineering approaches based on functional tools and method such as genome reduction, amino acid sensors based on transcriptional regulators and riboswitches, CRISPR interference, small regulatory RNAs, DNA scaffolding, and optogenetic control, and discusses future prospects. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Production of metal waste forms from spent fuel treatment

    Westphal, B.R.; Keiser, D.D.; Rigg, R.H.; Laug, D.V.

    1995-01-01

    Treatment of spent nuclear fuel at Argonne National Laboratory consists of a pyroprocessing scheme in which the development of suitable waste forms is being advanced. Of the two waste forms being proposed, metal and mineral, the production of the metal waste form utilizes induction melting to stabilize the waste product. Alloying of metallic nuclear materials by induction melting has long been an Argonne strength and thus, the transition to metallic waste processing seems compatible. A test program is being initiated to coalesce the production of the metal waste forms with current induction melting capabilities

  15. Probing the metabolic network in bloodstream-form Trypanosoma brucei using untargeted metabolomics with stable isotope labelled glucose.

    Darren J Creek

    2015-03-01

    Full Text Available Metabolomics coupled with heavy-atom isotope-labelled glucose has been used to probe the metabolic pathways active in cultured bloodstream form trypomastigotes of Trypanosoma brucei, a parasite responsible for human African trypanosomiasis. Glucose enters many branches of metabolism beyond glycolysis, which has been widely held to be the sole route of glucose metabolism. Whilst pyruvate is the major end-product of glucose catabolism, its transamination product, alanine, is also produced in significant quantities. The oxidative branch of the pentose phosphate pathway is operative, although the non-oxidative branch is not. Ribose 5-phosphate generated through this pathway distributes widely into nucleotide synthesis and other branches of metabolism. Acetate, derived from glucose, is found associated with a range of acetylated amino acids and, to a lesser extent, fatty acids; while labelled glycerol is found in many glycerophospholipids. Glucose also enters inositol and several sugar nucleotides that serve as precursors to macromolecule biosynthesis. Although a Krebs cycle is not operative, malate, fumarate and succinate, primarily labelled in three carbons, were present, indicating an origin from phosphoenolpyruvate via oxaloacetate. Interestingly, the enzyme responsible for conversion of phosphoenolpyruvate to oxaloacetate, phosphoenolpyruvate carboxykinase, was shown to be essential to the bloodstream form trypanosomes, as demonstrated by the lethal phenotype induced by RNAi-mediated downregulation of its expression. In addition, glucose derivatives enter pyrimidine biosynthesis via oxaloacetate as a precursor to aspartate and orotate.

  16. A regression-based Kansei engineering system based on form feature lines for product form design

    Yan Xiong

    2016-06-01

    Full Text Available When developing new products, it is important for a designer to understand users’ perceptions and develop product form with the corresponding perceptions. In order to establish the mapping between users’ perceptions and product design features effectively, in this study, we presented a regression-based Kansei engineering system based on form feature lines for product form design. First according to the characteristics of design concept representation, product form features–product form feature lines were defined. Second, Kansei words were chosen to describe image perceptions toward product samples. Then, multiple linear regression and support vector regression were used to construct the models, respectively, that predicted users’ image perceptions. Using mobile phones as experimental samples, Kansei prediction models were established based on the front view form feature lines of the samples. From the experimental results, these two predict models were of good adaptability. But in contrast to multiple linear regression, the predict performance of support vector regression model was better, and support vector regression is more suitable for form regression prediction. The results of the case showed that the proposed method provided an effective means for designers to manipulate product features as a whole, and it can optimize Kansei model and improve practical values.

  17. Engineering microorganisms to increase ethanol production by metabolic redirection

    Deng, Yu; Olson, Daniel G.; van Dijken, Johannes Pieter; Shaw, IV, Arthur J.; Argyros, Aaron; Barrett, Trisha; Caiazza, Nicky; Herring, Christopher D.; Rogers, Stephen R.; Agbogbo, Frank

    2017-10-31

    The present invention provides for the manipulation of carbon flux in a recombinant host cell to increase the formation of desirable products. The invention relates to cellulose-digesting organisms that have been genetically modified to allow the production of ethanol at a high yield by redirecting carbon flux at key steps of central metabolism.

  18. Metabolic engineering of biosynthetic pathway for production of renewable biofuels.

    Singh, Vijai; Mani, Indra; Chaudhary, Dharmendra Kumar; Dhar, Pawan Kumar

    2014-02-01

    Metabolic engineering is an important area of research that involves editing genetic networks to overproduce a certain substance by the cells. Using a combination of genetic, metabolic, and modeling methods, useful substances have been synthesized in the past at industrial scale and in a cost-effective manner. Currently, metabolic engineering is being used to produce sufficient, economical, and eco-friendly biofuels. In the recent past, a number of efforts have been made towards engineering biosynthetic pathways for large scale and efficient production of biofuels from biomass. Given the adoption of metabolic engineering approaches by the biofuel industry, this paper reviews various approaches towards the production and enhancement of renewable biofuels such as ethanol, butanol, isopropanol, hydrogen, and biodiesel. We have also identified specific areas where more work needs to be done in the future.

  19. Cyanobacterial metabolic engineering for biofuel and chemical production.

    Oliver, Neal J; Rabinovitch-Deere, Christine A; Carroll, Austin L; Nozzi, Nicole E; Case, Anna E; Atsumi, Shota

    2016-12-01

    Rising levels of atmospheric CO 2 are contributing to the global greenhouse effect. Large scale use of atmospheric CO 2 may be a sustainable and renewable means of chemical and liquid fuel production to mitigate global climate change. Photosynthetic organisms are an ideal platform for efficient, natural CO 2 conversion to a broad range of chemicals. Cyanobacteria are especially attractive for these purposes, due to their genetic malleability and relatively fast growth rate. Recent years have yielded a range of work in the metabolic engineering of cyanobacteria and have led to greater knowledge of the host metabolism. Understanding of endogenous and heterologous carbon regulation mechanisms leads to the expansion of productive capacity and chemical variety. This review discusses the recent progress in metabolic engineering of cyanobacteria for biofuel and bulk chemical production since 2014. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Metabolic engineering of microalgal based biofuel production: prospects and challenges

    Chiranjib eBanerjee

    2016-03-01

    Full Text Available The current scenario in renewable energy is focused on development of alternate and sustainable energy sources, amongst which microalgae stands as one of the promising feedstock for biofuel production. It is well known that microalgae generate much larger amounts of biofuels in a shorter time than other sources based on plant seeds. However, the greatest challenge in a transition to algae-based biofuel production is the various other complications involved in microalgal cultivation, its harvesting, concentration, drying and lipid extraction. Several green microalgae accumulate lipids, especially triacylglycerols (TAGs, which are main precursors in the production of lipid. The various aspects on metabolic pathway analysis of an oleaginous microalgae i.e. Chlamydomonas reinhardtii have elucidated some novel metabolically important genes and this enhances the lipid production in this microalgae. Adding to it, various other aspects in metabolic engineering using OptFlux and effectual bioprocess design also gives an interactive snapshot of enhancing lipid production which ultimately improvises the oil yield. This article reviews the current status of microalgal based technologies for biofuel production, bioreactor process design, flux analysis and it also provides various strategies to increase lipids accumulation via metabolic engineering.

  1. Central carbon metabolism influences cellulase production in Bacillus licheniformis.

    Wang, J; Liu, S; Li, Y; Wang, H; Xiao, S; Li, C; Liu, B

    2018-01-01

    Bacillus licheniformis that can produce cellulase including endo glucanase and glucosidase is an important industrial microbe for cellulose degradation. The purpose of this research was to assess the effect of endo glucanase gene bglC and glucosidase gene bglH on the central metabolic flux in B. licheniformis. bglC and bglH were knocked out using homologous recombination method, respectively, and the corresponding knockout strains were obtained for 13 C metabolic flux analysis. A significant change was observed in metabolic fluxes after 13 C metabolic flux ratio analysis. In both of the knockout strains, the increased fluxes of the pentose phosphate pathway and malic enzyme reaction enabled an elevated supply of NADPH which provided enough reducing power for the in vivo synthesis reactions. The fluxes through tricarboxylic acid cycle and anaplerotic reactions increased fast in the two knockout strains, which meant more energy generated. The changed fluxes in central carbon metabolism provided a holistic view of the physiological status in B. licheniformis and possible targets for further strain engineering. Cellulase is very important in the field of agriculture and bioenergy because of its degrading effect on cellulosic biomass. This study presented the effect of central carbon metabolism on cellulase production in Bacillus licheniformis. The study also provided a holistic view of the physiological status in B. licheniformis. The shifted metabolism provided a quantitative evaluation of the biosynthesis of cellulase and a priority ranked target list for further strain engineering. © 2017 The Society for Applied Microbiology.

  2. Dehydratase mediated 1-propanol production in metabolically engineered Escherichia coli

    Jain Rachit

    2011-11-01

    Full Text Available Abstract Background With the increasing consumption of fossil fuels, the question of meeting the global energy demand is of great importance in the near future. As an effective solution, production of higher alcohols from renewable sources by microorganisms has been proposed to address both energy crisis and environmental concerns. Higher alcohols contain more than two carbon atoms and have better physiochemical properties than ethanol as fuel substitutes. Results We designed a novel 1-propanol metabolic pathway by expanding the well-known 1,2-propanediol pathway with two more enzymatic steps catalyzed by a 1,2-propanediol dehydratase and an alcohol dehydrogenase. In order to engineer the pathway into E. coli, we evaluated the activities of eight different methylglyoxal synthases which play crucial roles in shunting carbon flux from glycolysis towards 1-propanol biosynthesis, as well as two secondary alcohol dehydrogenases of different origins that reduce both methylglyoxal and hydroxyacetone. It is evident from our results that the most active enzymes are the methylglyoxal synthase from Bacillus subtilis and the secondary alcohol dehydrogenase from Klebsiella pneumoniae, encoded by mgsA and budC respectively. With the expression of these two genes and the E. coli ydjG encoding methylglyoxal reductase, we achieved the production of 1,2-propanediol at 0.8 g/L in shake flask experiments. We then characterized the catalytic efficiency of three different diol dehydratases on 1,2-propanediol and identified the optimal one as the 1,2-propanediol dehydratase from Klebsiella oxytoca, encoded by the operon ppdABC. Co-expressing this enzyme with the above 1,2-propanediol pathway in wild type E. coli resulted in the production of 1-propanol at a titer of 0.25 g/L. Conclusions We have successfully established a new pathway for 1-propanol production by shunting the carbon flux from glycolysis. To our knowledge, it is the first time that this pathway has been

  3. Metabollic Engineering of Saccharomyces Cereviae a,omi acid metabolism for production of products of industrial interest

    Chen, Xiao

    -based processes. This study has focused on metabolic engineering of the amino acid metabolism in S. cerevisiae for production of two types of chemicals of industrial interest. The first chemical is δ-(L-α-aminoadipyl)–L-cysteinyl–D-valine (LLD-ACV). ACV belongs to non-ribosomal peptides (NRPs), which......Saccharomyces cerevisiae is widely used in microbial production of chemicals, metabolites and proteins, mainly because genetic manipulation of S. cerevisiae is relatively easy and experiences from its wide application in the existing industrial fermentations directly benefit new S. cerevisiae...

  4. Production of L-carnitine by secondary metabolism of bacteria

    Iborra José L

    2007-10-01

    Full Text Available Abstract The increasing commercial demand for L-carnitine has led to a multiplication of efforts to improve its production with bacteria. The use of different cell environments, such as growing, resting, permeabilized, dried, osmotically stressed, freely suspended and immobilized cells, to maintain enzymes sufficiently active for L-carnitine production is discussed in the text. The different cell states of enterobacteria, such as Escherichia coli and Proteus sp., which can be used to produce L-carnitine from crotonobetaine or D-carnitine as substrate, are analyzed. Moreover, the combined application of both bioprocess and metabolic engineering has allowed a deeper understanding of the main factors controlling the production process, such as energy depletion and the alteration of the acetyl-CoA/CoA ratio which are coupled to the end of the biotransformation. Furthermore, the profiles of key central metabolic activities such as the TCA cycle, the glyoxylate shunt and the acetate metabolism are seen to be closely interrelated and affect the biotransformation efficiency. Although genetically modified strains have been obtained, new strain improvement strategies are still needed, especially in Escherichia coli as a model organism for molecular biology studies. This review aims to summarize and update the state of the art in L-carnitine production using E. coli and Proteus sp, emphasizing the importance of proper reactor design and operation strategies, together with metabolic engineering aspects and the need for feed-back between wet and in silico work to optimize this biotransformation.

  5. Hydrogen production and metabolic flux analysis of metabolically engineered Escherichia coli strains

    Kim, Seohyoung; Seol, Eunhee; Park, Sunghoon [Department of Chemical and Biochemical Engineering, Pusan National University, Busan 609-735 (Korea); Oh, You-Kwan [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-543 (Korea); Wang, G.Y. [Department of Oceanography, University of Hawaii at Manoa Honolulu, HI 96822 (United States)

    2009-09-15

    Escherichia coli can produce H{sub 2} from glucose via formate hydrogen lyase (FHL). In order to improve the H{sub 2} production rate and yield, metabolically engineered E. coli strains, which included pathway alterations in their H{sub 2} production and central carbon metabolism, were developed and characterized by batch experiments and metabolic flux analysis. Deletion of hycA, a negative regulator for FHL, resulted in twofold increase of FHL activity. Deletion of two uptake hydrogenases (1 (hya) and hydrogenase 2 (hyb)) increased H{sub 2} production yield from 1.20 mol/mol glucose to 1.48 mol/mol glucose. Deletion of lactate dehydrogenase (ldhA) and fumarate reductase (frdAB) further improved the H{sub 2} yield; 1.80 mol/mol glucose under high H{sub 2} pressure or 2.11 mol/mol glucose under reduced H{sub 2} pressure. Several batch experiments at varying concentrations of glucose (2.5-10 g/L) and yeast extract (0.3 or 3.0 g/L) were conducted for the strain containing all these genetic alternations, and their carbon and energy balances were analyzed. The metabolic flux analysis revealed that deletion of ldhA and frdAB directed most of the carbons from glucose to the glycolytic pathway leading to H{sub 2} production by FHL, not to the pentose phosphate pathway. (author)

  6. Advances in Metabolic Engineering of Cyanobacteria for Photosynthetic Biochemical Production

    Lai, Martin C.; Lan, Ethan I.

    2015-01-01

    Engineering cyanobacteria into photosynthetic microbial cell factories for the production of biochemicals and biofuels is a promising approach toward sustainability. Cyanobacteria naturally grow on light and carbon dioxide, bypassing the need of fermentable plant biomass and arable land. By tapping into the central metabolism and rerouting carbon flux towards desirable compound production, cyanobacteria are engineered to directly convert CO2 into various chemicals. This review discusses the d...

  7. Metabolic engineering of Escherichia coli for the production of riboflavin

    Lin, Zhenquan; Xu, Zhibo; Li, Yifan; Wang, Zhiwen; Chen, Tao; Zhao, Xueming

    2014-01-01

    Background Riboflavin (vitamin B2), the precursor of the flavin cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), is used commercially as an animal feed supplement and food colorant. E. coli is a robust host for various genetic manipulations and has been employed for efficient production of biofuels, polymers, amino acids, and bulk chemicals. Thus, the aim of this study was to understand the metabolic capacity of E. coli for the riboflavin production by modification...

  8. Metabolic heat production by human and animal populations in cities

    Stewart, Iain D.; Kennedy, Chris A.

    2017-07-01

    Anthropogenic heating from building energy use, vehicle fuel consumption, and human metabolism is a key term in the urban energy budget equation. Heating from human metabolism, however, is often excluded from urban energy budgets because it is widely observed to be negligible. Few reports for low-latitude cities are available to support this observation, and no reports exist on the contribution of domestic animals to urban heat budgets. To provide a more comprehensive view of metabolic heating in cities, we quantified all terms of the anthropogenic heat budget at metropolitan scale for the world's 26 largest cities, using a top-down statistical approach. Results show that metabolic heat release from human populations in mid-latitude cities (e.g. London, Tokyo, New York) accounts for 4-8% of annual anthropogenic heating, compared to 10-45% in high-density tropical cities (e.g. Cairo, Dhaka, Kolkata). Heat release from animal populations amounts to <1% of anthropogenic heating in all cities. Heat flux density from human and animal metabolism combined is highest in Mumbai—the world's most densely populated megacity—at 6.5 W m-2, surpassing heat production by electricity use in buildings (5.8 W m-2) and fuel combustion in vehicles (3.9 W m-2). These findings, along with recent output from global climate models, suggest that in the world's largest and most crowded cities, heat emissions from human metabolism alone can force measurable change in mean annual temperature at regional scale.

  9. Methods for forming complex oxidation reaction products including superconducting articles

    Rapp, R.A.; Urquhart, A.W.; Nagelberg, A.S.; Newkirk, M.S.

    1992-01-01

    This patent describes a method for producing a superconducting complex oxidation reaction product of two or more metals in an oxidized state. It comprises positioning at least one parent metal source comprising one of the metals adjacent to a permeable mass comprising at least one metal-containing compound capable of reaction to form the complex oxidation reaction product in step below, the metal component of the at least one metal-containing compound comprising at least a second of the two or more metals, and orienting the parent metal source and the permeable mass relative to each other so that formation of the complex oxidation reaction product will occur in a direction towards and into the permeable mass; and heating the parent metal source in the presence of an oxidant to a temperature region above its melting point to form a body of molten parent metal to permit infiltration and reaction of the molten parent metal into the permeable mass and with the oxidant and the at least one metal-containing compound to form the complex oxidation reaction product, and progressively drawing the molten parent metal source through the complex oxidation reaction product towards the oxidant and towards and into the adjacent permeable mass so that fresh complex oxidation reaction product continues to form within the permeable mass; and recovering the resulting complex oxidation reaction product

  10. Metabolic Engineering of Microorganisms for the Production of Higher Alcohols

    Choi, Yong Jun; Lee, Joungmin; Jang, Yu-Sin

    2014-01-01

    ABSTRACT Due to the increasing concerns about limited fossil resources and environmental problems, there has been much interest in developing biofuels from renewable biomass. Ethanol is currently used as a major biofuel, as it can be easily produced by existing fermentation technology, but it is not the best biofuel due to its low energy density, high vapor pressure, hygroscopy, and incompatibility with current infrastructure. Higher alcohols, including 1-propanol, 1-butanol, isobutanol, 2-methyl-1-butanol, and 3-methyl-1-butanol, which possess fuel properties more similar to those of petroleum-based fuel, have attracted particular interest as alternatives to ethanol. Since microorganisms isolated from nature do not allow production of these alcohols at high enough efficiencies, metabolic engineering has been employed to enhance their production. Here, we review recent advances in metabolic engineering of microorganisms for the production of higher alcohols. PMID:25182323

  11. Production of vanillin by metabolically engineered Escherichia coli.

    Yoon, Sang-Hwal; Li, Cui; Kim, Ju-Eun; Lee, Sook-Hee; Yoon, Ji-Young; Choi, Myung-Suk; Seo, Weon-Taek; Yang, Jae-Kyung; Kim, Jae-Yeon; Kim, Seon-Won

    2005-11-01

    E. coli was metabolically engineered to produce vanillin by expression of the fcs and ech genes from Amycolatopsis sp. encoding feruloyl-CoA synthetase and enoyl-CoA hydratase/aldolase, respectively. Vanillin production was optimized by leaky expression of the genes, under the IPTG-inducible trc promoter, in complex 2YT medium. Supplementation with glucose, fructose, galactose, arabinose or glycerol severely decreased vanillin production. The highest vanillin production of 1.1 g l(-1) was obtained with cultivation for 48 h in 2YT medium with 0.2% (w/v) ferulate, without IPTG and no supplementation of carbon sources.

  12. Improving lactate metabolism in an intensified CHO culture process: productivity and product quality considerations.

    Xu, Sen; Hoshan, Linda; Chen, Hao

    2016-11-01

    In this study, we discussed the development and optimization of an intensified CHO culture process, highlighting medium and control strategies to improve lactate metabolism. A few strategies, including supplementing glucose with other sugars (fructose, maltose, and galactose), controlling glucose level at Productivity and product quality attributes differences between batch, fed-batch, and concentrated fed-batch cultures were discussed. The importance of process and cell metabolism understanding when adapting the existing process to a new operational mode was demonstrated in the study.

  13. Biobased organic acids production by metabolically engineered microorganisms

    Chen, Yun; Nielsen, Jens

    2016-01-01

    Bio-based production of organic acids via microbial fermentation has been traditionally used in food industry. With the recent desire to develop more sustainable bioprocesses for production of fuels, chemicals and materials, the market for microbial production of organic acids has been further...... expanded as organic acids constitute a key group among top building block chemicals that can be produced from renewable resources. Here we review the current status for production of citric acid and lactic acid, and we highlight the use of modern metabolic engineering technologies to develop high...... performance microbes for production of succinic acid and 3-hydroxypropionic acid. Also, the key limitations and challenges in microbial organic acids production are discussed...

  14. How language production shapes language form and comprehension

    Maryellen C MacDonald

    2013-04-01

    Full Text Available Language production processes can provide insight into how language comprehension works and language typology—why languages tend to have certain characteristics more often than others. Drawing on work in memory retrieval, motor planning, and serial order in action planning, the Production-Distribution-Comprehension (PDC account links work in the fields of language production, typology, and comprehension: 1 faced with substantial computational burdens of planning and producing utterances, language producers implicitly follow three biases in utterance planning that promote word order choices that reduce these burdens, thereby improving production fluency. 2 These choices, repeated over many utterances and individuals, shape the distributions of utterance forms in language. The claim that language form stems in large degree from producers’ attempts to mitigate utterance planning difficulty is contrasted with alternative accounts in which form is driven by language use more broadly, language acquisition processes, or producers’ attempts to create language forms that are easily understood by comprehenders. 3 Language perceivers implicitly learn the statistical regularities in their linguistic input, and they use this prior experience to guide comprehension of subsequent language. In particular, they learn to predict the sequential structure of linguistic signals, based on the statistics of previously-encountered input. Thus key aspects of comprehension behavior are tied to lexico-syntactic statistics in the language, which in turn derive from utterance planning biases promoting production of comparatively easy utterance forms over more difficult ones. This approach contrasts with classic theories in which comprehension behaviors are attributed to innate design features of the language comprehension system and associated working memory. The PDC instead links basic features of comprehension to a different source: production processes that shape

  15. Reactions of newly formed fission products in the gas phase

    Strickert, R.G.

    1976-01-01

    A dynamic gas-flow system was constructed which stopped fission products in the gas phase and rapidly separated (in less than 2 sec) volatile compounds from non-volatile ones. The filter assembly designed and used was shown to stop essentially all non-volatile fission products. Between 5 percent and 20 percent of tellurium fission-product isotopes reacted with several hydrocarbon gases to form volatile compounds, which passed through the filter. With carbon monoxide gas, volatile tellurium compound(s) (probably TeCO) were also formed with similar efficiencies. The upper limits for the yields of volatile compounds formed between CO and tin and antimony fission products were shown to be less than 0.3 percent, so tellurium nuclides, not their precursors, reacted with CO. It was found that CO reacted preferentially with independently produced tellurium atoms; the reaction efficiency of beta-produced atoms was only 27 +- 3 percent of that of the independently formed atoms. The selectivity, which was independent of the over-all reaction efficiency, was shown to be due to reaction of independently formed atoms in the gas phase. The gas phase reactions are believed to occur mainly at thermal energies because of the independence of the yield upon argon moderator mole-fraction (up to 80 percent). It was shown in some experiments that about one-half of the TeCO decomposed in passing through a filter and that an appreciable fraction (approximately 20 percent) of the tellurium atoms deposited on the filter reacted agin with CO. Other tellurium atoms on the filter surface (those formed by beta decay and those formed independently but not reacting in the gas phase) also reacted with CO, but probably somewhat less efficiently than atoms formed by TeCO decomposition. No evidence was found for formation of TeCO as a direct result of beta-decay

  16. Overexpression of a water-forming NADH oxidase improves the metabolism and stress tolerance of Saccharmyces cerevisiae in aerobic fermenation

    Xinchi Shi

    2016-09-01

    Full Text Available Redox homeostasis is fundamental to the maintenance of metabolism. Redox imbalance can cause oxidative stress, which affects metabolism and growth. Water-forming NADH oxidase regulates the redox balance by oxidizing cytosolic NADH to NAD+, which relieves cytosolic NADH accumulation through rapid glucose consumption in Saccharomyces cerevisiae, thus decreasing the production of the byproduct glycerol in industrial ethanol production. Here, we studied the effects of overexpression of a water-forming NADH oxidase from Lactococcus lactis on the stress response of S. cerevisiae in aerobic batch fermentation, and we constructed an interaction network of transcriptional regulation and metabolic networks to study the effects of and mechanisms underlying NADH oxidase regulation. The oxidase-overexpressing strain (NOX showed increased glucose consumption, growth, and ethanol production, while glycerol production was remarkably lower. Glucose was exhausted by NOX at 26 h, while 18.92 ± 0.94 g/L residual glucose was left in the fermentation broth of the control strain (CON at this time point. At 29.5 h, the ethanol concentration for NOX peaked at 35.25 ± 1.76 g/L, which was 14.37 % higher than that for CON (30.82 ± 1.54 g/L. Gene expression involved in the synthesis of thiamine, which is associated with stress responses in various organisms, was increased in NOX. The transcription factor HAP4 was significantly upregulated in NOX at the late-exponential phase, indicating a diauxic shift in response to starvation. The apoptosis-inducing factor Nuc1 was downregulated while the transcription factor Sok2, which regulates the production of the small signaling molecule ammonia, was upregulated at the late-exponential phase, benefiting young cells on the rim. Reactive oxygen species production was decreased by 10% in NOX, supporting a decrease in apoptosis. The HOG pathway was not activated, although the osmotic stress was truly higher, indicating improved

  17. Equilibrium Temperature Profiles within Fission Product Waste Forms

    Kaminski, Michael D. [Argonne National Lab. (ANL), Argonne, IL (United States)

    2016-10-01

    We studied waste form strategies for advanced fuel cycle schemes. Several options were considered for three waste streams with the following fission products: cesium and strontium, transition metals, and lanthanides. These three waste streams may be combined or disposed separately. The decay of several isotopes will generate heat that must be accommodated by the waste form, and this heat will affect the waste loadings. To help make an informed decision on the best option, we present computational data on the equilibrium temperature of glass waste forms containing a combination of these three streams.

  18. Calorimetric study on human erythrocyte glycolysis. Heat production in various metabolic conditions.

    Minakami, S; de Verdier, C H

    1976-06-01

    The heat production of human erythrocytes was measured on a flow microcalorimeter with simultaneous analyses of lactate and other metabolites. The heat production connected with the lactate formation was about 17 kcal (71 kJ) per mol lactate formed which corresponded to the sum of heat production due to the formation of lactate from glucose and the heat production due to neutralization. The heat production rate increased as the pH of the suspension increased, corresponding to the increase in lactate formation. Glycolytic inhibitors such as fluoride and monoiodoacetate caused a decrease in the rate of heat production, whereas arsenate induced a large transient increase in heat production associated with a transient increase in lactate formation. Decrease in pyruvate concentration was usually associated with increase in heat production, although the decreased pyruvate concentration was coupled with formation of 2,3-bisphosphoglycerate. When inosine, dihydroxyacetone or D-glyceraldehyde was used as a substrate, an increase in the heat production rate was observed. Addition of methylene blue caused an oxygen uptake which was accompanied by a remarkable increase in heat production rate corresponding to about 160 kcal (670 kJ) per mol oxygen consumed. The value for heat production in red cells in the above-mentioned metabolic conditions was considered in relation to earlier known data on free energy and enthalpy changes of the different metabolic steps in the glycolytic pathway.

  19. The Metabolic Properties of the Fission Products and Actinide Elements

    Hamilton M.D., J.G.

    1948-03-01

    An investigation of the assimilation, distribution, retention, an excretion of the fission products and actinide elements in the rat has been conducted at the Crocker Radiation Laboratory, University of California, Berkeley, California. These studies were initiated October 15, 1942, and are continuing at the present time. An extensive survey has been made of the metabolism of twenty-two different radio elements in the rat.

  20. Adjuvant activity of peptidoglycan monomer and its metabolic products.

    Halassy, Beata; Krstanović, Marina; Frkanec, Ruza; Tomasić, Jelka

    2003-02-14

    Peptidoglycan monomer (PGM) is a natural compound of bacterial origin. It is a non-toxic, non-pyrogenic, water-soluble immunostimulator potentiating humoral immune response to ovalbumin (OVA) in mice. It is fast degraded and its metabolic products-the pentapeptide (PP) and the disaccharide (DS)-are excreted from the mammalian organism upon parenteral administration. The present study investigates: (a). whether PGM could influence the long-living memory generation; (b). whether metabolic products retain adjuvant properties of the parent compound and contribute to its adjuvanticity. We report now that mice immunised twice with OVA+PGM had significantly higher anti-OVA IgG levels upon challenge with antigen alone 6 months later in comparison to control group immunised with OVA only. PP and DS were prepared enzymatically in vitro as apyrogenic and chemically pure compounds. When mice were immunised with OVA plus PP and DS, respectively, the level of anti-OVA IgGs in sera was not higher than in mice immunised with OVA alone, while PGM raised the level of specific antibodies. Results implicate that the adjuvant active molecule, capable of enhancing long-living memory generation, is PGM itself, and none of its metabolic products.

  1. Metabolic heat production by human and animal populations in cities.

    Stewart, Iain D; Kennedy, Chris A

    2017-07-01

    Anthropogenic heating from building energy use, vehicle fuel consumption, and human metabolism is a key term in the urban energy budget equation. Heating from human metabolism, however, is often excluded from urban energy budgets because it is widely observed to be negligible. Few reports for low-latitude cities are available to support this observation, and no reports exist on the contribution of domestic animals to urban heat budgets. To provide a more comprehensive view of metabolic heating in cities, we quantified all terms of the anthropogenic heat budget at metropolitan scale for the world's 26 largest cities, using a top-down statistical approach. Results show that metabolic heat release from human populations in mid-latitude cities (e.g. London, Tokyo, New York) accounts for 4-8% of annual anthropogenic heating, compared to 10-45% in high-density tropical cities (e.g. Cairo, Dhaka, Kolkata). Heat release from animal populations amounts to heating in all cities. Heat flux density from human and animal metabolism combined is highest in Mumbai-the world's most densely populated megacity-at 6.5 W m -2 , surpassing heat production by electricity use in buildings (5.8 W m -2 ) and fuel combustion in vehicles (3.9 W m -2 ). These findings, along with recent output from global climate models, suggest that in the world's largest and most crowded cities, heat emissions from human metabolism alone can force measurable change in mean annual temperature at regional scale.

  2. Automobile sheet metal part production with incremental sheet forming

    İsmail DURGUN

    2016-02-01

    Full Text Available Nowadays, effect of global warming is increasing drastically so it leads to increased interest on energy efficiency and sustainable production methods. As a result of adverse conditions, national and international project platforms, OEMs (Original Equipment Manufacturers, SMEs (Small and Mid-size Manufacturers perform many studies or improve existing methodologies in scope of advanced manufacturing techniques. In this study, advanced manufacturing and sustainable production method "Incremental Sheet Metal Forming (ISF" was used for sheet metal forming process. A vehicle fender was manufactured with or without die by using different toolpath strategies and die sets. At the end of the study, Results have been investigated under the influence of method and parameters used.Keywords: Template incremental sheet metal, Metal forming

  3. Correlation of Diffusion and Metabolic Alterations in Different Clinical Forms of Multiple Sclerosis

    Hannoun, Salem; Bagory, Matthieu; Durand-Dubief, Francoise; Ibarrola, Danielle; Comte, Jean-Christophe; Confavreux, Christian; Cotton, Francois; Sappey-Marinier, Dominique

    2012-01-01

    Diffusion tensor imaging (DTI) and MR spectroscopic imaging (MRSI) provide greater sensitivity than conventional MRI to detect diffuse alterations in normal appearing white matter (NAWM) of Multiple Sclerosis (MS) patients with different clinical forms. Therefore, the goal of this study is to combine DTI and MRSI measurements to analyze the relation between diffusion and metabolic markers, T2-weighted lesion load (T2-LL) and the patients clinical status. The sensitivity and specificity of both methods were then compared in terms of MS clinical forms differentiation. MR examination was performed on 71 MS patients (27 relapsing remitting (RR), 26 secondary progressive (SP) and 18 primary progressive (PP)) and 24 control subjects. DTI and MRSI measurements were obtained from two identical regions of interest selected in left and right centrum semioval (CSO) WM. DTI metrics and metabolic contents were significantly altered in MS patients with the exception of N-acetyl-aspartate (NAA) and NAA/Choline (Cho) ratio in RR patients. Significant correlations were observed between diffusion and metabolic measures to various degrees in every MS patients group. Most DTI metrics were significantly correlated with the T2-LL while only NAA/Cr ratio was correlated in RR patients. A comparison analysis of MR methods efficiency demonstrated a better sensitivity/specificity of DTI over MRSI. Nevertheless, NAA/Cr ratio could distinguish all MS and SP patients groups from controls, while NAA/Cho ratio differentiated PP patients from controls. This study demonstrated that diffusivity changes related to microstructural alterations were correlated with metabolic changes and provided a better sensitivity to detect early changes, particularly in RR patients who are more subject to inflammatory processes. In contrast, the better specificity of metabolic ratios to detect axonal damage and demyelination may provide a better index for identification of PP patients. PMID:22479330

  4. Estimating product-to-product variations in metal forming using force measurements

    Havinga, Gosse Tjipke; Van Den Boogaard, Ton

    2017-01-01

    The limits of production accuracy of metal forming processes can be stretched by the development of control systems for compensation of product-to-product variations. Such systems require the use of measurements from each semi-finished product. These measurements must be used to estimate the final

  5. Production of biopharmaceutical proteins by yeast: Advances through metabolic engineering

    Nielsen, Jens

    2013-01-01

    Production of recombinant proteins for use as pharmaceuticals, so-called biopharmaceuticals, is a multi-billion dollar industry. Many different cell factories are used for the production of biopharmaceuticals, but the yeast Saccharomyces cerevisiae is an important cell factory as it is used for p...... production. The involvement of directed metabolic engineering through the integration of tools from genetic engineering, systems biology and mathematical modeling, is also discussed....... by yeast are human serum albumin, hepatitis vaccines and virus like particles used for vaccination against human papillomavirus. Here is given a brief overview of biopharmaceutical production by yeast and it is discussed how the secretory pathway can be engineered to ensure more efficient protein...

  6. Metabolic engineering of Candida glabrata for diacetyl production.

    Xiang Gao

    Full Text Available In this study, Candida glabrata, an efficient pyruvate-producing strain, was metabolically engineered for the production of the food ingredient diacetyl. A diacetyl biosynthetic pathway was reconstructed based on genetic modifications and medium optimization. The former included (i channeling carbon flux into the diacetyl biosynthetic pathway by amplification of acetolactate synthase, (ii elimination of the branched pathway of α-acetolactate by deleting the ILV5 gene, and (iii restriction of diacetyl degradation by deleting the BDH gene. The resultant strain showed an almost 1∶1 co-production of α-acetolactate and diacetyl (0.95 g L(-1. Furthermore, addition of Fe3+ to the medium enhanced the conversion of α-acetolactate to diacetyl and resulted in a two-fold increase in diacetyl production (2.1 g L(-1. In addition, increased carbon flux was further channeled into diacetyl biosynthetic pathway and a titer of 4.7 g L(-1 of diacetyl was achieved by altering the vitamin level in the flask culture. Thus, this study illustrates that C. glabrata could be tailored as an attractive platform for enhanced biosynthesis of beneficial products from pyruvate by metabolic engineering strategies.

  7. Microbial production of antioxidant food ingredients via metabolic engineering.

    Lin, Yuheng; Jain, Rachit; Yan, Yajun

    2014-04-01

    Antioxidants are biological molecules with the ability to protect vital metabolites from harmful oxidation. Due to this fascinating role, their beneficial effects on human health are of paramount importance. Traditional approaches using solvent-based extraction from food/non-food sources and chemical synthesis are often expensive, exhaustive, and detrimental to the environment. With the advent of metabolic engineering tools, the successful reconstitution of heterologous pathways in Escherichia coli and other microorganisms provides a more exciting and amenable alternative to meet the increasing demand of natural antioxidants. In this review, we elucidate the recent progress in metabolic engineering efforts for the microbial production of antioxidant food ingredients - polyphenols, carotenoids, and antioxidant vitamins. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. A metabolic derivation of tritium transfer factors in animal products

    Galeriu, D.; Melintescu, A.; Crout, N. M. J.; Bersford, N. A.; Peterson, S. R.; Hess, M. van

    2001-01-01

    Tritium is a potentially important environmental contaminant arising from the nuclear industry. Because tritium is an isotope of hydrogen, its behaviour in the environment is controlled by the behaviour of hydrogen. Chronic releases of tritium to the atmosphere, in particular, will result in tritium-to-hydrogen (T/H) ratios in plants and animals that are more or less in equilibrium with T/H ratios in the air moisture. Tritium is thus a potentially important contaminant of plant and animal food products. The transfer of tritium from air moisture to plants is quite well understood. In contrast, although a number of regulatory agencies have published transfer coefficient values for diet tritium transfer for a limited number of animal products, a fresh evaluation of these transfers needs to be made In this paper we present an approach for the derivation of tritium transfer coefficients which is based on the metabolism of hydrogen in animals in conjunction with experimental data on tritium transfer. The derived transfer coefficients separately account for transfer to and from free (i.e. water) and organically bound tritium. The predicted transfer coefficients are compared to available data independent of model development. Agreement is good, with the exception of the transfer coefficient for transfer from tritiated water to organically bound tritium in ruminants, which may be attributable to the particular characteristics of ruminant digestion. We show that transfer coefficients will vary in response to the metabolic status of an animal (e.g. stage of lactation, digestibility of diet, etc.) and that the use of a single transfer coefficient from diet to animal product is not appropriate for tritium. It is possible to derive concentration ratio values which relate the concentration of tritiated water and organically bound tritium in an animal product to the corresponding concentrations in the animals diet. These concentration ratios are shown to be less subject to

  9. Production Equipment and Processes for Bulk Formed Micro Components

    Paldan, Nikolas Aulin; Arentoft, Mogens; Eriksen, Rasmus Solmer

    2007-01-01

    Manufacturing techniques for production of small precise metallic parts has gained interest during recent years, an interest led by an industrial demand for components for integrated products like mobile phones, personal digital assistants (PDAs), mp3-players and in the future for spare parts...... has been developed and used to form a number of industrial micro parts in aluminium and silver, with ongoing work on forming of titanium. Manufacture of billets by cropping has been examined using a simple test rig and an automatic cropping device has been designed, manufactured and tested....... for the human body. Micro components have also found several applications within the medical, audiological and dental industry, applications that impose increased demands for biocompatible and corrosion-resistant materials and cleanness. So far these micro components have mainly been manufactured by traditional...

  10. Programming Saposin-Mediated Compensatory Metabolic Sinks for Enhanced Ubiquinone Production.

    Xu, Wen; Yuan, Jifeng; Yang, Shuiyun; Ching, Chi-Bun; Liu, Jiankang

    2016-12-16

    Microbial synthesis of ubiquinone by fermentation processes has been emerging in recent years. However, as ubiquinone is a primary metabolite that is tightly regulated by the host central metabolism, tweaking the individual pathway components could only result in a marginal improvement on the ubiquinone production. Given that ubiquinone is stored in the lipid bilayer, we hypothesized that introducing additional metabolic sink for storing ubiquinone might improve the CoQ 10 production. As human lipid binding/transfer protein saposin B (hSapB) was reported to extract ubiquinone from the lipid bilayer and form the water-soluble complex, hSapB was chosen to build a compensatory metabolic sink for the ubiquinone storage. As a proof-of-concept, hSapB-mediated metabolic sink systems were devised and systematically investigated in the model organism of Escherichia coli. The hSapB-mediated periplasmic sink resulted in more than 200% improvement of CoQ 8 over the wild type strain. Further investigation revealed that hSapB-mediated sink systems could also improve the CoQ 10 production in a CoQ 10 -hyperproducing E. coli strain obtained by a modular pathway rewiring approach. As the design principles and the engineering strategies reported here are generalizable to other microbes, compensatory sink systems will be a method of significant interest to the synthetic biology community.

  11. Metabolism of pharmaceutical and personal care products by carrot cell cultures

    Wu, Xiaoqin; Fu, Qiuguo; Gan, Jay

    2016-01-01

    With the increasing use of treated wastewater and biosolids in agriculture, residues of pharmaceutical and personal care products (PPCPs) in these reused resources may contaminate food produce via plant uptake, constituting a route for human exposure. Although various PPCPs have been reported to be taken up by plants in laboratories or under field conditions, at present little information is available on their metabolism in plants. In this study, we applied carrot cell cultures to investigate the plant metabolism of PPCPs. Five phase I metabolites of carbamazepine were identified and the potential metabolism pathways of carbamazepine were proposed. We also used the carrot cell cultures as a rapid screening tool to initially assess the metabolism potentials of 18 PPCPs. Eleven PPCPs, including acetaminophen, caffeine, meprobamate, primidone, atenolol, trimethoprim, DEET, carbamazepine, dilantin, diazepam, and triclocarban, were found to be recalcitrant to metabolism. The other 7 PPCPs, including triclosan, naproxen, diclofenac, ibuprofen, gemfibrozil, sulfamethoxazole, and atorvastatin, displayed rapid metabolism, with 0.4–47.3% remaining in the culture at the end of the experiment. Further investigation using glycosidase hydrolysis showed that 1.3–20.6% of initially spiked naproxen, diclofenac, ibuprofen, and gemfibrozil were transformed into glycoside conjugates. Results from this study showed that plant cell cultures may be a useful tool for initially exploring the potential metabolites of PPCPs in plants as well as for rapidly screening the metabolism potentials of a variety of PPCPs or other emerging contaminants, and therefore may be used for prioritizing compounds for further comprehensive evaluations. - Highlights: • Five phase I metabolites of carbamazepine were identified in carrot cell cultures. • The metabolism potentials of 18 PPCPs were evaluated using carrot cell cultures. • Four PPCPs may partially form glycoside conjugates as phase II

  12. Production of sodalite waste forms by addition of glass

    Pereira, C.

    1995-01-01

    Spent nuclear fuel can be treated in a molten salt electrorefiner for conversion into metal and mineral waste forms for geologic disposal. Sodalite is one of the mineral waste forms under study. Fission products in the molten salt are ion-exchanged into zeolite A, which is converted to sodalite and consolidated. Sodalite can be formed directly from mixtures of salt and zeolite A at temperatures above 975 K; however, nepheline is usually produced as a secondary phase. Addition of small amounts of glass frit to the mixture reduced nepheline formation significantly. Loss of fission products was not observed for reaction below 1000 K. Hot-pressing of the sodalite powders yielded dense pellets (∼2.3 g/cm 3 ) without any loss of fission product species. Normalized release rates were below 1 g/m 2 ·day for pre-washed samples in 28-day leach tests based on standard MCC-1 tests but increased with the presence of free salt on the sodalite

  13. Metabolic inhibitors as stimulating factors for citric acid production

    Adham, N.Z.; Ahmed, E.M.; Refai, H.A.E.

    2008-01-01

    The effect of some metabolic inhibitors on citric acid (CA) production by Aspergillus niger in cane molasses medium was investigated. Addition of 0.01-0.1 mM iodoacetic acid and sodium arsenate, 0.05-1.0 mM sodium malonate, 0.01 mM sodium azide, 0.01-0.05 mM sodium fluoride, 0.1-1.0 mM EDTA stimulated CA production (5-49%). Higher concentrations (10 mM) of iodoacetic acid, sodium malonate and 0.5 mM sodium azide caused a complete inhibition of fungal growth, Iodoacetic acid, sodium arsenate and sodium fluoride (0.2 mM) caused a remarkable inhibition of CA production. The implications of those preliminary functions was discussed. (author)

  14. Enhancing microbial production of biofuels by expanding microbial metabolic pathways.

    Yu, Ping; Chen, Xingge; Li, Peng

    2017-09-01

    Fatty acid, isoprenoid, and alcohol pathways have been successfully engineered to produce biofuels. By introducing three genes, atfA, adhE, and pdc, into Escherichia coli to expand fatty acid pathway, up to 1.28 g/L of fatty acid ethyl esters can be achieved. The isoprenoid pathway can be expanded to produce bisabolene with a high titer of 900 mg/L in Saccharomyces cerevisiae. Short- and long-chain alcohols can also be effectively biosynthesized by extending the carbon chain of ketoacids with an engineered "+1" alcohol pathway. Thus, it can be concluded that expanding microbial metabolic pathways has enormous potential for enhancing microbial production of biofuels for future industrial applications. However, some major challenges for microbial production of biofuels should be overcome to compete with traditional fossil fuels: lowering production costs, reducing the time required to construct genetic elements and to increase their predictability and reliability, and creating reusable parts with useful and predictable behavior. To address these challenges, several aspects should be further considered in future: mining and transformation of genetic elements related to metabolic pathways, assembling biofuel elements and coordinating their functions, enhancing the tolerance of host cells to biofuels, and creating modular subpathways that can be easily interconnected. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

  15. Metabolic engineering of yeast for lignocellulosic biofuel production.

    Jin, Yong-Su; Cate, Jamie Hd

    2017-12-01

    Production of biofuels from lignocellulosic biomass remains an unsolved challenge in industrial biotechnology. Efforts to use yeast for conversion face the question of which host organism to use, counterbalancing the ease of genetic manipulation with the promise of robust industrial phenotypes. Saccharomyces cerevisiae remains the premier host for metabolic engineering of biofuel pathways, due to its many genetic, systems and synthetic biology tools. Numerous engineering strategies for expanding substrate ranges and diversifying products of S. cerevisiae have been developed. Other yeasts generally lack these tools, yet harbor superior phenotypes that could be exploited in the harsh processes required for lignocellulosic biofuel production. These include thermotolerance, resistance to toxic compounds generated during plant biomass deconstruction, and wider carbon consumption capabilities. Although promising, these yeasts have yet to be widely exploited. By contrast, oleaginous yeasts such as Yarrowia lipolytica capable of producing high titers of lipids are rapidly advancing in terms of the tools available for their metabolic manipulation. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Oxidative metabolism of 5-o-caffeoylquinic acid (chlorogenic acid), a bioactive natural product, by metalloporphyrin and rat liver mitochondria.

    dos Santos, Michel D; Martins, Patrícia R; dos Santos, Pierre A; Bortocan, Renato; Iamamoto, Y; Lopes, Norberto P

    2005-09-01

    Synthetic metalloporphyrins, in the presence of monooxygen donors, are known to mimic the various reactions of cytochrome P450 enzymes systems in the oxidation and oxygenation of various drugs and biologically active compounds. This paper reports an HPLC-MS-MS investigation of chlorogenic acid (CGA) oxidation by iodosylbenzene using iron(III) tetraphenylporphyrin chloride as catalyst. The oxidation products have been detected by sequential MS analyses. In addition, CGA was submitted to an in vitro metabolism assay employing isolated rat liver mitochondria. The single oxidized product obtained from mitochondrial metabolism corresponds to the major product formed by the metalloporphyrin-catalyzed reaction. These results indicate that biomimetic oxidation reactions, in addition to in vitro metabolism assays employing isolated organs/organelles, could replace some in vivo metabolism studies, thus minimizing the problems related to the use of a large number of living animals in experimental research.

  17. Metabolic modelling of polyhydroxyalkanoate copolymers production by mixed microbial cultures

    Reis Maria AM

    2008-07-01

    Full Text Available Abstract Background This paper presents a metabolic model describing the production of polyhydroxyalkanoate (PHA copolymers in mixed microbial cultures, using mixtures of acetic and propionic acid as carbon source material. Material and energetic balances were established on the basis of previously elucidated metabolic pathways. Equations were derived for the theoretical yields for cell growth and PHA production on mixtures of acetic and propionic acid as functions of the oxidative phosphorylation efficiency, P/O ratio. The oxidative phosphorylation efficiency was estimated from rate measurements, which in turn allowed the estimation of the theoretical yield coefficients. Results The model was validated with experimental data collected in a sequencing batch reactor (SBR operated under varying feeding conditions: feeding of acetic and propionic acid separately (control experiments, and the feeding of acetic and propionic acid simultaneously. Two different feast and famine culture enrichment strategies were studied: (i either with acetate or (ii with propionate as carbon source material. Metabolic flux analysis (MFA was performed for the different feeding conditions and culture enrichment strategies. Flux balance analysis (FBA was used to calculate optimal feeding scenarios for high quality PHA polymers production, where it was found that a suitable polymer would be obtained when acetate is fed in excess and the feeding rate of propionate is limited to ~0.17 C-mol/(C-mol.h. The results were compared with published pure culture metabolic studies. Conclusion Acetate was more conducive toward the enrichment of a microbial culture with higher PHA storage fluxes and yields as compared to propionate. The P/O ratio was not only influenced by the selected microbial culture, but also by the carbon substrate fed to each culture, where higher P/O ratio values were consistently observed for acetate than propionate. MFA studies suggest that when mixtures of

  18. Changes in pyridine metabolism profile during growth of trigonelline-forming Lotus japonicus cell cultures.

    Yin, Yuling; Matsui, Ayu; Sakuta, Masaaki; Ashihara, Hiroshi

    2008-12-01

    Changes in the profile of pyridine metabolism during growth of cells were investigated using trigonelline-forming suspension-cultured cells of Lotus japonicus. Activity of the de novo and salvage pathways of NAD biosynthesis was estimated from the in situ metabolism of [(3)H] quinolinic acid and [(14)C] nicotinamide. Maximum activity of the de novo pathway for NAD synthesis was found in the exponential growth phase, whereas activity of the salvage pathway was increased in the lag phase of cell growth. Expression profiles of some genes related to pyridine metabolism were examined using the expression sequence tags obtained from the L. japonicus database. Transcript levels of NaPRT and NIC, encoding salvage enzymes, were enhanced in the lag phase of cell growth, whereas the maximum expression of NADS was found in the exponential growth phase. Correspondingly, the activities of the salvage enzymes, nicotinate phosphoribosyltransferase (EC 2.4.2.11) and nicotinamidase (EC 3.5.1.19), increased one day after transfer of the stationary phase cells to the fresh medium. The greatest in situ trigonelline synthesis, both from [(3)H] quinolinic acid and [(14)C] nicotinamide, was found in the stationary phase of cell growth. The role of trigonelline in leguminous plants is discussed.

  19. Efficient protein production by yeast requires global tuning of metabolism

    Huang, Mingtao; Bao, Jichen; Hallstrom, Bjorn M.

    2017-01-01

    The biotech industry relies on cell factories for production of pharmaceutical proteins, of which several are among the top-selling medicines. There is, therefore, considerable interest in improving the efficiency of protein production by cell factories. Protein secretion involves numerous...... intracellular processes with many underlying mechanisms still remaining unclear. Here, we use RNA-seq to study the genome-wide transcriptional response to protein secretion in mutant yeast strains. We find that many cellular processes have to be attuned to support efficient protein secretion. In particular...... that by tuning metabolism cells are able to efficiently secrete recombinant proteins. Our findings provide increased understanding of which cellular regulations and pathways are associated with efficient protein secretion....

  20. Alteration of Fatty-Acid-Metabolizing Enzymes Affects Mitochondrial Form and Function in Hereditary Spastic Paraplegia

    Tesson, Christelle; Nawara, Magdalena; Salih, Mustafa A.M.; Rossignol, Rodrigue; Zaki, Maha S.; Al Balwi, Mohammed; Schule, Rebecca; Mignot, Cyril; Obre, Emilie; Bouhouche, Ahmed; Santorelli, Filippo M.; Durand, Christelle M.; Oteyza, Andrés Caballero; El-Hachimi, Khalid H.; Al Drees, Abdulmajeed; Bouslam, Naima; Lamari, Foudil; Elmalik, Salah A.; Kabiraj, Mohammad M.; Seidahmed, Mohammed Z.; Esteves, Typhaine; Gaussen, Marion; Monin, Marie-Lorraine; Gyapay, Gabor; Lechner, Doris; Gonzalez, Michael; Depienne, Christel; Mochel, Fanny; Lavie, Julie; Schols, Ludger; Lacombe, Didier; Yahyaoui, Mohamed; Al Abdulkareem, Ibrahim; Zuchner, Stephan; Yamashita, Atsushi; Benomar, Ali; Goizet, Cyril; Durr, Alexandra; Gleeson, Joseph G.; Darios, Frederic; Brice, Alexis; Stevanin, Giovanni

    2012-01-01

    Hereditary spastic paraplegia (HSP) is considered one of the most heterogeneous groups of neurological disorders, both clinically and genetically. The disease comprises pure and complex forms that clinically include slowly progressive lower-limb spasticity resulting from degeneration of the corticospinal tract. At least 48 loci accounting for these diseases have been mapped to date, and mutations have been identified in 22 genes, most of which play a role in intracellular trafficking. Here, we identified mutations in two functionally related genes (DDHD1 and CYP2U1) in individuals with autosomal-recessive forms of HSP by using either the classical positional cloning or a combination of whole-genome linkage mapping and next-generation sequencing. Interestingly, three subjects with CYP2U1 mutations presented with a thin corpus callosum, white-matter abnormalities, and/or calcification of the basal ganglia. These genes code for two enzymes involved in fatty-acid metabolism, and we have demonstrated in human cells that the HSP pathophysiology includes alteration of mitochondrial architecture and bioenergetics with increased oxidative stress. Our combined results focus attention on lipid metabolism as a critical HSP pathway with a deleterious impact on mitochondrial bioenergetic function. PMID:23176821

  1. Metabolic engineering of carbon overflow metabolism of Bacillus subtilis for improved N-acetyl-glucosamine production.

    Ma, Wenlong; Liu, Yanfeng; Shin, Hyun-Dong; Li, Jianghua; Chen, Jian; Du, Guocheng; Liu, Long

    2018-02-01

    Bacillus subtilis is widely used as cell factories for the production of important industrial biochemicals. Although many studies have demonstrated the effects of organic acidic byproducts, such as acetate, on microbial fermentation, little is known about the effects of blocking the neutral byproduct overflow, such as acetoin, on bioproduction. In this study, we focused on the influences of modulating overflow metabolism on the production of N-acetyl-d-glucosamine (GlcNAc) in engineered B. subtilis. We found that acetoin overflow competes with GlcNAc production, and blocking acetoin overflow increased GlcNAc titer and yield by 1.38- and 1.39-fold, reaching 48.9 g/L and 0.32 g GlcNAc/g glucose, respectively. Further blocking acetate overflow inhibited cell growth and GlcNAc production may be induced by inhibiting glucose uptake. Taken together, our results show that blocking acetoin overflow is a promising strategy for enhancing GlcNAc production. The strategies developed in this work may be useful for engineering strains of B. subtilis for producing other important biochemicals. Copyright © 2017. Published by Elsevier Ltd.

  2. Metabolic engineering of Escherichia coli for production of mixed-acid fermentation end products

    Andreas Hartmut Förster

    2014-05-01

    Full Text Available Mixed-acid fermentation end products have numerous applications in biotechnology. This is probably the main driving force for the development of multiple strains that are supposed to produce individual end products with high yields. The process of engineering Escherichia coli strains for applied production of ethanol, lactate, succinate, or acetate was initiated several decades ago and is still ongoing. This review follows the path of strain development from the general characteristics of aerobic versus anaerobic metabolism over the regulatory machinery that enables the different metabolic routes. Thereafter, major improvements for broadening the substrate spectrum of Escherichia coli towards cheap carbon sources like molasses or lignocellulose are highlighted before major routes of strain development for the production of ethanol, acetate, lactate and succinate are presented.

  3. Production of press moulds by plasma spray forming process

    Borisov, Y.; Myakota, I.; Polyakov, S.

    2001-01-01

    Plasma spray forming process for production of press moulds which are used for manufacture of articles from plastics was developed. The press moulds were produced by plasma spraying of Cu-Al-Fe-alloy powder on surface of a master model. The master models were made from non-metallic materials with heat resistance below 70 C (wood, gypsum etc). Double cooling system which provides for a control of surface model temperature and quenching conditions of sprayed material was designed. It made possible on the one hand to support model surface temperature below 70 C and on the other hand to provide for temperature conditions of martensite transformation in Cu-Al-system with a fixation of metastable ductile α + β 1 -phase. This allowed to decrease residual stresses in sprayed layer (up to 0,5-2,5 MPa), to increase microhardness of the coating material (up to 1200-1800 MPa) and its ductility (σ B = 70-105 MPa, δ = 6-12 %). This plasma spray forming process makes possible to spray thick layers (5-20 mm and more) without their cracking and deformation. The process is used for a production of press moulds which are applied in shoes industry, for fabrication of toys, souvenirs etc. (author)

  4. Metabolic Engineering of Oleaginous Yeasts for Fatty Alcohol Production

    Wang, Wei; Wei, Hui; Knoshaug, Eric; Van Wychen, Stefanie; Xu, Qi; Himmel, Michael E.; Zhang, Min

    2016-04-25

    To develop pathways for advanced biological upgrading of sugars to hydrocarbons, we are seeking biological approaches to produce high carbon efficiency intermediates amenable to separations and catalytic upgrading to hydrocarbon fuels. In this study, we successfully demonstrated fatty alcohol production by oleaginous yeasts Yarrowia lipolytica and Lipomyces starkeyi by expressing a bacteria-derived fatty acyl-CoA reductase (FAR). Moreover, we find higher extracellular distribution of fatty alcohols produced by FAR-expressing L. starkeyi strain as compared to Y. lipolytica strain, which would benefit the downstream product recovery process. In both oleaginous yeasts, long chain length saturated fatty alcohols were predominant, accounting for more than 85% of the total fatty alcohols produced. To the best of our knowledge, this is the first report of fatty alcohol production in L. starkeyi. Taken together, our work demonstrates that in addition to Y. lipolytica, L. starkeyi can also serve as a platform organism for production of fatty acid-derived biofuels and bioproducts via metabolic engineering. We believe strain and process development both will significantly contribute to our goal of producing scalable and cost-effective fatty alcohols from renewable biomass.

  5. Metabolic engineering of Dunaliella salina for production of ketocarotenoids.

    Anila, N; Simon, Daris P; Chandrashekar, Arun; Ravishankar, G A; Sarada, R

    2016-03-01

    Dunaliella is a commercially important marine alga producing high amount of β-carotene. The use of Dunaliella as a potential transgenic system for the production of recombinant proteins has been recently recognized. The present study reports for the first time the metabolic engineering of carotenoid biosynthesis in Dunaliella salina for ketocarotenoid production. The pathway modification included the introduction of a bkt gene from H. pluvialis encoding β-carotene ketolase (4,4'β-oxygenase) along with chloroplast targeting for the production of ketocarotenoids. The bkt under the control of Dunaliella Rubisco smaller subunit promoter along with its transit peptide sequence was introduced into the alga through standardized Agrobacterium-mediated transformation procedure. The selected transformants were confirmed using GFP and GUS expression, PCR and southern blot analysis. A notable upregulation of the endogenous hydroxylase level of transformants was observed where the BKT expression was higher in nutrient-limiting conditions. Carotenoid analysis of the transformants through HPLC and MS analysis showed the presence of astaxanthin and canthaxanthin with maximum content of 3.5 and 1.9 µg/g DW, respectively. The present study reports the feasibility of using D. salina for the production of ketocarotenoids including astaxanthin.

  6. Metabolic production of amphetamine following administration of clobenzorex.

    Valtier, S; Cody, J T

    1999-01-01

    Many of the anorectic drugs that are metabolized to amphetamine and/or methamphetamine pose significant concerns in the interpretation of amphetamine-positive drug testing results. One of these drugs--clobenzorex--has been shown to produce amphetamine. Thirty milligrams of clobenzorex hydrochloride, in the form of a single Asenlix capsule (Roussel, Mexico), were administered orally to five human volunteers with no history of amphetamine, methamphetamine or clobenzorex use. Following administration, urine samples (total void volume) were collected ad lib for seven days and pH, specific gravity and creatinine values were determined. To determine the excretion profile of amphetamine and parent drug, samples were extracted, derivatized, and analyzed by gas chromatography/mass spectrometry (GC/MS) using a standard amphetamine procedure with additional monitoring of ions at m/z 91, 118, 125 and 364 for the detection of clobenzorex. Peak concentrations of amphetamine were detected at 4 to 19 h postdose and ranged from approximately 715 to 2474 ng/mL amphetamine. Amphetamine could be detected (> 5 ng/mL) in the urine in one subject for up to 116 h postdose. GC/MS was also used to determine the enantiomeric composition of the metabolite, amphetamine. This analysis revealed the metabolically derived amphetamine was only the d-enantiomer. This differs from previous literature which indicates clobenzorex is the racemic N-orthochlorobenzyl derivative of amphetamine.

  7. 48 CFR 53.301-1427 - Standard Form 1427, Inventory Schedule A-Construction Sheet (Metals in Mill Product Form).

    2010-10-01

    ... 48 Federal Acquisition Regulations System 2 2010-10-01 2010-10-01 false Standard Form 1427, Inventory Schedule A-Construction Sheet (Metals in Mill Product Form). 53.301-1427 Section 53.301-1427... Illustrations of Forms 53.301-1427 Standard Form 1427, Inventory Schedule A—Construction Sheet (Metals in Mill...

  8. Nuclear reactor structural material forming less radioactive corrosion product

    Nakazawa, Hiroshi.

    1988-01-01

    Purpose: To provide nuclear reactor structural materials forming less radioactive corrosion products. Constitution: Ni-based alloys such as inconel alloy 718, 600 or inconel alloy 750 and 690 having excellent corrosion resistance and mechanical property even in coolants at high temperature and high pressure have generally been used as nuclear reactor structural materials. However, even such materials yield corrosion products being attacked by coolants circulating in the nuclear reactor, which produce by neutron irradiation radioactive corrosion products, that are deposited in primary circuit pipeways to constitute exposure sources. The present invention dissolves dissolves this problems by providing less activating nuclear reactor structural materials. That is, taking notice on the fact that Ni-58 contained generally by 68 % in Ni changes into Co-58 under irradiation of neutron thereby causing activation, the surface of nuclear reactor structural materials is applied with Ni plating by using Ni with a reduced content of Ni-58 isotopes. Accordingly, increase in the radiation level of the nuclear reactor structural materials can be inhibited. (K.M.)

  9. Metabolic engineering of Escherichia coli for the production of riboflavin

    2014-01-01

    Background Riboflavin (vitamin B2), the precursor of the flavin cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), is used commercially as an animal feed supplement and food colorant. E. coli is a robust host for various genetic manipulations and has been employed for efficient production of biofuels, polymers, amino acids, and bulk chemicals. Thus, the aim of this study was to understand the metabolic capacity of E. coli for the riboflavin production by modification of central metabolism, riboflavin biosynthesis pathway and optimization of the fermentation conditions. Results The basic producer RF01S, in which the riboflavin biosynthesis genes ribABDEC from E. coli were overexpressed under the control of the inducible trc promoter, could accumulate 229.1 mg/L of riboflavin. Further engineering was performed by examining the impact of expression of zwf (encodes glucose 6-phosphate dehydrogenase) and gnd (encodes 6-phosphogluconate dehydrogenase) from Corynebacterium glutamicum and pgl (encodes 6-phosphogluconolactonase) from E. coli on riboflavin production. Deleting pgi (encodes glucose-6-phosphate isomerase) and genes of Entner-Doudoroff (ED) pathway successfully redirected the carbon flux into the oxidative pentose phosphate pathway, and overexpressing the acs (encodes acetyl-CoA synthetase) reduced the acetate accumulation. These modifications increased riboflavin production to 585.2 mg/L. By further modulating the expression of ribF (encodes riboflavin kinase) for reducing the conversion of riboflavin to FMN in RF05S, the final engineering strain RF05S-M40 could produce 1036.1 mg/L riboflavin in LB medium at 37°C. After optimizing the fermentation conditions, strain RF05S-M40 produced 2702.8 mg/L riboflavin in the optimized semi-defined medium, which was a value nearly 12-fold higher than that of RF01S, with a yield of 137.5 mg riboflavin/g glucose. Conclusions The engineered strain RF05S-M40 has the highest yield among all

  10. Metabolic engineering of Escherichia coli for the production of riboflavin.

    Lin, Zhenquan; Xu, Zhibo; Li, Yifan; Wang, Zhiwen; Chen, Tao; Zhao, Xueming

    2014-07-16

    Riboflavin (vitamin B2), the precursor of the flavin cofactors flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD), is used commercially as an animal feed supplement and food colorant. E. coli is a robust host for various genetic manipulations and has been employed for efficient production of biofuels, polymers, amino acids, and bulk chemicals. Thus, the aim of this study was to understand the metabolic capacity of E. coli for the riboflavin production by modification of central metabolism, riboflavin biosynthesis pathway and optimization of the fermentation conditions. The basic producer RF01S, in which the riboflavin biosynthesis genes ribABDEC from E. coli were overexpressed under the control of the inducible trc promoter, could accumulate 229.1 mg/L of riboflavin. Further engineering was performed by examining the impact of expression of zwf (encodes glucose 6-phosphate dehydrogenase) and gnd (encodes 6-phosphogluconate dehydrogenase) from Corynebacterium glutamicum and pgl (encodes 6-phosphogluconolactonase) from E. coli on riboflavin production. Deleting pgi (encodes glucose-6-phosphate isomerase) and genes of Entner-Doudoroff (ED) pathway successfully redirected the carbon flux into the oxidative pentose phosphate pathway, and overexpressing the acs (encodes acetyl-CoA synthetase) reduced the acetate accumulation. These modifications increased riboflavin production to 585.2 mg/L. By further modulating the expression of ribF (encodes riboflavin kinase) for reducing the conversion of riboflavin to FMN in RF05S, the final engineering strain RF05S-M40 could produce 1036.1 mg/L riboflavin in LB medium at 37°C. After optimizing the fermentation conditions, strain RF05S-M40 produced 2702.8 mg/L riboflavin in the optimized semi-defined medium, which was a value nearly 12-fold higher than that of RF01S, with a yield of 137.5 mg riboflavin/g glucose. The engineered strain RF05S-M40 has the highest yield among all reported riboflavin production

  11. Estimating product-to-product variations in metal forming using force measurements

    Havinga, Jos; van den Boogaard, Ton

    2017-10-01

    The limits of production accuracy of metal forming processes can be stretched by the development of control systems for compensation of product-to-product variations. Such systems require the use of measurements from each semi-finished product. These measurements must be used to estimate the final quality of each product. We propose to predict part of the product-to-product variations in multi-stage forming processes based on force measurements from previous process stages. The reasoning is that final product properties as well as process forces are expected to be correlated since they are both affected by material and process variation. In this study, an approach to construct a moving window process model based on historical data from the process is presented. These regression models can be built and updated in real-time during production. The approach is tested with data from a demonstrator process with cutting, deep drawing and bending stages. It is shown that part of the product-to-product variations in the process can be predicted with the developed process model.

  12. Acetone production with metabolically engineered strains of Acetobacterium woodii.

    Hoffmeister, Sabrina; Gerdom, Marzena; Bengelsdorf, Frank R; Linder, Sonja; Flüchter, Sebastian; Öztürk, Hatice; Blümke, Wilfried; May, Antje; Fischer, Ralf-Jörg; Bahl, Hubert; Dürre, Peter

    2016-07-01

    Expected depletion of oil and fossil resources urges the development of new alternative routes for the production of bulk chemicals and fuels beyond petroleum resources. In this study, the clostridial acetone pathway was used for the formation of acetone in the acetogenic bacterium Acetobacterium woodii. The acetone production operon (APO) containing the genes thlA (encoding thiolase A), ctfA/ctfB (encoding CoA transferase), and adc (encoding acetoacetate decarboxylase) from Clostridium acetobutylicum were cloned under the control of the thlA promoter into four vectors having different replicons for Gram-positives (pIP404, pBP1, pCB102, and pCD6). Stable replication was observed for all constructs. A. woodii [pJIR_actthlA] achieved the maximal acetone concentration under autotrophic conditions (15.2±3.4mM). Promoter sequences of the genes ackA from A. woodii and pta-ack from C. ljungdahlii were determined by primer extension (PEX) and cloned upstream of the APO. The highest acetone production in recombinant A. woodii cells was achieved using the promoters PthlA and Ppta-ack. Batch fermentations using A. woodii [pMTL84151_actthlA] in a bioreactor revealed that acetate concentration had an effect on the acetone production, due to the high Km value of the CoA transferase. In order to establish consistent acetate concentration within the bioreactor and to increase biomass, a continuous fermentation process for A. woodii was developed. Thus, acetone productivity of the strain A. woodii [pMTL84151_actthlA] was increased from 1.2mgL(-1)h(-1) in bottle fermentation to 26.4mgL(-1)h(-1) in continuous gas fermentation. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  13. Immobilization of fission products in phosphate ceramic waste forms

    Singh, D.; Wagh, A. [Argonne National Lab., IL (United States)

    1997-10-01

    Chemically bonded phosphate ceramics (CBPCs) have several advantages that make them ideal candidates for containing radioactive and hazardous wastes. In general, phosphates have high solid-solution capacities for incorporating radionuclides, as evidenced by several phosphates (e.g., monazites and apatites) that are natural analogs of radioactive and rare-earth elements. The phosphates have high radiation stability, are refractory, and will not degrade in the presence of internal heating by fission products. Dense and hard CBPCs can be fabricated inexpensively and at low temperature by acid-base reactions between an inorganic oxide/hydroxide powder and either phosphoric acid or an acid-phosphate solution. The resulting phosphates are extremely insoluble in aqueous media and have excellent long-term durability. CBPCs offer the dual stabilization mechanisms of chemical fixation and physical encapsulation, resulting in superior waste forms. The goal of this task is develop and demonstrate the feasibility of CBPCs for S/S of wastes containing fission products. The focus of this work is to develop a low-temperature CBPC immobilization system for eluted {sup 99}Tc wastes from sorption processes.

  14. Adaptations in the glucose metabolism of procyclic Trypanosoma brucei isolates from Tsetse flies and during differentiation of bloodstream forms.

    van Grinsven, K.W.A.; van den Abbeele, J.; van den Bossche, P.; van Hellemond, J.J.; Tielens, A.G.M.

    2009-01-01

    Procyclic forms of Trypanosoma brucei isolated from the midguts of infected tsetse flies, or freshly transformed from a strain that is close to field isolates, do not use a complete Krebs cycle. Furthermore, short stumpy bloodstream forms produce acetate and are apparently metabolically preadapted

  15. High-level waste-form-product performance evaluation

    Bernadzikowski, T.A.; Allender, J.S.; Stone, J.A.; Gordon, D.E.; Gould, T.H. Jr.; Westberry, C.F. III.

    1982-01-01

    Seven candidate waste forms were evaluated for immobilization and geologic disposal of high-level radioactive wastes. The waste forms were compared on the basis of leach resistance, mechanical stability, and waste loading. All forms performed well at leaching temperatures of 40, 90, and 150 0 C. Ceramic forms ranked highest, followed by glasses, a metal matrix form, and concrete. 11 tables

  16. Metabolic Engineering and Modeling of Metabolic Pathways to Improve Hydrogen Production by Photosynthetic Bacteria

    Jiao, Y. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Navid, A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2014-12-19

    traits act as the biocatalysts of the process designed to both enhance the system efficiency of CO2 fixation and the net hydrogen production rate. Additionally we applied metabolic engineering approaches guided by computational modeling for the chosen model microorganisms to enable efficient hydrogen production.

  17. Metabolic engineering of ethanol production in Thermoanaerobacter mathranii

    Shou Yao

    2010-11-15

    Strain BG1 is a xylanolytic, thermophilic, anaerobic, Gram-positive bacterium originally isolated from an Icelandic hot spring. The strain belongs to the species Thermoanaerobacter mathranii. The strain ferments glucose, xylose, arabinose, galactose and mannose simultaneously and produces ethanol, acetate, lactate, CO{sub 2}, and H2 as fermentation end-products. As a potential ethanol producer from lignocellulosic biomass, tailor-made BG1 strain with the metabolism redirected to produce ethanol is needed. Metabolic engineering of T. mathranii BG1 is therefore necessary to improve ethanol production. Strain BG1 contains four alcohol dehydrogenase (ADH) encoding genes. They are adhA, adhB, bdhA and adhE encoding primary alcohol dehydrogenase, secondary alcohol dehydrogenase, butanol dehydrogenase and bifunctional alcohol/acetaldehyde dehydrogenase, respectively. The presence in an organism of multiple alcohol dehydrogenases with overlapping specificities makes the determination of the specific role of each ADH difficult. Deletion of each individual adh gene in the strain revealed that the adhE deficient mutant strain fails to produce ethanol as the fermentation product. The bifunctional alcohol/acetaldehyde dehydrogenase, AdhE, is therefore proposed responsible for ethanol production in T. mathranii BG1, by catalyzing sequential NADH-dependent reductions of acetyl-CoA to acetaldehyde and then to ethanol under fermentative conditions. Moreover, AdhE was conditionally expressed from a xylose-induced promoter in a recombinant strain (BG1E1) with a concomitant deletion of a lactate dehydrogenase. Over-expression of AdhE in strain BG1E1 with xylose as a substrate facilitates the production of ethanol at an increased yield. With a cofactor-dependent ethanol production pathway in T. mathranii BG1, it may become crucial to regenerate cofactor to increase the ethanol yield. Feeding the cells with a more reduced carbon source, such as mannitol, was shown to increase ethanol

  18. Transcriptomic Changes in Response to Putrescine Production in Metabolically Engineered Corynebacterium glutamicum

    Zhen Li

    2017-10-01

    Full Text Available Putrescine is widely used in industrial production of bioplastics, pharmaceuticals, agrochemicals, and surfactants. Although engineered Corynebacterium glutamicum has been successfully used to produce high levels of putrescine, the overall cellular physiological and metabolic changes caused by overproduction of putrescine remains unclear. To reveal the transcriptional changes that occur in response to putrescine production in an engineered C. glutamicum strain, a comparative transcriptomic analysis was carried out. Overproduction of putrescine resulted in transcriptional downregulation of genes involved in glycolysis; the TCA cycle, pyruvate degradation, biosynthesis of some amino acids, oxidative phosphorylation; vitamin biosynthesis (thiamine and vitamin 6, metabolism of purine, pyrimidine and sulfur, and ATP-, NAD-, and NADPH-consuming enzymes. The transcriptional levels of genes involved in ornithine biosynthesis and NADPH-forming related enzymes were significantly upregulated in the putrescine producing C. glutamicum strain PUT-ALE. Comparative transcriptomic analysis provided some genetic modification strategies to further improve putrescine production. Repressing ATP- and NADPH-consuming enzyme coding gene expression via CRISPRi enhanced putrescine production.

  19. Transcriptomic Changes in Response to Putrescine Production in Metabolically Engineered Corynebacterium glutamicum

    Li, Zhen; Liu, Jian-Zhong

    2017-01-01

    Putrescine is widely used in industrial production of bioplastics, pharmaceuticals, agrochemicals, and surfactants. Although engineered Corynebacterium glutamicum has been successfully used to produce high levels of putrescine, the overall cellular physiological and metabolic changes caused by overproduction of putrescine remains unclear. To reveal the transcriptional changes that occur in response to putrescine production in an engineered C. glutamicum strain, a comparative transcriptomic analysis was carried out. Overproduction of putrescine resulted in transcriptional downregulation of genes involved in glycolysis; the TCA cycle, pyruvate degradation, biosynthesis of some amino acids, oxidative phosphorylation; vitamin biosynthesis (thiamine and vitamin 6), metabolism of purine, pyrimidine and sulfur, and ATP-, NAD-, and NADPH-consuming enzymes. The transcriptional levels of genes involved in ornithine biosynthesis and NADPH-forming related enzymes were significantly upregulated in the putrescine producing C. glutamicum strain PUT-ALE. Comparative transcriptomic analysis provided some genetic modification strategies to further improve putrescine production. Repressing ATP- and NADPH-consuming enzyme coding gene expression via CRISPRi enhanced putrescine production. PMID:29089930

  20. Considerations on pig models for appetite, metabolic syndrome and obese type 2 diabetes: Form food intake to metabolic disease

    Koopmans, S.J.; Schuurman, T.

    2015-01-01

    (Mini)pigs have proven to be a valuable animal model in nutritional, metabolic and cardiovascular research and in some other biomedical research areas (toxicology, neurobiology). The large resemblance of (neuro)anatomy, the gastro-intestinal tract, body size, body composition, and the omnivorous

  1. Kinetic modeling of cell metabolism for microbial production.

    Costa, Rafael S; Hartmann, Andras; Vinga, Susana

    2016-02-10

    Kinetic models of cellular metabolism are important tools for the rational design of metabolic engineering strategies and to explain properties of complex biological systems. The recent developments in high-throughput experimental data are leading to new computational approaches for building kinetic models of metabolism. Herein, we briefly survey the available databases, standards and software tools that can be applied for kinetic models of metabolism. In addition, we give an overview about recently developed ordinary differential equations (ODE)-based kinetic models of metabolism and some of the main applications of such models are illustrated in guiding metabolic engineering design. Finally, we review the kinetic modeling approaches of large-scale networks that are emerging, discussing their main advantages, challenges and limitations. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Precursors and metabolic pathway for guaiacol production by Alicyclobacillus acidoterrestris.

    Cai, Rui; Yuan, Yahong; Wang, Zhouli; Guo, Chunfeng; Liu, Bin; Liu, Laping; Wang, Yutang; Yue, Tianli

    2015-12-02

    Alicyclobacillus acidoterrestris has recently received much attention due to its implication in the spoilage of pasteurized fruit juices, which was manifested by the production of guaiacol. Vanillic acid and vanillin have been accepted as the biochemical precursors of guaiacol in fruit juices. The purpose of this study was to try to find other precursors and elucidate details about the conversion of vanillic acid and vanillin to guaiacol by A. acidoterrestris. Four potential substrates including ferulic acid, catechol, phenylalanine and tyrosine were analyzed, but they could not be metabolized to guaiacol by all the thirty A. acidoterrestris strains tested. Resting cell studies and enzyme assays demonstrated that vanillin was reduced to vanillyl alcohol by NADPH-dependent vanillin reductase and oxidized to vanillic acid by NAD(P)(+)-dependent vanillin dehydrogenases in A. acidoterrestris DSM 3923. Vanillic acid underwent a nonoxidative decarboxylation to guaiacol. The reversible vanillic acid decarboxylase involved was oxygen insensitive and pyridine nucleotide-independent. Copyright © 2015. Published by Elsevier B.V.

  3. Metabolic flexibility revealed in the genome of the cyst-forming α-1 proteobacterium Rhodospirillum centenum

    Matthies Heather J

    2010-05-01

    Full Text Available Abstract Background Rhodospirillum centenum is a photosynthetic non-sulfur purple bacterium that favors growth in an anoxygenic, photosynthetic N2-fixing environment. It is emerging as a genetically amenable model organism for molecular genetic analysis of cyst formation, photosynthesis, phototaxis, and cellular development. Here, we present an analysis of the genome of this bacterium. Results R. centenum contains a singular circular chromosome of 4,355,548 base pairs in size harboring 4,105 genes. It has an intact Calvin cycle with two forms of Rubisco, as well as a gene encoding phosphoenolpyruvate carboxylase (PEPC for mixotrophic CO2 fixation. This dual carbon-fixation system may be required for regulating internal carbon flux to facilitate bacterial nitrogen assimilation. Enzymatic reactions associated with arsenate and mercuric detoxification are rare or unique compared to other purple bacteria. Among numerous newly identified signal transduction proteins, of particular interest is a putative bacteriophytochrome that is phylogenetically distinct from a previously characterized R. centenum phytochrome, Ppr. Genes encoding proteins involved in chemotaxis as well as a sophisticated dual flagellar system have also been mapped. Conclusions Remarkable metabolic versatility and a superior capability for photoautotrophic carbon assimilation is evident in R. centenum.

  4. The oxidized form of vitamin C, dehydroascorbic acid, regulates neuronal energy metabolism.

    Cisternas, Pedro; Silva-Alvarez, Carmen; Martínez, Fernando; Fernandez, Emilio; Ferrada, Luciano; Oyarce, Karina; Salazar, Katterine; Bolaños, Juan P; Nualart, Francisco

    2014-05-01

    Vitamin C is an essential factor for neuronal function and survival, existing in two redox states, ascorbic acid (AA), and its oxidized form, dehydroascorbic acid (DHA). Here, we show uptake of both AA and DHA by primary cultures of rat brain cortical neurons. Moreover, we show that most intracellular AA was rapidly oxidized to DHA. Intracellular DHA induced a rapid and dramatic decrease in reduced glutathione that was immediately followed by a spontaneous recovery. This transient decrease in glutathione oxidation was preceded by an increase in the rate of glucose oxidation through the pentose phosphate pathway (PPP), and a concomitant decrease in glucose oxidation through glycolysis. DHA stimulated the activity of glucose-6-phosphate dehydrogenase, the rate-limiting enzyme of the PPP. Furthermore, we found that DHA stimulated the rate of lactate uptake by neurons in a time- and dose-dependent manner. Thus, DHA is a novel modulator of neuronal energy metabolism by facilitating the utilization of glucose through the PPP for antioxidant purposes. © 2014 International Society for Neurochemistry.

  5. A state of the art of metabolic networks of unicellular microalgae and cyanobacteria for biofuel production.

    Baroukh, Caroline; Muñoz-Tamayo, Rafael; Steyer, Jean-Philippe; Bernard, Olivier

    2015-07-01

    The most promising and yet challenging application of microalgae and cyanobacteria is the production of renewable energy: biodiesel from microalgae triacylglycerols and bioethanol from cyanobacteria carbohydrates. A thorough understanding of microalgal and cyanobacterial metabolism is necessary to master and optimize biofuel production yields. To this end, systems biology and metabolic modeling have proven to be very efficient tools if supported by an accurate knowledge of the metabolic network. However, unlike heterotrophic microorganisms that utilize the same substrate for energy and as carbon source, microalgae and cyanobacteria require light for energy and inorganic carbon (CO2 or bicarbonate) as carbon source. This double specificity, together with the complex mechanisms of light capture, makes the representation of metabolic network nonstandard. Here, we review the existing metabolic networks of photoautotrophic microalgae and cyanobacteria. We highlight how these networks have been useful for gaining insight on photoautotrophic metabolism. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  6. Effects of metabolic modifiers such as carnitines, coenzyme Q10, and PUFAs against different forms of neurotoxic insults: metabolic inhibitors, MPTP, and methamphetamine.

    Virmani, Ashraf; Gaetani, Franco; Binienda, Zbigniew

    2005-08-01

    , L-C, together with its acetyl ester, acetyl-L-carnitine (ALC), also participates in the control of the mitochondrial acyl-CoA/CoA ratio, peroxisomal oxidation of fatty acids, and production of ketone bodies. A deficiency of carnitine is known to have major deleterious effects on the CNS. We have examined L-C and its acetylated derivative, ALC, as potential neuroprotective compounds using various known metabolic inhibitors, as well as against drugs of abuse such as methamphetamine.

  7. Oxidation kinetics of reaction products formed in uranium metal corrosion

    Totemeier, T. C.

    1998-01-01

    The oxidation behavior of uranium metal ZPPR fuel corrosion products in environments of Ar-4%O 2 and Ar-20%O 2 were studied using thermo-gravimetric analysis (TGA). These tests were performed to extend earlier work in this area specifically, to assess plate-to-plate variations in corrosion product properties and the effect of oxygen concentration on oxidation behavior. The corrosion products from two relatively severely corroded plates were similar, while the products from a relatively intact plate were not reactive. Oxygen concentration strongly affected the burning rate of reactive products, but had little effect on low-temperature oxidation rates

  8. Oxidation kinetics of reaction products formed in uranium metal corrosion.

    Totemeier, T. C.

    1998-04-22

    The oxidation behavior of uranium metal ZPPR fuel corrosion products in environments of Ar-4%O{sub 2} and Ar-20%O{sub 2} were studied using thermo-gravimetric analysis (TGA). These tests were performed to extend earlier work in this area specifically, to assess plate-to-plate variations in corrosion product properties and the effect of oxygen concentration on oxidation behavior. The corrosion products from two relatively severely corroded plates were similar, while the products from a relatively intact plate were not reactive. Oxygen concentration strongly affected the burning rate of reactive products, but had little effect on low-temperature oxidation rates.

  9. A modular metabolic engineering approach for the production of 1,2-propanediol from glycerol by Saccharomyces cerevisiae

    Islam, Zia-Ul; Klein, Mathias; Aßkamp, Maximilian R.

    2017-01-01

    Compared to sugars, a major advantage of using glycerol as a feedstock for industrial bioprocesses is the fact that this molecule is more reduced than sugars. A compound whose biotechnological production might greatly profit from the substrate's higher reducing power is 1,2-propanediol (1,2-PDO...... as the nitrogen source. When nutrient-limited complex YG medium was used, significant amounts of 1,2-PDO were formed and it became obvious that the concerted supply of NADH and DHAP are essential for boosting 1,2-PDO production. Additionally, optimizing the flux into the MG pathway improved 1,2-PDO formation...... on both the strength of the metabolic switch towards fermentation in general (as indicated by the production of ethanol and 1,2-PDO) and on the ratio at which these two fermentation products were formed. For example, virtually no 1,2-PDO but only ethanol was formed in synthetic glycerol medium with urea...

  10. The role of metabolic engineering in the production of secondary metabolites

    Nielsen, Jens Bredal

    1998-01-01

    In the production of secondary metabolites yield and productivity are the most important design parameters. The focus is therefore to direct the carbon fluxes towards the product of interest, and this can be obtained through metabolic engineering whereby directed genetic changes are introduced...... into the production strain. In this process it is, however, important to analyze the metabolic network through measurement of the intracellular metabolites and the flux distributions. Besides playing an important role in the optimization of existing processes, metabolic engineering also offers the possibility...

  11. Production, transport, and metabolism of ethanol in eastern cottonwood

    MacDonald, R.C.

    1991-01-01

    In plant tissues, the production of acetaldehyde and ethanol are usually thought to occur as a mechanism to allow tolerance of hypoxic conditions. Acetaldehyde and ethanol were found to be common in vascular cambium and the transpiration stream of trees. Ethanol concentrations in the vascular cambium of Populus deltoides were not changed by placing logs from nonflooded trees in a pure oxygen environment for as long as 96 h, but increased by almost 3 orders of magnitude when exposed to low external pO 2 s. Ethanol is present in the xylem sap of flooded and nonflooded trees. Because of the constitutive presence of alcohol dehydrogenase in the mature leaves of woody plants, it was hypothesized that the leaves and shoots of trees had the ability to metabolize ethanol supplied by the transpiration stream. 1-[ 14 C]ethanol was supplied to excised leaves and shoots of Populus deltoides Bartr. in short- and long-term experiments. Greater than 99% of the radiolabel was incorporated into plant tissue in short-term experiments, with more than 95% of the label remaining in plant tissue after 24 h. Very little label reached the leaf mesophyll cells of excised shoots, as revealed by autoradiography. Radiolabel appeared primarily in the water- and chloroform-soluble fractions in short-term experiments, while in long-term experiments, label was also incorporated into protein. When labelled ethanol was supplied to excised petioles in a 5 min pulse, 41% of the label was incorporated into organic acids. Some label was also incorporated into amino acids, protein, and the chloroform-soluble fraction, with very little appearing in neutral sugars, starch, or the insoluble pellet. Labelled organic acids were separated by HPLC, and were comprised of acetate, isocitrate, α-ketoglutarate, and succinate. There was no apparent incorporation of label into phosphorylated compounds

  12. Metabolic clearance and blood production rates of estradiol in hyperthyroidism.

    Ridgway, E C; Longcope, C; Maloof, F

    1975-09-01

    The metabolic clearance rate of 17beta-estradiol (MCR2), the plasma levels of 17beta-estradiol (E2)1, sex-steroid binding globulin (SSBG), luteinizing hormone (LH) and follicle-stimulating hormone (FSH) were measured in 10 hyperthyroid subjects (7 men and 3 women). The blood production rate of 17beta-estradiol (PB2) was calculated for all subjects. Nine of the 10 hyperthyroid subjects had a decreased MCR2 which returned towards normal in 5 of the 6 subjects restudied following therapy. In all 10 subjects the levels of SSBG were increased when they were hyperthyroid and returned toward normal with therapy. It is concluded that the decrease in MCR2 is largely due to the increased binding of 17beta-estradiol to SSBG. In 7 of the 10 hyperthyroid the plasma E2 concentrations were normal whereas 3 had slightly elevated levels. In 8 of the 10 hyperthyroid the PB2 was within the normal range. Only 2 hyperthyroid subjects had slightly elevated PB2. In the 6 subjects who were restudied after therapy, there was no consistent change in PB2 which remained in the normal range in all cases. It is concluded that the MCR2 is decreased in most subjects with hyperthyroidism in association with an increase of SSBG. Despite this change in MCR2 there is no significant change in PB2. The increase in SSBG levels in hyperthyroidism appears to be a direct effect of the elevation of thyroid hormone activity and is not mediated through estrogen.

  13. Metabolic engineering approaches for production of biochemicals in food and medicinal plants.

    Wilson, Sarah A; Roberts, Susan C

    2014-04-01

    Historically, plants are a vital source of nutrients and pharmaceuticals. Recent advances in metabolic engineering have made it possible to not only increase the concentration of desired compounds, but also introduce novel biosynthetic pathways to a variety of species, allowing for enhanced nutritional or commercial value. To improve metabolic engineering capabilities, new transformation techniques have been developed to allow for gene specific silencing strategies or stacking of multiple genes within the same region of the chromosome. The 'omics' era has provided a new resource for elucidation of uncharacterized biosynthetic pathways, enabling novel metabolic engineering approaches. These resources are now allowing for advanced metabolic engineering of plant production systems, as well as the synthesis of increasingly complex products in engineered microbial hosts. The status of current metabolic engineering efforts is highlighted for the in vitro production of paclitaxel and the in vivo production of β-carotene in Golden Rice and other food crops. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. Metabolism

    ... Are More Common in People With Type 1 Diabetes Metabolic Syndrome Your Child's Weight Healthy Eating Endocrine System Blood Test: Basic Metabolic Panel (BMP) Activity: Endocrine System Growth Disorders Diabetes Center Thyroid Disorders Your Endocrine System Movie: Endocrine ...

  15. Immobilization of fission products in phosphate ceramic waste forms

    Singh, D.

    1996-01-01

    The goal of this project is to develop and demonstrate the feasibility of a novel low-temperature solidification/stabilization (S/S) technology for immobilizing waste streams containing fission products such as cesium, strontium, and technetium in a chemically bonded phosphate ceramic. This technology can immobilize partitioned tank wastes and decontaminate waste streams containing volatile fission products

  16. Metabolic Engineering toward Sustainable Production of Nylon-6.

    Turk, Stefan C H J; Kloosterman, Wigard P; Ninaber, Dennis K; Kolen, Karin P A M; Knutova, Julia; Suir, Erwin; Schürmann, Martin; Raemakers-Franken, Petronella C; Müller, Monika; de Wildeman, Stefaan M A; Raamsdonk, Leonie M; van der Pol, Ruud; Wu, Liang; Temudo, Margarida F; van der Hoeven, Rob A M; Akeroyd, Michiel; van der Stoel, Roland E; Noorman, Henk J; Bovenberg, Roel A L; Trefzer, Axel C

    2016-01-15

    Nylon-6 is a bulk polymer used for many applications. It consists of the non-natural building block 6-aminocaproic acid, the linear form of caprolactam. Via a retro-synthetic approach, two synthetic pathways were identified for the fermentative production of 6-aminocaproic acid. Both pathways require yet unreported novel biocatalytic steps. We demonstrated proof of these bioconversions by in vitro enzyme assays with a set of selected candidate proteins expressed in Escherichia coli. One of the biosynthetic pathways starts with 2-oxoglutarate and contains bioconversions of the ketoacid elongation pathway known from methanogenic archaea. This pathway was selected for implementation in E. coli and yielded 6-aminocaproic acid at levels up to 160 mg/L in lab-scale batch fermentations. The total amount of 6-aminocaproic acid and related intermediates generated by this pathway exceeded 2 g/L in lab-scale fed-batch fermentations, indicating its potential for further optimization toward large-scale sustainable production of nylon-6.

  17. Form gene clustering method about pan-ethnic-group products based on emotional semantic

    Chen, Dengkai; Ding, Jingjing; Gao, Minzhuo; Ma, Danping; Liu, Donghui

    2016-09-01

    The use of pan-ethnic-group products form knowledge primarily depends on a designer's subjective experience without user participation. The majority of studies primarily focus on the detection of the perceptual demands of consumers from the target product category. A pan-ethnic-group products form gene clustering method based on emotional semantic is constructed. Consumers' perceptual images of the pan-ethnic-group products are obtained by means of product form gene extraction and coding and computer aided product form clustering technology. A case of form gene clustering about the typical pan-ethnic-group products is investigated which indicates that the method is feasible. This paper opens up a new direction for the future development of product form design which improves the agility of product design process in the era of Industry 4.0.

  18. Metabolic clearance and production rates of human growth hormone

    Taylor, Andrew L.; Finster, Joseph L.; Mintz, Daniel H.

    1969-01-01

    The metabolic clearance rate (MCR) of human growth hormone (HGH) was determined by the constant infusion to equilibrium technique utilizing HGH-125I. 22 control subjects had a MCR of 229 ±52 ml/min (mean ±SD). No difference was evident between sexes, or between various age groups. Patients with acromegaly demonstrated normal MCR's. Moreover, acute elevations of plasma growth hormone concentrations in normal subjects did not alter the MCR of HGH. The MCR was relatively constant from day to day and within the day when subjects were evaluated in the supine position. In contrast, the assumption of the upright position was associated with a mean 24% decrease in the MCR. These results were contrasted with the MCR of HGH observed in a small number of patients with altered thyroid function or diabetes mellitus. In six patients with hypothyroidism the MCR (131 ±36 ml/min) was significantly decreased (P < 0.001); whereas the MCR in eight patients with hyperthyroidism (240 ±57 ml/min) did not differ from control subjects. The MCR in eight patients with insulin-independent diabetes mellitus (IID) (185 ±41 ml/min) and in eight patients with insulin-dependent diabetes mellitus (IDD) (136 ±31 ml/min) were significantly different from control subjects (P = < 0.05 and P = < 0.001, respectively). These data were interpreted to indicate that the plasma HGH-removing mechanism(s) is not saturated at physiologic plasma HGH levels, that plasma HGH levels alone may not permit distinction between variations in pituitary release of the hormone and its rate of clearance from the plasma, and that the estimation of the MCR of HGH may help clarify the mechanism of abnormal plasma HGH responses to various stimuli. Production rates of HGH (PR) in control subjects (347 ±173 mμg/min) were contrasted with hyperthyroid patients (529 ±242 mμg/min, P < 0.05), hypothyroid patients (160 ±69 mμg/min, P < 0.02), IID (245 ±100 mμg/min, NS), and IDD (363 ±153 mμg/min, NS). Considerable

  19. Metabolism of leukotriene B4 to dihydro and dihydro-oxo products by porcine leukocytes

    Powell, W.S.; Gravelle, F.

    1989-01-01

    Porcine leukocytes contain a novel pathway for the metabolism of leukotriene B4 (LTB4) which results in reduction of the conjugated triene chromophore to a conjugated diene. These cells converted LTB4 to two major metabolites, both of which exhibited maximal absorbance at 230 nm in their UV spectra. These products were purified by high pressure liquid chromatography and identified as 10, 11-dihydro-LTB4 and 10,11-dihydro-12-oxo-LTB4 on the basis of the mass spectra of various derivatives. The position of the double bond of LTB4 which had been reduced was established by cleaving the remaining double bonds of 10, 11-dihydro-LTB4 with ozone followed by oxidation or reduction of the resulting ozonide and analysis of the products by mass spectrometry. Experiments with deuterium-labeled substrate indicated that LTB4 could be directly converted to 10, 11-dihydro-LTB4 without the prior oxidation of either of its hydroxyl groups, as is required for the formation of dihydro metabolites of prostaglandins. Incubation of porcine leukocytes with 10, 11-dihydro-LTB4 and 10, 11-dihydro-12-oxo-LTB4 indicated that these two products can be interconverted and are in equilibrium with one another. The dihydro-oxo metabolite can therefore be formed from 10, 11-dihydro-LTB4, although we have not ruled out the possibility that it is also produced via 12-oxo-LTB4, which could be a transitory intermediate. These results indicate that porcine leukocytes contain a novel reductase/dehydrogenase pathway distinct from the pathway responsible for the metabolism of prostaglandins. This pathway is also different from the pathway in human polymorphonuclear leukocytes which converts 6-trans-isomers of LTB4 to dihydro products, since the latter pathway involves 5-oxo intermediates and results in a shift in the positions of the remaining double bonds

  20. Spatial separation of photosynthesis and ethanol production by cell type-specific metabolic engineering of filamentous cyanobacteria.

    Ehira, Shigeki; Takeuchi, Takuto; Higo, Akiyoshi

    2018-02-01

    Cyanobacteria, which perform oxygenic photosynthesis, have drawn attention as hosts for the direct production of biofuels and commodity chemicals from CO 2 and H 2 O using light energy. Although cyanobacteria capable of producing diverse chemicals have been generated by metabolic engineering, anaerobic non-photosynthetic culture conditions are often necessary for their production. In this study, we conducted cell type-specific metabolic engineering of the filamentous cyanobacterium Anabaena sp. PCC 7120, which forms a terminally differentiated cell called a heterocyst with a semi-regular spacing of 10-15 cells. Because heterocysts are specialized cells for nitrogen fixation, the intracellular oxygen level of heterocysts is maintained very low even when adjacent cells perform oxygenic photosynthesis. Pyruvate decarboxylase of Zymomonas mobilis and alcohol dehydrogenase of Synechocystis sp. PCC 6803 were exclusively expressed in heterocysts. Ethanol production was concomitant with nitrogen fixation in genetically engineered Anabaena sp. PCC 7120. Engineering of carbon metabolism in heterocysts improved ethanol production, and strain ET14, with an extra copy of the invB gene expressed from a heterocyst-specific promoter, produced 130.9 mg L -1 of ethanol after 9 days. ET14 produced 1681.9 mg L -1 of ethanol by increasing the CO 2 supply. Ethanol production per heterocyst cell was approximately threefold higher than that per cell of unicellular cyanobacterium. This study demonstrates the potential of heterocysts for anaerobic production of biofuels and commodity chemicals under oxygenic photosynthetic conditions.

  1. Relationships between biomass composition and liquid products formed via pyrolysis

    Fan eLin

    2015-10-01

    Full Text Available Thermal conversion of biomass is a rapid, low-cost way to produce a dense liquid product, known as bio-oil, that can be refined to transportation fuels. However, utilization of bio-oil is challenging due to its chemical complexity, acidity, and instability—all results of the intricate nature of biomass. A clear understanding of how biomass properties impact yield and composition of thermal products will provide guidance to optimize both biomass and conditions for thermal conversion. To aid elucidation of these associations, we first describe biomass polymers, including phenolics, polysaccharides, acetyl groups, and inorganic ions, and the chemical interactions among them. We then discuss evidence for three roles (i.e., models for biomass components in formation of liquid pyrolysis products: (1 as direct sources, (2 as catalysts, and (3 as indirect factors whereby chemical interactions among components and/or cell wall structural features impact thermal conversion products. We highlight associations that might be utilized to optimize biomass content prior to pyrolysis, though a more detailed characterization is required to understand indirect effects. In combination with high-throughput biomass characterization techniques this knowledge will enable identification of biomass particularly suited for biofuel production and can also guide genetic engineering of bioenergy crops to improve biomass features.

  2. Relationships between Biomass Composition and Liquid Products Formed via Pyrolysis

    Lin, Fan; Waters, Christopher L.; Mallinson, Richard G.; Lobban, Lance L.; Bartley, Laura E.

    2015-01-01

    Thermal conversion of biomass is a rapid, low-cost way to produce a dense liquid product, known as bio-oil, that can be refined to transportation fuels. However, utilization of bio-oil is challenging due to its chemical complexity, acidity, and instability – all results of the intricate nature of biomass. A clear understanding of how biomass properties impact yield and composition of thermal products will provide guidance to optimize both biomass and conditions for thermal conversion. To aid elucidation of these associations, we first describe biomass polymers, including phenolics, polysaccharides, acetyl groups, and inorganic ions, and the chemical interactions among them. We then discuss evidence for three roles (i.e., models) for biomass components in the formation of liquid pyrolysis products: (1) as direct sources, (2) as catalysts, and (3) as indirect factors whereby chemical interactions among components and/or cell wall structural features impact thermal conversion products. We highlight associations that might be utilized to optimize biomass content prior to pyrolysis, though a more detailed characterization is required to understand indirect effects. In combination with high-throughput biomass characterization techniques, this knowledge will enable identification of biomass particularly suited for biofuel production and can also guide genetic engineering of bioenergy crops to improve biomass features.

  3. Prediction of Gas Leak Tightness of Superplastically Formed Products

    Snippe, Q.H.C.; Meinders, Vincent T.; Barlat, F; Moon, Y.H.; Lee, M.G.

    2010-01-01

    In some applications, in this case an aluminium box in a subatomic particle detector containing highly sensitive detecting devices, it is important that a formed sheet should show no gas leak from one side to the other. In order to prevent a trial-and-error procedure to make this leak tight box, a

  4. Whole-body CO2 production as an index of the metabolic response to sepsis

    Whole-body carbon dioxide (CO2) production (RaCO2) is an index of substrate oxidation and energy expenditure; therefore, it may provide information about the metabolic response to sepsis. Using stable isotope techniques, we determined RaCO2 and its relationship to protein and glucose metabolism in m...

  5. Current floristic composition, life form and productivity of the ...

    SARAH

    2013-01-23

    Jan 23, 2013 ... productivity of the grasslands in the Hunting Zone of. Djona (Benin) ... Moreover, it will be possible to model the impact of grasslands exploitation on the viability of the ... work was run in order to explore native vegetations, current land use ... performed using the flexible beta method based on. Sorensen ...

  6. Immobilisation of radwastes in glass containers and products formed thereby

    Macedo, P.B.; Simmons, C.J.; Lagakos, N.; Simmons, J.H.; Tran, D.C.

    1980-01-01

    A method of preventing the dissemination of toxic material to the environment comprises forming an admixture of toxic material and glass packing in a hollow doped glass container of high silica content, or forming the admixture in a first container and then depositing at least a portion of the admixture in a hollow doped glass container of high silica content. The glass container is then heated to collapse its walls and to seal the container so that the toxic material is entrapped and sealed within the collapsed doped glass container. The thermal expansion coefficient of the container may be decreased prior to use by exchanging hydrogen ion in pores thereof with other cations followed by collapsing the pores. (author)

  7. Metabolic engineering of Agrobacterium sp. ATCC31749 for curdlan production from cellobiose.

    Shin, Hyun-Dong; Liu, Long; Kim, Mi-Kyoung; Park, Yong-Il; Chen, Rachel

    2016-09-01

    Curdlan is a commercial polysaccharide made by fermentation of Agrobacterium sp. Its anticipated expansion to larger volume markets demands improvement in its production efficiency. Metabolic engineering for strain improvement has so far been limited due to the lack of genetic tools. This research aimed to identify strong promoters and to engineer a strain that converts cellobiose efficiently to curdlan. Three strong promoters were identified and were used to install an energy-efficient cellobiose phosphorolysis mechanism in a curdlan-producing strain. The engineered strains were shown with enhanced ability to utilize cellobiose, resulting in a 2.5-fold increase in titer. The availability of metabolically engineered strain capable of producing β-glucan from cellobiose paves the way for its production from cellulose. The identified native promoters from Agrobacterium open up opportunities for further metabolic engineering for improved production of curdlan and other products. The success shown here marks the first such metabolic engineering effort in this microbe.

  8. Metabolic engineering of Ustilago trichophora TZ1 for improved malic acid production

    Thiemo Zambanini

    2017-06-01

    These results open up a wide range of possibilities for further optimization, especially combinatorial metabolic engineering to increase the flux from pyruvate to malic acid and to reduce by-product formation.

  9. A comparative study of zinc protoporphyrin IX-forming properties of animal by-products as sources for improving the color of meat products.

    Wakamatsu, Jun-ichi; Murakami, Naoko; Nishimura, Takanori

    2015-05-01

    The objective of this study was to obtain fundamental data for improving the color of meat products by using animal by-products. We investigated zinc protoporphyrin IX (ZnPP)-forming properties of various internal organs from pigs and chickens. ZnPP was formed in the liver, heart and kidney, whereas the porcine spleen and bile, which are involved in the metabolism of heme, did not have ZnPP-forming properties. The optimum pH values were different among the internal organs and the ZnPP-forming properties of porcine organs were better than those of chicken organs. The porcine liver showed the greatest ZnPP-forming properties among all of the internal organs investigated in this study. The optimum pH value for ZnPP formation in the liver was lower than that of skeletal muscle. Oxygen did not inhibit the formation of ZnPP in the liver, unlike in skeletal muscle. Animal by-products such as the liver have good ability for the formation of ZnPP and might be useful for improving the color of meat products. © 2014 Japanese Society of Animal Science.

  10. Irreducible integrable theories form tensor products of conformal models

    Mathur, S.D.; Warner, N.P.

    1991-01-01

    By using Toda field theories we show that there are perturbations of direct products of conformal theories that lead to irreducible integrable field theories. The same affine Toda theory can be truncated to different quantum integrable models for different choices of the charge at infinity and the coupling. The classification of integrable models that can be obtained in this fashion follows the classification of symmetric spaces of type G/H with rank H = rank G. (orig.)

  11. The product form for path integrals on curved manifolds

    Grosche, C.

    1988-03-01

    A general and simple framework for treating path integrals on curved manifolds is presented. The crucial point will be a product ansatz for the metric tensor and the quantum hamiltonian, i.e. we shall write g αβ = h αγh βγ and H = (1/2m)h αγp αp βh βγ + V + ΔV , respectively, a prescription which we shall call “product form” definition. The p α are hermitian momenta and Δ V is a well-defined quantum correction. We shall show that this ansatz, which looks quite special, is in fact - under reasonable assumptions in quantum mechanics - a very general one. We shall derive the lagrangian path integral in the “product form” definition and shall also prove that the Schro¨dinger equation can be derived from the corresponding short-time kernel. We shall discuss briefly an application of this prescription to the problem of free quantum motion on the Poincare´upper half-plane.

  12. Toward systems metabolic engineering of Aspergillus and Pichia species for the production of chemicals and biofuels

    Caspeta, Luis; Nielsen, Jens

    2013-01-01

    trends in systems biology of Aspergillus and Pichia species, highlighting the relevance of these developments for systems metabolic engineering of these organisms for the production of hydrolytic enzymes, biofuels and chemicals from biomass. Metabolic engineering is moving from traditional methods...... for the production of hydrolytic enzymes, biofuels and chemicals from biomass. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim....

  13. Control of fluxes towards antibiotics and the role of primary metabolism in production of antibiotics

    Gunnarsson, Nina; Eliasson Lantz, Anna; Nielsen, Jacob

    2004-01-01

    Yield improvements in antibiotic-producing strains have classically been obtained through random mutagenesis and screening. An attractive alternative to this strategy is the rational design of producer strains via metabolic engineering, an approach that offers the possibility to increase yields...... in the metabolic network. Here we describe and discuss available methods for identification of these steps, both in antibiotic biosynthesis pathways and in the primary metabolism, which serves as the supplier of precursors and cofactors for the secondary metabolism. Finally, the importance of precursor...... and cofactor supply from primary metabolism in the biosynthesis of different types of antibiotics is discussed and recent developments in metabolic engineering towards increased product yields in antibiotic producing strains are reviewed....

  14. 21 CFR 330.3 - Imprinting of solid oral dosage form drug products.

    2010-04-01

    ... 21 Food and Drugs 5 2010-04-01 2010-04-01 false Imprinting of solid oral dosage form drug products... AS SAFE AND EFFECTIVE AND NOT MISBRANDED General Provisions § 330.3 Imprinting of solid oral dosage form drug products. A requirement to imprint an identification code on solid oral dosage form drug...

  15. A Metabolic Signature of Mitochondrial Dysfunction Revealed through a Monogenic Form of Leigh Syndrome

    Julie Thompson Legault

    2015-11-01

    Full Text Available A decline in mitochondrial respiration represents the root cause of a large number of inborn errors of metabolism. It is also associated with common age-associated diseases and the aging process. To gain insight into the systemic, biochemical consequences of respiratory chain dysfunction, we performed a case-control, prospective metabolic profiling study in a genetically homogenous cohort of patients with Leigh syndrome French Canadian variant, a mitochondrial respiratory chain disease due to loss-of-function mutations in LRPPRC. We discovered 45 plasma and urinary analytes discriminating patients from controls, including classic markers of mitochondrial metabolic dysfunction (lactate and acylcarnitines, as well as unexpected markers of cardiometabolic risk (insulin and adiponectin, amino acid catabolism linked to NADH status (α-hydroxybutyrate, and NAD+ biosynthesis (kynurenine and 3-hydroxyanthranilic acid. Our study identifies systemic, metabolic pathway derangements that can lie downstream of primary mitochondrial lesions, with implications for understanding how the organelle contributes to rare and common diseases.

  16. Metabolic changes during B cell differentiation for the production of intestinal IgA antibody.

    Kunisawa, Jun

    2017-04-01

    To sustain the bio-energetic demands of growth, proliferation, and effector functions, the metabolism of immune cells changes dramatically in response to immunologic stimuli. In this review, I focus on B cell metabolism, especially regarding the production of intestinal IgA antibody. Accumulating evidence has implicated not only host-derived factors (e.g., cytokines) but also gut environmental factors, including the possible involvement of commensal bacteria and diet, in the control of B cell metabolism during intestinal IgA antibody production. These findings yield new insights into the regulation of immunosurveillance and homeostasis in the gut.

  17. Metabolism

    ... lin), which signals cells to increase their anabolic activities. Metabolism is a complicated chemical process, so it's not ... how those enzymes or hormones work. When the metabolism of body chemicals is ... Hyperthyroidism (pronounced: hi-per-THIGH-roy-dih-zum). Hyperthyroidism ...

  18. Meson Form Factors and Deep Exclusive Meson Production Experiments

    Horn, Tanja [The Catholic Univ. of America, Washington, DC (United States); Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States)

    2017-05-01

    Pion and kaon electroproduction data play a unique role in Nature and our understanding of them is essential for explaining hadron structure. Precision longitudinaltransverse separated pion and kaon cross sections are of particular interest. They allow for the extraction of meson form factors and validation of understanding of hard exclusive and semi-inclusive reactions (π+, K+, π0, γ) towards 3D hadron imaging and potential future flavor decomposition. We review recent data and present prospects for deep exclusive pion and kaon electroproduction at the 12 GeV Jefferson Lab including the prospects to use projected charged- and neutral pion data to further determine the spin, charge-parity and flavor of GPDs, including the helicity-flip GPDs.

  19. Methods of forming single source precursors, methods of forming polymeric single source precursors, and single source precursors and intermediate products formed by such methods

    Fox, Robert V.; Rodriguez, Rene G.; Pak, Joshua J.; Sun, Chivin; Margulieux, Kelsey R.; Holland, Andrew W.

    2012-12-04

    Methods of forming single source precursors (SSPs) include forming intermediate products having the empirical formula 1/2{L.sub.2N(.mu.-X).sub.2M'X.sub.2}.sub.2, and reacting MER with the intermediate products to form SSPs of the formula L.sub.2N(.mu.-ER).sub.2M'(ER).sub.2, wherein L is a Lewis base, M is a Group IA atom, N is a Group IB atom, M' is a Group IIIB atom, each E is a Group VIB atom, each X is a Group VIIA atom or a nitrate group, and each R group is an alkyl, aryl, vinyl, (per)fluoro alkyl, (per)fluoro aryl, silane, or carbamato group. Methods of forming polymeric or copolymeric SSPs include reacting at least one of HE.sup.1R.sup.1E.sup.1H and MER with one or more substances having the empirical formula L.sub.2N(.mu.-ER).sub.2M'(ER).sub.2 or L.sub.2N(.mu.-X).sub.2M'(X).sub.2 to form a polymeric or copolymeric SSP. New SSPs and intermediate products are formed by such methods.

  20. Metabolic engineering with systems biology tools to optimize production of prokaryotic secondary metabolites

    Kim, Hyun Uk; Charusanti, Pep; Lee, Sang Yup

    2016-01-01

    Metabolic engineering using systems biology tools is increasingly applied to overproduce secondary metabolites for their potential industrial production. In this Highlight, recent relevant metabolic engineering studies are analyzed with emphasis on host selection and engineering approaches...... for the optimal production of various prokaryotic secondary metabolites: native versus heterologous hosts (e.g., Escherichia coli) and rational versus random approaches. This comparative analysis is followed by discussions on systems biology tools deployed in optimizing the production of secondary metabolites....... The potential contributions of additional systems biology tools are also discussed in the context of current challenges encountered during optimization of secondary metabolite production....

  1. Application of metabolic controls for the maximization of lipid production in semicontinuous fermentation.

    Xu, Jingyang; Liu, Nian; Qiao, Kangjian; Vogg, Sebastian; Stephanopoulos, Gregory

    2017-07-03

    Acetic acid can be generated through syngas fermentation, lignocellulosic biomass degradation, and organic waste anaerobic digestion. Microbial conversion of acetate into triacylglycerols for biofuel production has many advantages, including low-cost or even negative-cost feedstock and environmental benefits. The main issue stems from the dilute nature of acetate produced in such systems, which is costly to be processed on an industrial scale. To tackle this problem, we established an efficient bioprocess for converting dilute acetate into lipids, using the oleaginous yeast Yarrowia lipolytica in a semicontinuous system. The implemented design used low-strength acetic acid in both salt and acid forms as carbon substrate and a cross-filtration module for cell recycling. Feed controls for acetic acid and nitrogen based on metabolic models and online measurement of the respiratory quotient were used. The optimized process was able to sustain high-density cell culture using acetic acid of only 3% and achieved a lipid titer, yield, and productivity of 115 g/L, 0.16 g/g, and 0.8 g⋅L -1 ⋅h -1 , respectively. No carbon substrate was detected in the effluent stream, indicating complete utilization of acetate. These results represent a more than twofold increase in lipid production metrics compared with the current best-performing results using concentrated acetic acid as carbon feed.

  2. Metabolic Engineering of TCA Cycle for Production of Chemicals

    Vuoristo, K.S.; Mars, A.E.; Sanders, J.P.M.; Eggink, G.; Weusthuis, R.A.

    2016-01-01

    The tricarboxylic acid (TCA) cycle has been used for decades in the microbial production of chemicals such as citrate, L-glutamate, and succinate. Maximizing yield is key for cost-competitive production. However, for most TCA cycle products, the maximum pathway yield is lower than the theoretical

  3. From pathways to genomes and beyond. The metabolic engineering toolbox and its place in biofuels production

    Liu, Leqian; Reed, Ben; Alper, Hal [Texas Univ., Austin, TX (United States). Dept. of Chemical Engineering

    2011-07-01

    Concerns about the availability of petroleum-derived fuels and chemicals have led to the exploration of metabolically engineered organisms as novel hosts for biofuels and chemicals production. However, the complexity inherent in metabolic and regulatory networks makes this undertaking a complex task. To address these limitations, metabolic engineering has adapted a wide-variety of tools for altering phenotypes. In this review, we will highlight traditional and recent metabolic engineering tools for optimizing cells including pathway-based, global, and genomic-enabled approaches. Specifically, we describe these tools as well as provide demonstrations of their effectiveness in optimizing biofuels production. However, each of these tools provides stepping stones towards the grand goal of biofuels production. Thus, developing methods for large-scale cellular optimization and integrative approaches are invaluable for further cell optimization. This review highlights the challenges that still must be met to accomplish this goal. (orig.)

  4. Identification and quantification of phencyclidine pyrolysis products formed during smoking.

    Lue, L P; Scimeca, J A; Thomas, B F; Martin, B R

    1986-01-01

    As a result of frequent phencyclidine (PCP) abuse, pyrolysis studies were conducted to further investigate its fate during smoking. Marijuana placebo cigarettes were impregnated with 3H-PCP X HCI and burned under conditions simulating smoking. Mainstream smoke was passed through glass wool filters as well as acidic and basic traps. Approximately 90% of the starting material could be accounted for in the first glass wool trap and cigarette holder. HPLC and GC/MS analysis of methanol extracts of these glass wool traps revealed the presence of 1-phenyl-1-cyclohexene (47% of the starting material) greater than PCP (40%) greater than piperidine (15%) greater than N-acetylpiperidine (9%). It was not possible to fully account for the remainder of the piperidine moiety. It has been reported that at high temperatures PCP is converted to numerous polynuclear aromatic compounds which include styrene, alpha-methylstyrene, naphthalene, 2-methylnaphthalene, 1-methylnaphthalene, biphenyl, cyclohexylbenzene, acenaphthene, phenanthrene, and anthracene. These compounds were not formed from PCP under smoking conditions.

  5. Identification and quantification of phencyclidine pyrolysis products formed during smoking

    Lue, L.P.; Scimeca, J.A.; Thomas, B.F.; Martin, B.R.

    1986-01-01

    As a result of frequent phencyclidine (PCP) abuse, pyrolysis studies were conducted to further investigate its fate during smoking. Marijuana placebo cigarettes were impregnated with 3 H-PCP HCl and burned under conditions simulating smoking. Mainstream smoke was passed through glass wool filters as well as acidic and basic traps. Approximately 90% of the starting material could be accounted for in the first glass wool trap and cigarette holder. HPLC and GC/MS analysis of methanol extracts of these glass wool traps revealed the presence of 1-phenyl-1-cyclohexene (47% of the starting material) > PCP (40%) > piperidine (15%) > N-acetylpiperidine (9%). It was not possible to fully account for the remainder of the piperidine moiety. It has been reported that at high temperatures PCP is converted to numerous polynuclear aromatic compounds which include styrene, α-methylstyrene, naphthalene, 2-methyl-naphthalene, 1-methylnaphthalene, biphenyl, cyclohexylbenzene, acenaphthene, phenanthrene, and anthracene. These compounds were not formed from PCP under smoking conditions

  6. Improved Triacylglycerol Production in Acinetobacter baylyi ADP1 by Metabolic Engineering

    Karp Matti

    2011-05-01

    Full Text Available Abstract Background Triacylglycerols are used in various purposes including food applications, cosmetics, oleochemicals and biofuels. Currently the main sources for triacylglycerol are vegetable oils, and microbial triacylglycerol has been suggested as an alternative for these. Due to the low production rates and yields of microbial processes, the role of metabolic engineering has become more significant. As a robust model organism for genetic and metabolic studies, and for the natural capability to produce triacylglycerol, Acinetobacter baylyi ADP1 serves as an excellent organism for modelling the effects of metabolic engineering for energy molecule biosynthesis. Results Beneficial gene deletions regarding triacylglycerol production were screened by computational means exploiting the metabolic model of ADP1. Four deletions, acr1, poxB, dgkA, and a triacylglycerol lipase were chosen to be studied experimentally both separately and concurrently by constructing a knock-out strain (MT with three of the deletions. Improvements in triacylglycerol production were observed: the strain MT produced 5.6 fold more triacylglycerol (mg/g cell dry weight compared to the wild type strain, and the proportion of triacylglycerol in total lipids was increased by 8-fold. Conclusions In silico predictions of beneficial gene deletions were verified experimentally. The chosen single and multiple gene deletions affected beneficially the natural triacylglycerol metabolism of A. baylyi ADP1. This study demonstrates the importance of single gene deletions in triacylglycerol metabolism, and proposes Acinetobacter sp. ADP1 as a model system for bioenergetic studies regarding metabolic engineering.

  7. The Genome-Based Metabolic Systems Engineering to Boost Levan Production in a Halophilic Bacterial Model.

    Aydin, Busra; Ozer, Tugba; Oner, Ebru Toksoy; Arga, Kazim Yalcin

    2018-03-01

    Metabolic systems engineering is being used to redirect microbial metabolism for the overproduction of chemicals of interest with the aim of transforming microbial hosts into cellular factories. In this study, a genome-based metabolic systems engineering approach was designed and performed to improve biopolymer biosynthesis capability of a moderately halophilic bacterium Halomonas smyrnensis AAD6 T producing levan, which is a fructose homopolymer with many potential uses in various industries and medicine. For this purpose, the genome-scale metabolic model for AAD6 T was used to characterize the metabolic resource allocation, specifically to design metabolic engineering strategies for engineered bacteria with enhanced levan production capability. Simulations were performed in silico to determine optimal gene knockout strategies to develop new strains with enhanced levan production capability. The majority of the gene knockout strategies emphasized the vital role of the fructose uptake mechanism, and pointed out the fructose-specific phosphotransferase system (PTS fru ) as the most promising target for further metabolic engineering studies. Therefore, the PTS fru of AAD6 T was restructured with insertional mutagenesis and triparental mating techniques to construct a novel, engineered H. smyrnensis strain, BMA14. Fermentation experiments were carried out to demonstrate the high efficiency of the mutant strain BMA14 in terms of final levan concentration, sucrose consumption rate, and sucrose conversion efficiency, when compared to the AAD6 T . The genome-based metabolic systems engineering approach presented in this study might be considered an efficient framework to redirect microbial metabolism for the overproduction of chemicals of interest, and the novel strain BMA14 might be considered a potential microbial cell factory for further studies aimed to design levan production processes with lower production costs.

  8. Metabolically engineered cells for the production of polyunsaturated fatty acids

    2005-01-01

    The present invention relates to the construction and engineering of cells, more particularly microorganisms for producing PUFAs with four or more double bonds from non-fatty acid substrates through heterologous expression of an oxygen requiring pathway. The invention especially involves...... improvement of the PUFA content in the host organism through fermentation optimization, e.g. decreasing the temperature and/or designing an optimal medium, or through improving the flux towards fatty acids by metabolic engineering, e.g. through over-expression of fatty acid synthases, over-expression of other...

  9. Oxidation of dibenzothiophene (DBT by Serratia marcescens UCP 1549 formed biphenyl as final product

    de Araújo Hélvia W

    2012-05-01

    Full Text Available Abstract Background The desulphurization of dibenzothiophene (DBT, a recalcitrant thiophenic fossil fuel component by Serratia marcescens (UCP 1549 in order for reducing the Sulphur content was investigated. The Study was carried out establishing the growth profile using Luria Bertani medium to different concentrations of DBT during 120 hours at 28°C, and orbital Shaker at 150 rpm. Results The results indicated that concentrations of DBT 0.5, 1.0 and 2.0 mM do not affected the growth of the bacterium. The DBT showed similar Minimum Inhibitory Concentration (MIC and Minimum Bactericidal Concentration (MCB (3.68 mM. The desulphurization of DBT by S. marcescens was used with 96 hours of growth on 2 mM of DBT, and was determined by gas chromatography (GC and GC-mass spectrometry. In order to study the desulphurization process by S. marcescens was observed the presence of a sulfur-free product at 16 hours of cultivation. Conclusions The data suggests the use of metabolic pathway “4S” by S. marcescens (UCP 1549 and formed biphenyl. The microbial desulphurization process by Serratia can be suggest significant reducing sulphur content in DBT, and showed promising potential for reduction of the sulfur content in diesel oil.

  10. Proteolytic receptor cleavage in the pathogenesis of blood rheology and co-morbidities in metabolic syndrome. Early forms of autodigestion.

    Mazor, Rafi; Schmid-Schönbein, Geert W

    2015-01-01

    Abnormal blood rheological properties seldom occur in isolation and instead are accompanied by other complications, often designated as co-morbidities. In the metabolic syndrome with complications like hypertension, diabetes and lack of normal microvascular blood flow, the underlying molecular mechanisms that simultaneously lead to elevated blood pressure and diabetes as well as abnormal microvascular rheology and other cell dysfunctions have remained largely unknown. In this review, we propose a new hypothesis for the origin of abnormal cell functions as well as multiple co-morbidities. Utilizing experimental models for the metabolic disease with diverse co-morbidities we summarize evidence for the presence of an uncontrolled extracellular proteolytic activity that causes ectodomain receptor cleavage and loss of their associated cell function. We summarize evidence for unchecked degrading proteinase activity, e.g. due to matrix metalloproteases, in patients with hypertension, Type II diabetes and obesity, in addition to evidence for receptor cleavage in the form of receptor fragments and decreased extracellular membrane expression levels. The evidence suggest that a shift in blood rheological properties and other co-morbidities may in fact be derived from a common mechanism that is due to uncontrolled proteolytic activity, i.e. an early form of autodigestion. Identification of the particular proteases involved and the mechanisms of their activation may open the door to treatment that simultaneously targets multiple co-morbidities in the metabolic syndrome.

  11. Biodegradation of Cosmetics Products: A Computational Study of Cytochrome P450 Metabolism of Phthalates

    Fabián G. Cantú Reinhard

    2017-11-01

    Full Text Available Cytochrome P450s are a broad class of enzymes in the human body with important functions for human health, which include the metabolism and detoxification of compounds in the liver. Thus, in their catalytic cycle, the P450s form a high-valent iron(IV-oxo heme cation radical as the active species (called Compound I that reacts with substrates through oxygen atom transfer. This work discusses the possible degradation mechanisms of phthalates by cytochrome P450s in the liver, through computational modelling, using 2-ethylhexyl-phthalate as a model substrate. Phthalates are a type of compound commonly found in the environment from cosmetics usage, but their biodegradation in the liver may lead to toxic metabolites. Experimental studies revealed a multitude of products and varying product distributions among P450 isozymes. To understand the regio- and chemoselectivity of phthalate activation by P450 isozymes, we focus here on the mechanisms of phthalate activation by Compound I leading to O-dealkylation, aliphatic hydroxylation and aromatic hydroxylation processes. We set up model complexes of Compound I with the substrate and investigated the reaction mechanisms for products using the density functional theory on models and did a molecular mechanics study on enzymatic structures. The work shows that several reaction barriers in the gas-phase are close in energy, leading to a mixture of products. However, when we tried to dock the substrate into a P450 isozyme, some of the channels were inaccessible due to unfavorable substrate positions. Product distributions are discussed under various reaction conditions and rationalized with valence bond and thermodynamic models.

  12. Metabolic products of linalool and modulation of GABAA receptors

    Milanos, Sinem; Elsharif, Shaimaa A.; Janzen, Dieter; Buettner, Andrea; Villmann, Carmen

    2017-06-01

    Terpenoids are major subcomponents in aroma substances which harbor sedative physiological potential. We have demonstrated that various monoterpenoids such as the acyclic linalool enhance GABAergic currents in an allosteric manner in vitro upon overexpression of inhibitory a1b2 GABAA receptors in various expression systems. However, in plants or humans, i.e. following intake via inhalation or ingestion, linalool undergoes metabolic modifications including oxygenation and acetylation, which may affect the modulatory efficacy of the generated linalool derivatives. Here, we analyzed the modulatory potential of linalool derivatives at a1b2g2 GABAA receptors upon transient overexpression. Following receptor expression control, electrophysiological recordings in a whole cell configuration were used to determine the chloride influx upon co-application of GABA EC5-10 together with the modulatory substance. Our results show that only oxygenated linalool metabolites at carbon 8 positively affect GABAergic currents whereas derivatives hydroxylated or carboxylated at carbon 8 were rather ineffective. Acetylated linalool derivatives resulted in non-significant changes of GABAergic currents. We can conclude that metabolism of linalool reduces its positive allosteric potential at GABAA receptors compared to the significant potentiation effects of the parent molecule linalool itself.

  13. Metabolic and environmental aspects of fusion reactor activation products: niobium

    Easterly, C.E.; Shank, K.E.

    1977-11-01

    A summary of the metabolic and environmental aspects of niobium is presented. The toxicological symptoms from exposure to niobium are given, along with lethal concentration values for acute and chronic exposures. Existing human data are presented; animal uptake and retention data are analyzed for various routes of administration. Recommended metabolic values are also presented along with comments concerning their use and appropriateness. The natural distribution of niobium is given for freshwater, seawater, and the biosphere. Concentration factors and retention of 95 Nb in the environment are discussed with reference to: plant retention via leaf absorption; plant retention via root uptake; uptake in terrestrial animals from plants; uptake in freshwater organisms; uptake in marine organisms; and movement in soil. Conclusions are drawn regarding needs for future work in these areas. This review was undertaken because niobium is expected to be a key metal in the development of commercial fusion reactors. It is recognized that niobium will likely not be used in the first generation reactors as a structural material but will appear as an alloy in such materials as superconducting wire

  14. Metabolic Products of Linalool and Modulation of GABAA Receptors

    Sinem Milanos

    2017-06-01

    Full Text Available Terpenoids are major subcomponents in aroma substances which harbor sedative physiological potential. We have demonstrated that various monoterpenoids such as the acyclic linalool enhance GABAergic currents in an allosteric manner in vitro upon overexpression of inhibitory α1β2 GABAA receptors in various expression systems. However, in plants or humans, i.e., following intake via inhalation or ingestion, linalool undergoes metabolic modifications including oxygenation and acetylation, which may affect the modulatory efficacy of the generated linalool derivatives. Here, we analyzed the modulatory potential of linalool derivatives at α1β2γ2 GABAA receptors upon transient overexpression. Following receptor expression control, electrophysiological recordings in a whole cell configuration were used to determine the chloride influx upon co-application of GABA EC10−30 together with the modulatory substance. Our results show that only oxygenated linalool metabolites at carbon 8 positively affect GABAergic currents whereas derivatives hydroxylated or carboxylated at carbon 8 were rather ineffective. Acetylated linalool derivatives resulted in non-significant changes of GABAergic currents. We can conclude that metabolism of linalool reduces its positive allosteric potential at GABAA receptors compared to the significant potentiation effects of the parent molecule linalool itself.

  15. Metabolic and environmental aspects of fusion reactor activation products: niobium

    Easterly, C.E.; Shank, K.E.

    1977-11-01

    A summary of the metabolic and environmental aspects of niobium is presented. The toxicological symptoms from exposure to niobium are given, along with lethal concentration values for acute and chronic exposures. Existing human data are presented; animal uptake and retention data are analyzed for various routes of administration. Recommended metabolic values are also presented along with comments concerning their use and appropriateness. The natural distribution of niobium is given for freshwater, seawater, and the biosphere. Concentration factors and retention of /sup 95/Nb in the environment are discussed with reference to: plant retention via leaf absorption; plant retention via root uptake; uptake in terrestrial animals from plants; uptake in freshwater organisms; uptake in marine organisms; and movement in soil. Conclusions are drawn regarding needs for future work in these areas. This review was undertaken because niobium is expected to be a key metal in the development of commercial fusion reactors. It is recognized that niobium will likely not be used in the first generation reactors as a structural material but will appear as an alloy in such materials as superconducting wire.

  16. Fructose increases corticosterone production in association with NADPH metabolism alterations in rat epididymal white adipose tissue.

    Prince, Paula D; Santander, Yanina A; Gerez, Estefania M; Höcht, Christian; Polizio, Ariel H; Mayer, Marcos A; Taira, Carlos A; Fraga, Cesar G; Galleano, Monica; Carranza, Andrea

    2017-08-01

    Metabolic syndrome is an array of closely metabolic disorders that includes glucose intolerance/insulin resistance, central obesity, dyslipidemia, and hypertension. Fructose, a highly lipogenic sugar, has profound metabolic effects in adipose tissue, and has been associated with the etiopathology of many components of the metabolic syndrome. In adipocytes, the enzyme 11 β-HSD1 amplifies local glucocorticoid production, being a key player in the pathogenesis of central obesity and metabolic syndrome. 11 β-HSD1 reductase activity is dependent on NADPH, a cofactor generated by H6PD inside the endoplasmic reticulum. Our focus was to explore the effect of fructose overload on epididymal white adipose tissue (EWAT) machinery involved in glucocorticoid production and NADPH and oxidants metabolism. Male Sprague-Dawley rats fed with a fructose solution (10% (w/v) in tap water) during 9 weeks developed some characteristic features of metabolic syndrome, such as hypertriglyceridemia, and hypertension. In addition, high levels of plasma and EWAT corticosterone were detected. Activities and expressions of H6PD and 11 β-HSD1, NAPDH content, superoxide anion production, expression of NADPH oxidase 2 subunits, and indicators of oxidative metabolism were measured. Fructose overloaded rats showed an increased potential in oxidant production respect to control rats. In parallel, in EWAT from fructose overloaded rats we found higher expression/activity of H6PD and 11 β-HSD1, and NADPH/NADP + ratio. Our in vivo results support that fructose overload installs in EWAT conditions favoring glucocorticoid production through higher H6PD expression/activity supplying NADPH for enhanced 11 β-HSD1 expression/activity, becoming this tissue a potential extra-adrenal source of corticosterone under these experimental conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Lipid and fatty acid metabolism in Ralstonia eutropha: relevance for the biotechnological production of value-added products.

    Riedel, Sebastian L; Lu, Jingnan; Stahl, Ulf; Brigham, Christopher J

    2014-02-01

    Lipid and fatty acid metabolism has been well studied in model microbial organisms like Escherichia coli and Bacillus subtilis. The major precursor of fatty acid biosynthesis is also the major product of fatty acid degradation (β-oxidation), acetyl-CoA, which is a key metabolite for all organisms. Controlling carbon flux to fatty acid biosynthesis and from β-oxidation allows for the biosynthesis of natural products of biotechnological importance. Ralstonia eutropha can utilize acetyl-CoA from fatty acid metabolism to produce intracellular polyhydroxyalkanoate (PHA). R. eutropha can also be engineered to utilize fatty acid metabolism intermediates to produce different PHA precursors. Metabolism of lipids and fatty acids can be rerouted to convert carbon into other value-added compounds like biofuels. This review discusses the lipid and fatty acid metabolic pathways in R. eutropha and how they can be used to construct reagents for the biosynthesis of products of industrial importance. Specifically, how the use of lipids or fatty acids as the sole carbon source in R. eutropha cultures adds value to these biotechnological products will be discussed here.

  18. Metabolic engineering of Escherichia coli for itaconate production

    Vuoristo, K.S.

    2016-01-01

    Interest in sustainable development together with limited amounts of fossil resources have increased the demand for production of chemicals and fuels from renewable resources. The market potential for bio-based products is growing and a transition from petrochemicals to biomass-based chemicals is

  19. Metabolic engineering of Saccharomyces cerevisiae for optimizing 3HP production

    Jensen, Niels Bjerg; Maury, Jerome; Oberg, Fredrik

    2012-01-01

    . Polyacrylates are a substantial part of the different plastic varieties found on the market. This kind of plastic is derived from acrylic acid, which is currently produced from propylene, a by-product of ethylene and gasoline production. Annually, more than one billion kilograms of acrylic acid is produced...

  20. L-malate production by metabolically engineered escherichia coli

    Zhang, Xueli; Wang, Xuan; Shanmugam, Keelnatham T.; Ingram, Lonnie O'Neal

    2015-11-17

    A process for the production of malic acid in commercially significant quantities from the carbon compounds by genetically modified bacterial strains (GMBS; also referred to as biocatalysts or genetically modified microorganisms) is disclosed. Microorganisms suitable for the production of malic acid can be cultured in one or two-step processes as disclosed herein.

  1. Metabolic Engineering of TCA Cycle for Production of Chemicals.

    Vuoristo, Kiira S; Mars, Astrid E; Sanders, Johan P M; Eggink, Gerrit; Weusthuis, Ruud A

    2016-03-01

    The tricarboxylic acid (TCA) cycle has been used for decades in the microbial production of chemicals such as citrate, L-glutamate, and succinate. Maximizing yield is key for cost-competitive production. However, for most TCA cycle products, the maximum pathway yield is lower than the theoretical maximum yield (Y(E)). For succinate, this was solved by creating two pathways to the product, using both branches of the TCA cycle, connected by the glyoxylate shunt (GS). A similar solution cannot be applied directly for production of compounds from the oxidative branch of the TCA cycle because irreversible reactions are involved. Here, we describe how this can be overcome and what the impact is on the yield. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Metabolism of skin-absorbed resveratrol into its glucuronized form in mouse skin.

    Itsuo Murakami

    Full Text Available Resveratrol (RESV is a plant polyphenol, which is thought to have beneficial metabolic effects in laboratory animals as well as in humans. Following oral administration, RESV is immediately catabolized, resulting in low bioavailability. This study compared RESV metabolites and their tissue distribution after oral uptake and skin absorption. Metabolomic analysis of various mouse tissues revealed that RESV can be absorbed and metabolized through skin. We detected sulfated and glucuronidated RESV metabolites, as well as dihydroresveratrol. These metabolites are thought to have lower pharmacological activity than RESV. Similar quantities of most RESV metabolites were observed 4 h after oral or skin administration, except that glucuronidated RESV metabolites were more abundant in skin after topical RESV application than after oral administration. This result is consistent with our finding of glucuronidated RESV metabolites in cultured skin cells. RESV applied to mouse ears significantly suppressed inflammation in the TPA inflammation model. The skin absorption route could be a complementary, potent way to achieve therapeutic effects with RESV.

  3. Metabolic production of amphetamine following multidose administration of clobenzorex.

    Baden, K L; Valtier, S; Cody, J T

    1999-10-01

    The interpretation of urine drug-testing results can have important forensic and legal implications. In particular, drugs that are metabolized to amphetamine or methamphetamine or both pose significant concerns. In this study, clobenzorex, an anorectic drug that is metabolized to d-amphetamine, was administered to five subjects. Each subject took 30 mg daily for seven days, and individual urine samples were collected ad lib for 14 days beginning on the first day the drug was administered. Urine pH, specific gravity, and creatinine values were determined for each sample. Gas chromatography-mass spectrometry (GC-MS) was used to determine the excretion profile of amphetamine and clobenzorex using a standard procedure for amphetamines with additional monitoring of ions at m/z 118, 125, and 364 for the detection of clobenzorex. Peak concentrations of amphetamine were found at 82 to 168 h after the first dose and ranged from approximately 2900 to 4700 ng/mL amphetamine. The use of a regioisomer (3-Cl-benzylamphetamine) as internal standard allowed for accurate quantitation of the parent drug. Peak concentrations of clobenzorex were found at 50 to 120 h after the first dose and ranged from approximately 8 to 47 ng/mL clobenzorex. However, in many samples, clobenzorex was not detected at all. This analysis revealed that the metabolite, (amphetamine) is present in much higher concentrations than the parent compound, clobenzorex. Yet even at peak amphetamine concentrations, the parent was not always detected (limit of detection 1 ng/mL). Thus, in the interpretation of amphetamine-positive drug-testing results, the absence of clobenzorex in the urine sample does not exclude the possibility of its use.

  4. Dedicated Industrial Oilseed Crops as Metabolic Engineering Platforms for Sustainable Industrial Feedstock Production.

    Zhu, Li-Hua; Krens, Frans; Smith, Mark A; Li, Xueyuan; Qi, Weicong; van Loo, Eibertus N; Iven, Tim; Feussner, Ivo; Nazarenus, Tara J; Huai, Dongxin; Taylor, David C; Zhou, Xue-Rong; Green, Allan G; Shockey, Jay; Klasson, K Thomas; Mullen, Robert T; Huang, Bangquan; Dyer, John M; Cahoon, Edgar B

    2016-02-26

    Feedstocks for industrial applications ranging from polymers to lubricants are largely derived from petroleum, a non-renewable resource. Vegetable oils with fatty acid structures and storage forms tailored for specific industrial uses offer renewable and potentially sustainable sources of petrochemical-type functionalities. A wide array of industrial vegetable oils can be generated through biotechnology, but will likely require non-commodity oilseed platforms dedicated to specialty oil production for commercial acceptance. Here we show the feasibility of three Brassicaceae oilseeds crambe, camelina, and carinata, none of which are widely cultivated for food use, as hosts for complex metabolic engineering of wax esters for lubricant applications. Lines producing wax esters >20% of total seed oil were generated for each crop and further improved for high temperature oxidative stability by down-regulation of fatty acid polyunsaturation. Field cultivation of optimized wax ester-producing crambe demonstrated commercial utility of these engineered crops and a path for sustainable production of other industrial oils in dedicated specialty oilseeds.

  5. Brain Glucose Metabolism Controls Hepatic Glucose and Lipid Production

    Lam, Tony K.T.

    2007-01-01

    Brain glucose-sensing mechanisms are implicated in the regulation of feeding behavior and hypoglycemic-induced hormonal counter-regulation. This commentary discusses recent findings indicating that the brain senses glucose to regulate both hepatic glucose and lipid production.

  6. Water deficit affects primary metabolism differently in two Lolium multiflorum/Festuca arundinacea introgression forms with a distinct capacity for photosynthesis and membrane regeneration.

    Dawid Perlikowski

    2016-07-01

    Full Text Available Understanding how plants respond to drought at different levels of cell metabolism is an important aspect of research on the mechanisms involved in stress tolerance. Furthermore, a dissection of drought tolerance into its crucial components by the use of plant introgression forms facilitates to analyze this trait more deeply. The important components of plant drought tolerance are the capacity for photosynthesis under drought conditions, and the ability of cellular membrane regeneration after stress cessation. Two closely related introgression forms of Lolium multiflorum/Festuca arundinacea, differing in the level of photosynthetic capacity during stress, and in the ability to regenerate their cellular membranes after stress cessation, were used as forage grass models in a primary metabolome profiling and in an evaluation of chloroplast 1,6-bisphosphate aldolase accumulation level and activity, during 11 days of water deficit, followed by 10 days of rehydration. It was revealed here that the introgression form, characterized by the ability to regenerate membranes after rehydration, contained higher amounts of proline, melibiose, galactaric acid, myo-inositol and myo-inositol-1-phosphate involved in osmoprotection and stress signaling under drought. Moreover, during the rehydration period, this form also maintained elevated accumulation levels of most the primary metabolites, analyzed here. The other introgression form, characterized by the higher capacity for photosynthesis, revealed a higher accumulation level and activity of chloroplast aldolase under drought conditions, and higher accumulation levels of most photosynthetic products during control and drought periods. The potential impact of the observed metabolic alterations on cellular membrane recovery after stress cessation, and on a photosynthetic capacity under drought conditions in grasses, are discussed.

  7. Systems metabolic engineering design: fatty acid production as an emerging case study.

    Tee, Ting Wei; Chowdhury, Anupam; Maranas, Costas D; Shanks, Jacqueline V

    2014-05-01

    Increasing demand for petroleum has stimulated industry to develop sustainable production of chemicals and biofuels using microbial cell factories. Fatty acids of chain lengths from C6 to C16 are propitious intermediates for the catalytic synthesis of industrial chemicals and diesel-like biofuels. The abundance of genetic information available for Escherichia coli and specifically, fatty acid metabolism in E. coli, supports this bacterium as a promising host for engineering a biocatalyst for the microbial production of fatty acids. Recent successes rooted in different features of systems metabolic engineering in the strain design of high-yielding medium chain fatty acid producing E. coli strains provide an emerging case study of design methods for effective strain design. Classical metabolic engineering and synthetic biology approaches enabled different and distinct design paths towards a high-yielding strain. Here we highlight a rational strain design process in systems biology, an integrated computational and experimental approach for carboxylic acid production, as an alternative method. Additional challenges inherent in achieving an optimal strain for commercialization of medium chain-length fatty acids will likely require a collection of strategies from systems metabolic engineering. Not only will the continued advancement in systems metabolic engineering result in these highly productive strains more quickly, this knowledge will extend more rapidly the carboxylic acid platform to the microbial production of carboxylic acids with alternate chain-lengths and functionalities. © 2014 Wiley Periodicals, Inc.

  8. The prevalence of metabolic syndrome in an employed population and the impact on health and productivity.

    Burton, Wayne N; Chen, Chin-Yu; Schultz, Alyssa B; Edington, Dee W

    2008-10-01

    To investigate the prevalence of metabolic syndrome in an employed population and its association with health risks, health perception, illness days, work limitation (presenteeism), and short-term disability (STD). Five thousand five hundred twelve employees of a financial services company responded to an on-site health risk appraisal which included measured waist circumference and biometric results. The metabolic syndrome criteria were based on the 2005 AHA/NHLBI scientific statement on the diagnosis and management of metabolic syndrome. Perceived health, illness days, and presenteeism were self-reported; STD days were obtained from claims data. In this employee population (61% women, average age 41 years), 22.6% met the criteria for metabolic syndrome and were more likely to report more health risks, poorer health perception, and more absent days due to illness. There was no clear association with presenteeism or STD incidence. However, as the number of metabolic risk factors increased, there was an increase in STD incidence, decrease in health perception, and increase in illness days. No association was found with number of metabolic risk factors and presenteeism. Metabolic syndrome was associated with poor perceived health, increased illness days, and an increased trend of STD incidence. Worksite health promotion programs could be useful in helping employees and employers to identify metabolic syndrome risks and take steps to reduce risk and potential productivity losses.

  9. Metabolic differentiation in biofilms as indicated by carbon dioxide production rates.

    Bester, Elanna; Kroukamp, Otini; Wolfaardt, Gideon M; Boonzaaier, Leandro; Liss, Steven N

    2010-02-01

    The measurement of carbon dioxide production rates as an indication of metabolic activity was applied to study biofilm development and response of Pseudomonas sp. biofilms to an environmental disturbance in the form of a moving air-liquid interface (i.e., shear). A differential response in biofilm cohesiveness was observed after bubble perturbation, and the biofilm layers were operationally defined as either shear-susceptible or non-shear-susceptible. Confocal laser scanning microscopy and image analysis showed a significant reduction in biofilm thickness and biomass after the removal of the shear-susceptible biofilm layer, as well as notable changes in the roughness coefficient and surface-to-biovolume ratio. These changes were accompanied by a 72% reduction of whole-biofilm CO2 production; however, the non-shear-susceptible region of the biofilm responded rapidly after the removal of the overlying cells and extracellular polymeric substances (EPS) along with the associated changes in nutrient and O2 flux, with CO2 production rates returning to preperturbation levels within 24 h. The adaptable nature and the ability of bacteria to respond to environmental conditions were further demonstrated by the outer shear-susceptible region of the biofilm; the average CO2 production rate of cells from this region increased within 0.25 h from 9.45 +/- 5.40 fmol of CO2 x cell(-1) x h(-1) to 22.6 +/- 7.58 fmol of CO2 x cell(-1) x h(-1) when cells were removed from the biofilm and maintained in suspension without an additional nutrient supply. These results also demonstrate the need for sufficient monitoring of biofilm recovery at the solid substratum if mechanical methods are used for biofouling control.

  10. Plutonium and surrogate fission products in a composite ceramic waste form

    Esh, D. W.; Frank, S. M.; Goff, K. M.; Johnson, S. G.; Moschetti, T. L.; O'Holleran, T.

    1999-01-01

    Argonne National Laboratory is developing a ceramic waste form to immobilize salt containing fission products and transuranic elements. Preliminary results have been presented for ceramic waste forms containing surrogate fission products such as cesium and the lanthanides. In this work results from scanning electron microscopy/energy dispersive spectroscopy and x-ray diffraction are presented in greater detail for ceramic waste forms containing surrogate fission products. Additionally, results for waste forms containing plutonium and surrogate fission products are presented. Most of the surrogate fission products appear to be silicates or aluminosilicates whereas the plutonium is usually found in an oxide form. There is also evidence for the presence of plutonium within the sodalite phase although the chemical speciation of the plutonium is not known

  11. Forms and Factors of Animal Products Marketing in the early post-Meiji Years

    Ono, Seishi

    1982-01-01

    As far as the period after the Meiji era is concerned, at its begining marketing of animal products was formed in conlpliance with the properties of products as a public commodity and the amount of investment needed. The former factor depends basically on the divisibility and the preservation possibilities of the products. The latter is the capital needed for marketing and production. 1) Eggs are the most divisible and preservable of all the animal products. Also, hens can be kept with sm...

  12. Comparative study of hops-containing products on human cytochrome P450-mediated metabolism.

    Foster, Brian C; Arnason, John T; Saleem, Ammar; Tam, Teresa W; Liu, Rui; Mao, Jingqin; Desjardins, Suzanne

    2011-05-11

    The potential for 15 different ales (6), ciders (2 apple and 1 pear), and porters (6) and 2 non-alcoholic products to affect cytochrome P450 (CYP)-mediated biotransformation and P-glycoprotein-mediated efflux of rhodamine was examined. As in our previous study, a wide range of recovered nonvolatile suspended solids dry weights were noted. Aliquots were also found to have varying effects on biotransformation and efflux. Distinct differences in product ability to affect the safety and efficacy of therapeutic products confirmed our initial findings that some porters (stouts) have a potential to affect the safety and efficacy of health products metabolized by CYP2D6 and CYP3A4 isozymes. Most products, except 2 of the ciders and the 2 non-alcoholic products, also have the potential to affect the safety of CYP2C9 metabolized medications and supplements. Further studies are required to determine the clinical significance of these findings.

  13. Metabolic Engineering for Production of Biorenewable Fuels and Chemicals: Contributions of Synthetic Biology

    Laura R. Jarboe

    2010-01-01

    Full Text Available Production of fuels and chemicals through microbial fermentation of plant material is a desirable alternative to petrochemical-based production. Fermentative production of biorenewable fuels and chemicals requires the engineering of biocatalysts that can quickly and efficiently convert sugars to target products at a cost that is competitive with existing petrochemical-based processes. It is also important that biocatalysts be robust to extreme fermentation conditions, biomass-derived inhibitors, and their target products. Traditional metabolic engineering has made great advances in this area, but synthetic biology has contributed and will continue to contribute to this field, particularly with next-generation biofuels. This work reviews the use of metabolic engineering and synthetic biology in biocatalyst engineering for biorenewable fuels and chemicals production, such as ethanol, butanol, acetate, lactate, succinate, alanine, and xylitol. We also examine the existing challenges in this area and discuss strategies for improving biocatalyst tolerance to chemical inhibitors.

  14. Characterization of corrosion products formed on steels in the first months of atmospheric exposure

    Antunes Renato Altobelli; Costa Isolda; Faria Dalva Lúcia Araújo de

    2003-01-01

    The corrosion products of carbon steel and weathering steel exposed to three different types of atmospheres, at times ranging from one to three months, have been identified. The steels were exposed in an industrial site, an urban site (São Paulo City, Brazil), and a humid site. The effect of the steel type on the corrosion products formed in the early stages of atmospheric corrosion has been evaluated. The corrosion products formed at the various exposure locations were characterized by Raman...

  15. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

    Fleige, Christian; Meyer, Florian; Steinbüchel, Alexander

    2016-01-01

    The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for ...

  16. Regulation of metabolic products and gene expression in Fusarium asiaticum by agmatine addition.

    Suzuki, Tadahiro; Kim, Young-Kyung; Yoshioka, Hifumi; Iwahashi, Yumiko

    2013-05-01

    The metabolic products resulting from the cultivation of F. asiaticum in agmatine were identified using capillary electrophoresis-time of flight mass spectrometry. Glyoxylic acid was detected from fungal cultures grown in agmatine, while it was absent in control cells. The abundance of other metabolic products of the glycolytic pathway also increased because of agmatine; however, there was no increase in the amounts of pyruvic acid or metabolites from the tricarboxylic acid cycle. Moreover, gene expression levels within Fusarium asiaticum exposed to agmatine were analyzed by DNA microarray. Changes in gene expression levels directed the changes in metabolic products. Our results suggest that acetyl coenzyme A, which is a starting substrate for the biosynthesis of deoxynivalenol (DON), was simultaneously produced by activated β-oxidation. Furthermore, the content of 4-aminobutyrate (GABA) was increased in the agmatine addition culture medium. GABA can be synthesized from agmatine through putrescine and might influence the regulation of DON-related genes.

  17. Metabolic-flux analysis of hydrogen production pathway in Citrobacter amalonaticus Y19

    Oh, You-Kwan; Kim, Mi-Sun [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-343 (Korea); Kim, Heung-Joo; Park, Sunghoon [Department of Chemical and Biochemical Engineering and Institute for Environmental Technology and Industry, Pusan National University, Busan 609-735 (Korea); Ryu, Dewey D.Y. [Biochemical Engineering Program, Department of Chemical Engineering and Material Science, University of California, Davis, CA 95616 (United States)

    2008-03-15

    For the newly isolated chemoheterotrophic bacterium Citrobacter amalonaticus Y19, anaerobic glucose metabolism and hydrogen (H{sub 2}) production pathway were studied using batch cultivation and an in silico metabolic-flux analysis. Batch cultivation was conducted under varying initial glucose concentration between 1.5 and 9.5 g/L with quantitative measurement of major metabolites to obtain accurate carbon material balance. The metabolic flux of Y19 was analyzed using a metabolic-pathway model which was constructed from 81 biochemical reactions. The linear optimization program MetaFluxNet was employed for the analysis. When the specific growth rate of cells was chosen as an objective function, the model described the batch culture characteristics of Ci. amalonaticus Y19 reasonably well. When the specific H{sub 2} production rate was selected as an objective function, on the other hand, the achievable maximal H{sub 2} production yield (8.7molH{sub 2}/mol glucose) and the metabolic pathway enabling the high H{sub 2} yield were identified. The pathway involved non-native NAD(P)-linked hydrogenase and H{sub 2} production from NAD(P)H which were supplied at a high rate from glucose degradation through the pentose phosphate pathway. (author)

  18. Metabolic engineering of yeast for fermentative production of flavonoids

    Rodriguez Prado, Edith Angelica; Strucko, Tomas; Stahlhut, Steen Gustav

    2017-01-01

    Yeast Saccharomyces cerevisiae was engineered for de novo production of six different flavonoids (naringenin, liquiritigenin, kaempferol, resokaempferol, quercetin, and fisetin) directly from glucose, without supplementation of expensive intermediates. This required reconstruction of long...... demonstrates the potential of flavonoid-producing yeast cell factories....

  19. Improvements in Fermentative Biological Hydrogen Production Through Metabolic Engineering

    Hallenbeck, P. C.; Ghosh, D.; Sabourin-Provost, G.

    2009-07-01

    Dramatically rising oil prices and increasing awareness of the dire environmental consequences of fossil fuel use, including startling effects of climate change, are refocusing attention world-wide on the search for alternative fuels. Hydrogen is poised to become an important future energy carrier. Renewable hydrogen production is pivotal in making it a truly sustainable replacement for fossil fuels. (Author)

  20. Improvements in Fermentative Biological Hydrogen Production Through Metabolic Engineering

    Hallenbeck, P. C.; Ghosh, D.; Sabourin-Provost, G.

    2009-01-01

    Dramatically rising oil prices and increasing awareness of the dire environmental consequences of fossil fuel use, including startling effects of climate change, are refocusing attention world-wide on the search for alternative fuels. Hydrogen is poised to become an important future energy carrier. Renewable hydrogen production is pivotal in making it a truly sustainable replacement for fossil fuels. (Author)

  1. Metabolic modeling of mixed substrate uptake for polyhydroxyalkanoate (PHA) production

    Jiang, Y.; Hebly, M.; Kleerebezem, R.; Muyzer, G.; van Loosdrecht, M.C.M.

    2011-01-01

    Polyhydroxyalkanoate (PHA) production by mixed microbial communities can be established in a two-stage process, consisting of a microbial enrichment step and a PHA accumulation step. In this study, a mathematical model was constructed for evaluating the influence of the carbon substrate composition

  2. Genome-scale metabolic analysis of Clostridium thermocellum for bioethanol production

    Brooks J Paul

    2010-03-01

    Full Text Available Abstract Background Microorganisms possess diverse metabolic capabilities that can potentially be leveraged for efficient production of biofuels. Clostridium thermocellum (ATCC 27405 is a thermophilic anaerobe that is both cellulolytic and ethanologenic, meaning that it can directly use the plant sugar, cellulose, and biochemically convert it to ethanol. A major challenge in using microorganisms for chemical production is the need to modify the organism to increase production efficiency. The process of properly engineering an organism is typically arduous. Results Here we present a genome-scale model of C. thermocellum metabolism, iSR432, for the purpose of establishing a computational tool to study the metabolic network of C. thermocellum and facilitate efforts to engineer C. thermocellum for biofuel production. The model consists of 577 reactions involving 525 intracellular metabolites, 432 genes, and a proteomic-based representation of a cellulosome. The process of constructing this metabolic model led to suggested annotation refinements for 27 genes and identification of areas of metabolism requiring further study. The accuracy of the iSR432 model was tested using experimental growth and by-product secretion data for growth on cellobiose and fructose. Analysis using this model captures the relationship between the reduction-oxidation state of the cell and ethanol secretion and allowed for prediction of gene deletions and environmental conditions that would increase ethanol production. Conclusions By incorporating genomic sequence data, network topology, and experimental measurements of enzyme activities and metabolite fluxes, we have generated a model that is reasonably accurate at predicting the cellular phenotype of C. thermocellum and establish a strong foundation for rational strain design. In addition, we are able to draw some important conclusions regarding the underlying metabolic mechanisms for observed behaviors of C. thermocellum

  3. Gravity control of growth form in brassica rapa and arabidopsis thaliana (brassicaceae): consequences for secondary metabolism

    Allen, J.; Bisbee, P.A.; Darnell, R.L.; Kuang, A.; Levine, L.H.; Musgrave, M.E.; van Loon, J.J.W.A.

    2009-01-01

    How gravity influences the growth form and flavor components of plants is of interest to the space program because plants could be used for food and life support during prolonged missions away from the planet, where that constant feature of Earth’s environment does not prevail. We used plant growth

  4. Dynamic 31phosphorus magnetic resonance spectroscopy of the quadriceps muscle: Metabolic changes resulting from two different forms of exercise

    Schunk, K.; Kersjes, W.; Schadmand-Fischer, S.; Thelen, M.

    1997-01-01

    Purpose: The aim of the present investigation was to examine the metabolism of the quadriceps muscles of normal young individuals using dynamic 31 phosphorus magnetic resonance spectroscopy. Methods: 22 normal individuals were examined in a 1.5 T-MRT using a 6 cm surface coil. The metabolic changes in the quadriceps muscle as shown by the phosphorus spectrum were evaluated during rest, exercise (isometric and isotonic exercise) and during a 36-second period of recovery. Results: The P i /PCr quotient rose from its resting value of 0.11±0.02 following exercise to a maximum of 0.83±0.47 (isometric) or 1.40±0.59 (isotonic) (difference p=0.0001). Half-time recovery of P i /PCr was 35±11 s or 31±10 s, respectively (p=0.13). During the recovery phase P i /PCr fell briefly but significantly below its rest value. Following an initial rise in pH, there was a continual fall. Minimum pH (6.68±0.21 and 6.53±0.27 respectively; p=0.01) occurred in the early recovery phase. The recovery process of pH values lasted longer following isotonic than after isometric exercise (half-value recovery time 229±72 s and 146±55 s, respectively; p=0.001). Conlcusion: Compared with isometric exercise, isotonic stress is more expensive in terms of metabolism. Dynamic 31 phosphorus MRT spectroscopy can differentiate changes in muscle metabolism during different forms of exercise. (orig.) [de

  5. Progress of succinic acid production from renewable resources: Metabolic and fermentative strategies.

    Jiang, Min; Ma, Jiangfeng; Wu, Mingke; Liu, Rongming; Liang, Liya; Xin, Fengxue; Zhang, Wenming; Jia, Honghua; Dong, Weiliang

    2017-12-01

    Succinic acid is a four-carbon dicarboxylic acid, which has attracted much interest due to its abroad usage as a precursor of many industrially important chemicals in the food, chemicals, and pharmaceutical industries. Facing the shortage of crude oil supply and demand of sustainable development, biological production of succinic acid from renewable resources has become a topic of worldwide interest. In recent decades, robust producing strain selection, metabolic engineering of model strains, and process optimization for succinic acid production have been developed. This review provides an overview of succinic acid producers and cultivation technology, highlight some of the successful metabolic engineering approaches. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production.

    Lu Jin

    Full Text Available Caffeine (1, 3, 7-trimethylxanthine and theobromine (3, 7-dimethylxanthine are the major purine alkaloids in plants, e.g., tea (Camellia sinensis and coffee (Coffea arabica. Caffeine is a major component of coffee and is used widely in food and beverage industries. Most of the enzymes involved in the caffeine biosynthetic pathway have been reported previously. Here, we demonstrated the biosynthesis of caffeine (0.38 mg/L by co-expression of Coffea arabica xanthosine methyltransferase (CaXMT and Camellia sinensis caffeine synthase (TCS in Saccharomyces cerevisiae. Furthermore, we endeavored to develop this production platform for making other purine-based alkaloids. To increase the catalytic activity of TCS in an effort to increase theobromine production, we identified four amino acid residues based on structural analyses of 3D-model of TCS. Two TCS1 mutants (Val317Met and Phe217Trp slightly increased in theobromine accumulation and simultaneously decreased in caffeine production. The application and further optimization of this biosynthetic platform are discussed.

  7. Metabolic engineering of Saccharomyces cerevisiae for caffeine and theobromine production.

    Jin, Lu; Bhuiya, Mohammad Wadud; Li, Mengmeng; Liu, XiangQi; Han, Jixiang; Deng, WeiWei; Wang, Min; Yu, Oliver; Zhang, Zhengzhu

    2014-01-01

    Caffeine (1, 3, 7-trimethylxanthine) and theobromine (3, 7-dimethylxanthine) are the major purine alkaloids in plants, e.g., tea (Camellia sinensis) and coffee (Coffea arabica). Caffeine is a major component of coffee and is used widely in food and beverage industries. Most of the enzymes involved in the caffeine biosynthetic pathway have been reported previously. Here, we demonstrated the biosynthesis of caffeine (0.38 mg/L) by co-expression of Coffea arabica xanthosine methyltransferase (CaXMT) and Camellia sinensis caffeine synthase (TCS) in Saccharomyces cerevisiae. Furthermore, we endeavored to develop this production platform for making other purine-based alkaloids. To increase the catalytic activity of TCS in an effort to increase theobromine production, we identified four amino acid residues based on structural analyses of 3D-model of TCS. Two TCS1 mutants (Val317Met and Phe217Trp) slightly increased in theobromine accumulation and simultaneously decreased in caffeine production. The application and further optimization of this biosynthetic platform are discussed.

  8. Mini-review: In vitro Metabolic Engineering for Biomanufacturing of High-value Products

    Weihua Guo

    Full Text Available With the breakthroughs in biomolecular engineering and synthetic biology, many valuable biologically active compound and commodity chemicals have been successfully manufactured using cell-based approaches in the past decade. However, because of the high complexity of cell metabolism, the identification and optimization of rate-limiting metabolic pathways for improving the product yield is often difficult, which represents a significant and unavoidable barrier of traditional in vivo metabolic engineering. Recently, some in vitro engineering approaches were proposed as alternative strategies to solve this problem. In brief, by reconstituting a biosynthetic pathway in a cell-free environment with the supplement of cofactors and substrates, the performance of each biosynthetic pathway could be evaluated and optimized systematically. Several value-added products, including chemicals, nutraceuticals, and drug precursors, have been biosynthesized as proof-of-concept demonstrations of in vitro metabolic engineering. This mini-review summarizes the recent progresses on the emerging topic of in vitro metabolic engineering and comments on the potential application of cell-free technology to speed up the “design-build-test” cycles of biomanufacturing. Keywords: Cell-free, Biosynthesis, Metabolic pathways, Design-build-test cycle

  9. Hydrogen peroxide production and myo-inositol metabolism as important traits for virulence of Mycoplasma hyopneumoniae.

    Ferrarini, M G; Mucha, S G; Parrot, D; Meiffren, G; Bachega, J F R; Comte, G; Zaha, A; Sagot, M F

    2018-04-06

    Mycoplasma hyopneumoniae is the causative agent of enzootic pneumonia. In our previous work, we reconstructed the metabolic models of this species along with two other mycoplasmas from the respiratory tract of swine: Mycoplasma hyorhinis, considered less pathogenic but which nonetheless causes disease and Mycoplasma flocculare, a commensal bacterium. We identified metabolic differences that partially explained their different levels of pathogenicity. One important trait was the production of hydrogen peroxide from the glycerol metabolism only in the pathogenic species. Another important feature was a pathway for the metabolism of myo-inositol in M. hyopneumoniae. Here, we tested these traits to understand their relation to the different levels of pathogenicity, comparing not only the species but also pathogenic and attenuated strains of M. hyopneumoniae. Regarding the myo-inositol metabolism, we show that only M. hyopneumoniae assimilated this carbohydrate and remained viable when myo-inositol was the primary energy source. Strikingly, only the two pathogenic strains of M. hyopneumoniae produced hydrogen peroxide in complex medium. We also show that this production was dependent on the presence of glycerol. Although further functional tests are needed, we present in this work two interesting metabolic traits of M. hyopneumoniae that might be directly related to its enhanced virulence. This article is protected by copyright. All rights reserved. © 2018 John Wiley & Sons Ltd.

  10. Leaching characteristics of the metal waste form from the electrometallurgical treatment process: Product consistency testing

    Johnson, S. G.; Keiser, D. D.; Frank, S. M.; DiSanto, T.; Noy, M.

    1999-01-01

    Argonne National Laboratory is developing an electrometallurgical treatment for spent fuel from the experimental breeder reactor II. A product of this treatment process is a metal waste form that incorporates the stainless steel cladding hulls, zirconium from the fuel and the fission products that are noble to the process, i.e., Tc, Ru, Nb, Pd, Rh, Ag. The nominal composition of this waste form is stainless steel/15 wt% zirconium/1--4 wt% noble metal fission products/1--2 wt % U. Leaching results are presented from several tests and sample types: (1) 2 week monolithic immersion tests on actual metal waste forms produced from irradiated cladding hulls, (2) long term (>2 years) pulsed flow tests on samples containing technetium and uranium and (3) crushed sample immersion tests on cold simulated metal waste form samples. The test results will be compared and their relevance for waste form product consistency testing discussed

  11. Mitochondrial biogenesis and energy production in differentiating murine stem cells: a functional metabolic study.

    Han, Sungwon; Auger, Christopher; Thomas, Sean C; Beites, Crestina L; Appanna, Vasu D

    2014-02-01

    The significance of metabolic networks in guiding the fate of the stem cell differentiation is only beginning to emerge. Oxidative metabolism has been suggested to play a major role during this process. Therefore, it is critical to understand the underlying mechanisms of metabolic alterations occurring in stem cells to manipulate the ultimate outcome of these pluripotent cells. Here, using P19 murine embryonal carcinoma cells as a model system, the role of mitochondrial biogenesis and the modulation of metabolic networks during dimethyl sulfoxide (DMSO)-induced differentiation are revealed. Blue native polyacrylamide gel electrophoresis (BN-PAGE) technology aided in profiling key enzymes, such as hexokinase (HK) [EC 2.7.1.1], glucose-6-phosphate isomerase (GPI) [EC 5.3.1.9], pyruvate kinase (PK) [EC 2.7.1.40], Complex I [EC 1.6.5.3], and Complex IV [EC 1.9.3.1], that are involved in the energy budget of the differentiated cells. Mitochondrial adenosine triphosphate (ATP) production was shown to be increased in DMSO-treated cells upon exposure to the tricarboxylic acid (TCA) cycle substrates, such as succinate and malate. The increased mitochondrial activity and biogenesis were further confirmed by immunofluorescence microscopy. Collectively, the results indicate that oxidative energy metabolism and mitochondrial biogenesis were sharply upregulated in DMSO-differentiated P19 cells. This functional metabolic and proteomic study provides further evidence that modulation of mitochondrial energy metabolism is a pivotal component of the cellular differentiation process and may dictate the final destiny of stem cells.

  12. Biobased production of alkanes and alkenes through metabolic engineering of microorganisms

    Kang, Min Kyoung; Nielsen, Jens

    2017-01-01

    Advancement in metabolic engineering of microorganisms has enabled bio-based production of a range of chemicals, and such engineered microorganism can be used for sustainable production leading to reduced carbon dioxide emission there. One area that has attained much interest is microbial...... hydrocarbon biosynthesis, and in particular, alkanes and alkenes are important high-value chemicals as they can be utilized for a broad range of industrial purposes as well as ‘drop-in’ biofuels. Some microorganisms have the ability to biosynthesize alkanes and alkenes naturally, but their production level...... is extremely low. Therefore, there have been various attempts to recruit other microbial cell factories for production of alkanes and alkenes by applying metabolic engineering strategies. Here we review different pathways and involved enzymes for alkane and alkene production and discuss bottlenecks...

  13. Metabolic engineering of microorganisms for biofuels production: from bugs to synthetic biology to fuels

    Kuk Lee, Sung; Chou, Howard; Ham, Timothy S.; Soon Lee, Taek; Keasling, Jay D.

    2009-12-02

    The ability to generate microorganisms that can produce biofuels similar to petroleum-based transportation fuels would allow the use of existing engines and infrastructure and would save an enormous amount of capital required for replacing the current infrastructure to accommodate biofuels that have properties significantly different from petroleum-based fuels. Several groups have demonstrated the feasibility of manipulating microbes to produce molecules similar to petroleum-derived products, albeit at relatively low productivity (e.g. maximum butanol production is around 20 g/L). For cost-effective production of biofuels, the fuel-producing hosts and pathways must be engineered and optimized. Advances in metabolic engineering and synthetic biology will provide new tools for metabolic engineers to better understand how to rewire the cell in order to create the desired phenotypes for the production of economically viable biofuels.

  14. Metabolic engineering of Corynebacterium glutamicum for fermentative production of chemicals in biorefinery.

    Baritugo, Kei-Anne; Kim, Hee Taek; David, Yokimiko; Choi, Jong-Il; Hong, Soon Ho; Jeong, Ki Jun; Choi, Jong Hyun; Joo, Jeong Chan; Park, Si Jae

    2018-05-01

    Bio-based production of industrially important chemicals provides an eco-friendly alternative to current petrochemical-based processes. Because of the limited supply of fossil fuel reserves, various technologies utilizing microbial host strains for the sustainable production of platform chemicals from renewable biomass have been developed. Corynebacterium glutamicum is a non-pathogenic industrial microbial species traditionally used for L-glutamate and L-lysine production. It is a promising species for industrial production of bio-based chemicals because of its flexible metabolism that allows the utilization of a broad spectrum of carbon sources and the production of various amino acids. Classical breeding, systems, synthetic biology, and metabolic engineering approaches have been used to improve its applications, ranging from traditional amino-acid production to modern biorefinery systems for production of value-added platform chemicals. This review describes recent advances in the development of genetic engineering tools and techniques for the establishment and optimization of metabolic pathways for bio-based production of major C2-C6 platform chemicals using recombinant C. glutamicum.

  15. Engineering the fatty acid metabolic pathway in Saccharomyces cerevisiae for advanced biofuel production

    Xiaoling Tang

    2015-12-01

    Full Text Available Fatty acid-derived fuels and chemicals have attracted a great deal of attention in recent decades, due to their following properties of high compatibility to gasoline-based fuels and existing infrastructure for their direct utilization, storage and distribution. The yeast Saccharomyces cerevisiae is the ideal biofuel producing candidate, based on the wealth of available genetic information and versatile tools designed to manipulate its metabolic pathways. Engineering the fatty acid metabolic pathways in S. cerevisiae is an effective strategy to increase its fatty acid biosynthesis and provide more pathway precursors for production of targeted products. This review summarizes the recent progress in metabolic engineering of yeast cells for fatty acids and fatty acid derivatives production, including the regulation of acetyl-CoA biosynthesis, NADPH production, fatty acid elongation, and the accumulation of activated precursors of fatty acids for converting enzymes. By introducing specific enzymes in the engineered strains, a powerful platform with a scalable, controllable and economic route for advanced biofuel production has been established. Keywords: Metabolic engineering, Fatty acid biosynthesis, Fatty acid derivatives, Saccharomyces cerevisiae

  16. Fumaric acid production in Saccharomyces cerevisiae by in silico aided metabolic engineering.

    Guoqiang Xu

    Full Text Available Fumaric acid (FA is a promising biomass-derived building-block chemical. Bio-based FA production from renewable feedstock is a promising and sustainable alternative to petroleum-based chemical synthesis. Here we report on FA production by direct fermentation using metabolically engineered Saccharomyces cerevisiae with the aid of in silico analysis of a genome-scale metabolic model. First, FUM1 was selected as the target gene on the basis of extensive literature mining. Flux balance analysis (FBA revealed that FUM1 deletion can lead to FA production and slightly lower growth of S. cerevisiae. The engineered S. cerevisiae strain obtained by deleting FUM1 can produce FA up to a concentration of 610±31 mg L(-1 without any apparent change in growth in fed-batch culture. FT-IR and (1H and (13C NMR spectra confirmed that FA was synthesized by the engineered S. cerevisiae strain. FBA identified pyruvate carboxylase as one of the factors limiting higher FA production. When the RoPYC gene was introduced, S. cerevisiae produced 1134±48 mg L(-1 FA. Furthermore, the final engineered S. cerevisiae strain was able to produce 1675±52 mg L(-1 FA in batch culture when the SFC1 gene encoding a succinate-fumarate transporter was introduced. These results demonstrate that the model shows great predictive capability for metabolic engineering. Moreover, FA production in S. cerevisiae can be efficiently developed with the aid of in silico metabolic engineering.

  17. Comparative study of hop-containing products on human cytochrome p450-mediated metabolism.

    Foster, Brian C; Kearns, Nikia; Arnason, John T; Saleem, Ammar; Ogrodowczyk, Carolina; Desjardins, Suzanne

    2009-06-10

    Thirty-five national and international brands of beer were examined for their potential to affect human cytochrome P450 (CYP)-mediated metabolism. They represented the two main categories of beer, ales and lagers, and included a number of specialty products including bitter (porter, stout), coffee, ice, wheat, Pilsner, and hemp seed. Aliquots were examined for nonvolatile soluble solids, effect on CYP metabolism and P-glycoprotein (Pgp) transport, and major alpha- and beta-hop acids. Wide variance was detected in contents of alcohol, nonvolatile suspended solids, and hop acids and in the potential to affect CYP-mediated metabolism and Pgp-mediated efflux transport. Many of the products affected CYP2C9-mediated metabolism, and only two (NRP 306 and 307) markedly affected CYP3A4; hence, some products have the capacity to affect drug safety. CYP3A4, CYP3A5, CYP3A7, and CYP19 (aromatase) inhibition to the log concentration of beta-acid content was significant with r(2) > 0.37, suggesting that these components can account for some of the variation in inhibition of CYP metabolism.

  18. Designing products as an integral part of choreography of interaction : the product's form as an integral part of movement

    Klooster, S.; Overbeeke, C.J.; Feijs, L.; Kyffin, S.; Young, B.

    2005-01-01

    Recent developments in design research concentrate on two themes (1) the unity of form, function and interaction and (2) the semantics of movement. The Design Movement approach incorporates unity of form, function and interaction through movement. Design Movement introduces the design of products as

  19. Role of glycolytic intermediate in regulation: Improving lycopene production in Escherichia coli by engineering metabolic control

    Farmer, W.R.; Liao, J.C.

    2001-06-01

    Metabolic engineering in the postgenomic era is expected to benefit from a full understanding of the biosynthetic capability of microorganisms as a result of the progress being made in bioinformatics and functional genomics. The immediate advantage of such information is to allow the rational design of novel pathways and the elimination of native reactions that are detrimental or unnecessary for the desired purpose. However, with the ability to manipulate metabolic pathways becoming more effective, metabolic engineering will need to face a new challenge: the reengineering of the regulatory hierarchy that controls gene expression in those pathways. In addition to constructing the genetic composition of a metabolic pathway, they propose that it will become just as important to consider the dynamics of pathways gene expression. It has been widely observed that high-level induction of a recombinant protein or pathway leads to growth retardation and reduced metabolic activity. These phenotypic characteristics result from the fact that the constant demands of production placed upon the cell interfere with its changing requirements for growth. They believe that this common situation in metabolic engineering can be alleviated by designing a dynamic controller that is able to sense the metabolic state of the cell and regulate the expression of the recombinant pathway accordingly. This approach, which is termed metabolic control engineering, involves redesigning the native regulatory circuits and applying them to the recombinant pathway. The general goal of such an effort will be to control the flux to the recombinant pathway adaptively according to the cell's metabolic state. The dynamically controlled recombinant pathway can potentially lead to enhanced production, minimized growth retardation, and reduced toxic by-product formation. The regulation of gene expression in response to the physiological state is also essential to the success of gene therapy. Here they

  20. Metabolic engineering of Clostridium autoethanogenum for selective alcohol production.

    Liew, Fungmin; Henstra, Anne M; Kӧpke, Michael; Winzer, Klaus; Simpson, Sean D; Minton, Nigel P

    2017-03-01

    Gas fermentation using acetogenic bacteria such as Clostridium autoethanogenum offers an attractive route for production of fuel ethanol from industrial waste gases. Acetate reduction to acetaldehyde and further to ethanol via an aldehyde: ferredoxin oxidoreductase (AOR) and alcohol dehydrogenase has been postulated alongside the classic pathway of ethanol formation via a bi-functional aldehyde/alcohol dehydrogenase (AdhE). Here we demonstrate that AOR is critical to ethanol formation in acetogens and inactivation of AdhE led to consistently enhanced autotrophic ethanol production (up to 180%). Using ClosTron and allelic exchange mutagenesis, which was demonstrated for the first time in an acetogen, we generated single mutants as well as double mutants for both aor and adhE isoforms to confirm the role of each gene. The aor1+2 double knockout strain lost the ability to convert exogenous acetate, propionate and butyrate into the corresponding alcohols, further highlighting the role of these enzymes in catalyzing the thermodynamically unfavourable reduction of carboxylic acids into alcohols. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Genome Sequencing of Streptomyces atratus SCSIOZH16 and Activation Production of Nocardamine via Metabolic Engineering

    Yan Li

    2018-06-01

    Full Text Available The Actinomycetes are metabolically flexible microorganisms capable of producing a wide range of interesting compounds, including but by no means limited to, siderophores which have high affinity for ferric iron. In this study, we report the complete genome sequence of marine-derived Streptomyces atratus ZH16 and the activation of an embedded siderophore gene cluster via the application of metabolic engineering methods. The S. atratus ZH16 genome reveals that this strain has the potential to produce 26 categories of natural products (NPs barring the ilamycins. Our activation studies revealed S. atratus SCSIO ZH16 to be a promising source of the production of nocardamine-type (desferrioxamine compounds which are important in treating acute iron intoxication and performing ecological remediation. We conclude that metabolic engineering provides a highly effective strategy by which to discover drug-like compounds and new NPs in the genomic era.

  2. Metabolic Engineering of Oleaginous Yeasts for Production of Fuels and Chemicals

    Shuobo Shi

    2017-11-01

    Full Text Available Oleaginous yeasts have been increasingly explored for production of chemicals and fuels via metabolic engineering. Particularly, there is a growing interest in using oleaginous yeasts for the synthesis of lipid-related products due to their high lipogenesis capability, robustness, and ability to utilize a variety of substrates. Most of the metabolic engineering studies in oleaginous yeasts focused on Yarrowia that already has plenty of genetic engineering tools. However, recent advances in systems biology and synthetic biology have provided new strategies and tools to engineer those oleaginous yeasts that have naturally high lipid accumulation but lack genetic tools, such as Rhodosporidium, Trichosporon, and Lipomyces. This review highlights recent accomplishments in metabolic engineering of oleaginous yeasts and recent advances in the development of genetic engineering tools in oleaginous yeasts within the last 3 years.

  3. Teratogenicity and metabolism of water-soluble forms of vitamin A in the pregnant rat

    Gunning, D.B.; Barua, A.B.; Olson, J.A.

    1990-01-01

    Retinoyl β-glucuronide, unlike retinoic acid, has been shown to be non-teratogenic when administered orally, even in large doses, to pregnant rats. The degree to which water-solubility is associated with low teratogenicity is not known. Other water-soluble forms of vitamin A have now been synthesized in our laboratory and are being evaluated for teratogenicity. New water-soluble forms of vitamin A were administered orally to pregnant Sprague-Dawley rats in a single dose of 0.35 mmole/kg bw on day 8 of gestation. On day 19, the dams were sacrificed and the litters were examined. Control animals received either vehicle only or an equivalent dose of all-trans retinoic acid. Maternal and fetal tissues were taken and analyzed by HPLC for vitamin A metabolites. In another experiment, a large single oral dose of the radiolabelled water-soluble compound was administered on day 10. At either 30 minutes or 1 hour after the dose, dams were sacrificed and the embryos analyzed both for radioactivity and for specific metabolites. In contrast to retinoyl β-glucuronide, retinoyl β-glucose is highly teratogenic under identical conditions. Thus, water-solubility does not seem to be the determining factor in the teratogenicity of retinoic acid conjugates

  4. Impact of synthetic biology and metabolic engineering on industrial production of fine chemicals.

    Jullesson, David; David, Florian; Pfleger, Brian; Nielsen, Jens

    2015-11-15

    Industrial bio-processes for fine chemical production are increasingly relying on cell factories developed through metabolic engineering and synthetic biology. The use of high throughput techniques and automation for the design of cell factories, and especially platform strains, has played an important role in the transition from laboratory research to industrial production. Model organisms such as Saccharomyces cerevisiae and Escherichia coli remain widely used host strains for industrial production due to their robust and desirable traits. This review describes some of the bio-based fine chemicals that have reached the market, key metabolic engineering tools that have allowed this to happen and some of the companies that are currently utilizing these technologies for developing industrial production processes. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Impact of synthetic biology and metabolic engineering on industrial production of fine chemicals

    Jullesson, David; David, Florian; Pfleger, Brian

    2015-01-01

    Industrial bio-processes for fine chemical production are increasingly relying on cell factories developed through metabolic engineering and synthetic biology. The use of high throughput techniques and automation for the design of cell factories, and especially platform strains, has played...... chemicals that have reached the market, key metabolic engineering tools that have allowed this to happen and some of the companies that are currently utilizing these technologies for developing industrial production processes....... an important role in the transition from laboratory research to industrial production. Model organisms such as Saccharomyces cerevisiae and Escherichia coli remain widely used host strains for industrial production due to their robust and desirable traits. This review describes some of the bio-based fine...

  6. Production of anthocyanins in metabolically engineered microorganisms: Current status and perspectives

    Jian Zha

    2017-12-01

    Full Text Available Microbial production of plant-derived natural products by engineered microorganisms has achieved great success thanks to large extend to metabolic engineering and synthetic biology. Anthocyanins, the water-soluble colored pigments found in terrestrial plants that are responsible for the red, blue and purple coloration of many flowers and fruits, are extensively used in food and cosmetics industry; however, their current supply heavily relies on complex extraction from plant-based materials. A promising alternative is their sustainable production in metabolically engineered microbes. Here, we review the recent progress on anthocyanin biosynthesis in engineered bacteria, with a special focus on the systematic engineering modifications such as selection and engineering of biosynthetic enzymes, engineering of transportation, regulation of UDP-glucose supply, as well as process optimization. These promising engineering strategies will facilitate successful microbial production of anthocyanins in industry in the near future.

  7. Improving production of ?-lactam antibiotics by Penicillium chrysogenum : Metabolic engineering based on transcriptome analysis

    Veiga, T.

    2012-01-01

    In Chapters 2-5 of this thesis, the applicability of transcriptome analysis to guide metabolic engineering strategies in P. chrysogenum is explored by investigating four cellular processes that are of potential relevance for industrial production of ?-lactam antibiotics: - Regulation of secondary

  8. Production of xylitol by a Coniochaeta ligniaria strain tolerant of inhibitors and defective in xylose metabolism

    In conversion of biomass to fuels or chemicals, inhibitory compounds arising from physical-chemical pretreatment of the feedstock can interfere with fermentation of the sugars to product. Fungal strain Coniochaeta ligniaria NRRL30616, metabolizes the furan aldehydes furfural and 5-hydroxymethylfurfu...

  9. Nitrification-driven forms of nitrogen metabolism in microbial mat communities thriving along an ammonium-enriched subsurface geothermal stream

    Nishizawa, Manabu; Koba, Keisuke; Makabe, Akiko; Yoshida, Naohiro; Kaneko, Masanori; Hirao, Shingo; Ishibashi, Jun-ichiro; Yamanaka, Toshiro; Shibuya, Takazo; Kikuchi, Tohru; Hirai, Miho; Miyazaki, Junichi; Nunoura, Takuro; Takai, Ken

    2013-07-01

    We report here the concurrence and interaction among forms of nitrogen metabolism in thermophilic microbial mat communities that developed in an ammonium-abundant subsurface geothermal stream. First, the physical and chemical conditions of the stream water at several representative microbial mat habitats (including upper, middle and downstream sites) were characterized. A thermodynamic calculation using these physical and chemical conditions predicted that nitrification consisting of ammonia and nitrite oxidations would provide one of the largest energy yields of chemolithotrophic metabolisms. Second, near-complete prokaryotic 16S rRNA gene clone analysis was conducted for representative microbial mat communities at the upper, middle and downstream sites. The results indicated a dynamic shift in the 16S rRNA gene phylotype composition through physical and chemical variations of the stream water. The predominant prokaryotic components varied from phylotypes related to hydrogeno (H2)- and thio (S)-trophic Aquificales, thermophilic methanotrophs and putative ammonia-oxidizing Archaea (AOA) located upstream (72 °C) to the phylotypes affiliated with putative AOA and nitrite-oxidizing bacteria (NOB) located at the middle and downstream sites (65 and 57 °C, respectively). In addition, the potential in situ metabolic activities of different forms of nitrogen metabolism were estimated through laboratory experiments using bulk microbial mat communities. Finally, the compositional and isotopic variation in nitrogen compounds was investigated in the stream water flowing over the microbial mats and in the interstitial water inside the mats. Although the stream water was characterized by a gradual decrease in the total ammonia concentration (ΣNH3: the sum of ammonia and ammonium concentrations) and a gradual increase in the total concentration of nitrite and nitrate (NO2- + NO3-), the total inorganic nitrogen concentration (TIN: the sum of ΣNH3, NO2- and NO3- concentrations

  10. Metabolic engineering for the microbial production of isoprenoids: Carotenoids and isoprenoid-based biofuels

    Fu-Xing Niu

    2017-09-01

    Full Text Available Isoprenoids are the most abundant and highly diverse group of natural products. Many isoprenoids have been used for pharmaceuticals, nutraceuticals, flavors, cosmetics, food additives and biofuels. Carotenoids and isoprenoid-based biofuels are two classes of important isoprenoids. These isoprenoids have been produced microbially through metabolic engineering and synthetic biology efforts. Herein, we briefly review the engineered biosynthetic pathways in well-characterized microbial systems for the production of carotenoids and several isoprenoid-based biofuels.

  11. Biobased production of alkanes and alkenes through metabolic engineering of microorganisms

    Kang, Min Kyoung; Nielsen, Jens

    2017-01-01

    Advancement in metabolic engineering of microorganisms has enabled bio-based production of a range of chemicals, and such engineered microorganism can be used for sustainable production leading to reduced carbon dioxide emission there. One area that has attained much interest is microbial hydrocarbon biosynthesis, and in particular, alkanes and alkenes are important high-value chemicals as they can be utilized for a broad range of industrial purposes as well as ?drop-in? biofuels. Some microo...

  12. Ketocarotenoid Production in Soybean Seeds through Metabolic Engineering.

    Emily C Pierce

    Full Text Available The pink or red ketocarotenoids, canthaxanthin and astaxanthin, are used as feed additives in the poultry and aquaculture industries as a source of egg yolk and flesh pigmentation, as farmed animals do not have access to the carotenoid sources of their wild counterparts. Because soybean is already an important component in animal feed, production of these carotenoids in soybean could be a cost-effective means of delivery. In order to characterize the ability of soybean seed to produce carotenoids, soybean cv. Jack was transformed with the crtB gene from Pantoea ananatis, which codes for phytoene synthase, an enzyme which catalyzes the first committed step in the carotenoid pathway. The crtB gene was engineered together in combinations with ketolase genes (crtW from Brevundimonas sp. strain SD212 and bkt1 from Haematococcus pluvialis to produce ketocarotenoids; all genes were placed under the control of seed-specific promoters. HPLC results showed that canthaxanthin is present in the transgenic seeds at levels up to 52 μg/g dry weight. Transgenic seeds also accumulated other compounds in the carotenoid pathway, such as astaxanthin, lutein, β-carotene, phytoene, α-carotene, lycopene, and β-cryptoxanthin, whereas lutein was the only one of these detected in non-transgenic seeds. The accumulation of astaxanthin, which requires a β-carotene hydroxylase in addition to a β-carotene ketolase, in the transgenic seeds suggests that an endogenous soybean enzyme is able to work in combination with the ketolase transgene. Soybean seeds that accumulate ketocarotenoids could potentially be used in animal feed to reduce or eliminate the need for the costly addition of these compounds.

  13. Worldwide research productivity in the field of endocrinology and metabolism--a bibliometric analysis.

    Zhao, Xiyan; Ye, Ru; Zhao, Linhua; Lin, Yiqun; Huang, Wenjing; He, Xinhui; Lian, Fengmei; Tong, Xiaolin

    2015-01-01

    Recently, significant contributions to the study of endocrinology and metabolism have been made. The national contribution, however, has not been reported. The aim of this study was to assess national efforts in the field of endocrinology and metabolism. A Web of Science search was performed using subject categories "endocrinology & metabolism" to identify articles published from 2010 to 2014. The total and per capita numbers of articles and citations were analysed for different countries. A total of 79,394 articles were published on endocrinology and metabolism from 2010 to 2014. Most were published in North America, East Asia, and Europe. The majority (82.28%) were reported by authors in high-income countries, 17.64% were published in middle-income countries, and only 0.08% were published in low-income countries. Authors in the United States published the most articles (27.38%), followed by China (7.22%), Italy (5.70%), the United Kingdom (5.6%), and Japan (5.54%). Articles published by authors in the United States had the most citations (260,934). A positive correlation was found between the number of publications and population/gross domestic product (GDP; p endocrinology and metabolism articles were published by authors from high-income countries with few from low-income countries. The United States was the most productive country. However, when population size and GDP were considered, some European countries were ranked higher.

  14. In vivo effects of Faizol Ubat Batuk, a herbal product on aminopyrine metabolism in rat hepatocytes

    Abas Hj Hussin

    2011-09-01

    Full Text Available Traditional medicines, in particular herbal products, have been used abundantly over the years in curing several diseases. Pharmacological interactions of herbal products with modern drugs, however, remain to some extent unknown. Herein, we examined whether co-administration of Faizol Ubat Batuk (FUB, a mixture of aqueous extract of different plants, modifies the metabolism of aminopyrine, a conventional analgesic drug, in rat liver. We used rat hepatocytes outfitted by collagenase perfusion technique. Determination of aminopyrine n-demethylase activity was performed using the Nash colorimetric method, by measuring the amount of formaldehyde produced. Compared to control treatment, FUB significantly increased the hepatic metabolism of aminopyrine in healthy adult male rats. In contrast, the hepatic metabolism of aminopyrine in adult female rats was decreased. Besides, a biphasic effect in n-demethylase activity was observed in young male rats treated with FUB. In a subsequent experiment, FUB did not change the metabolism of aminopyrine in streptozotocin (STZ-diabetic adult male rats. In conclusion, administration of FUB could affect phase I aminopyrine metabolism in rat heptocytes. In addition, the effects of FUB on hepatic n-demethylase activity were gender and disease dependent.

  15. Rewiring Host Lipid Metabolism by Large Viruses Determines the Fate of Emiliania huxleyi, a Bloom-Forming Alga in the Ocean[C][W][OPEN

    Rosenwasser, Shilo; Mausz, Michaela A.; Schatz, Daniella; Sheyn, Uri; Malitsky, Sergey; Aharoni, Asaph; Weinstock, Eyal; Tzfadia, Oren; Ben-Dor, Shifra; Feldmesser, Ester; Pohnert, Georg; Vardi, Assaf

    2014-01-01

    Marine viruses are major ecological and evolutionary drivers of microbial food webs regulating the fate of carbon in the ocean. We combined transcriptomic and metabolomic analyses to explore the cellular pathways mediating the interaction between the bloom-forming coccolithophore Emiliania huxleyi and its specific coccolithoviruses (E. huxleyi virus [EhV]). We show that EhV induces profound transcriptome remodeling targeted toward fatty acid synthesis to support viral assembly. A metabolic shift toward production of viral-derived sphingolipids was detected during infection and coincided with downregulation of host de novo sphingolipid genes and induction of the viral-encoded homologous pathway. The depletion of host-specific sterols during lytic infection and their detection in purified virions revealed their novel role in viral life cycle. We identify an essential function of the mevalonate-isoprenoid branch of sterol biosynthesis during infection and propose its downregulation as an antiviral mechanism. We demonstrate how viral replication depends on the hijacking of host lipid metabolism during the chemical “arms race” in the ocean. PMID:24920329

  16. Effect of aluminum on metabolism of organic acids and chemical forms of aluminum in root tips of Eucalyptus camaldulensis Dehnh.

    Ikka, Takashi; Ogawa, Tsuyoshi; Li, Donghua; Hiradate, Syuntaro; Morita, Akio

    2013-10-01

    Eucalyptus (Eucalyptus camaldulensis) has relatively high resistance to aluminum (Al) toxicity than the various herbaceous plants and model plant species. To investigate Al-tolerance mechanism, the metabolism of organic acids and the chemical forms of Al in the target site (root tips) in Eucalyptus was investigated. To do this, 2-year old rooted cuttings of E. camaldulensis were cultivated in half-strength Hoagland solution (pH 4.0) containing Al (0, 0.25, 0.5, 1.0, 2.5 and 5.0mM) salts for 5weeks; growth was not affected at concentrations up to 2.5mM even with Al concentration reaching 6000μgg(-1) DW. In roots, the citrate content also increased with increasing Al application. Concurrently, the activities of aconitase and NADP(+)-isocitrate dehydrogenase, which catalyze the decomposition of citrate, decreased. On the other hand, the activity of citrate synthase was not affected at concentrations up to 2.5mM Al. (27)Al-NMR spectroscopic analyses were carried out where it was found that Al-citrate complexes were a major chemical form present in cell sap of root tips. These findings suggested that E. camaldulensis detoxifies Al by forming Al-citrate complexes, and that this is achieved through Al-induced citrate accumulation in root tips via suppression of the citrate decomposition pathway. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Towards Mass Production by High Performance Transfer Press in Micro Bulk Forming

    Mahshid, Rasoul; Hansen, Hans Nørgaard; Arentoft, Mogens

    2014-01-01

    cold bulk forming. A methodology for selection of linear motors on the bases of the process parameters was obtained. In order to examine the effectiveness of the machine, specific geometry was investigated for production. Kinematic parameters were found for a production rate of 200 strokes per minute...

  18. A modular metabolic engineering approach for the production of 1,2-propanediol from glycerol by Saccharomyces cerevisiae.

    Islam, Zia-Ul; Klein, Mathias; Aßkamp, Maximilian R; Ødum, Anders S R; Nevoigt, Elke

    2017-11-01

    Compared to sugars, a major advantage of using glycerol as a feedstock for industrial bioprocesses is the fact that this molecule is more reduced than sugars. A compound whose biotechnological production might greatly profit from the substrate's higher reducing power is 1,2-propanediol (1,2-PDO). Here we present a novel metabolic engineering approach to produce 1,2-PDO from glycerol in S. cerevisiae. Apart from implementing the heterologous methylglyoxal (MG) pathway for 1,2-PDO formation from dihydroxyacetone phosphate (DHAP) and expressing a heterologous glycerol facilitator, the employed genetic modifications included the replacement of the native FAD-dependent glycerol catabolic pathway by the 'DHA pathway' for delivery of cytosolic NADH and the reduction of triosephosphate isomerase (TPI) activity for increased precursor (DHAP) supply. The choice of the medium had a crucial impact on both the strength of the metabolic switch towards fermentation in general (as indicated by the production of ethanol and 1,2-PDO) and on the ratio at which these two fermentation products were formed. For example, virtually no 1,2-PDO but only ethanol was formed in synthetic glycerol medium with urea as the nitrogen source. When nutrient-limited complex YG medium was used, significant amounts of 1,2-PDO were formed and it became obvious that the concerted supply of NADH and DHAP are essential for boosting 1,2-PDO production. Additionally, optimizing the flux into the MG pathway improved 1,2-PDO formation at the expense of ethanol. Cultivation of the best-performing strain in YG medium and a controlled bioreactor set-up resulted in a maximum titer of > 4gL -1 1,2-PDO which, to the best of our knowledge, has been the highest titer of 1,2-PDO obtained in yeast so far. Surprisingly, significant 1,2-PDO production was also obtained in synthetic glycerol medium after changing the nitrogen source towards ammonium sulfate and adding a buffer. Copyright © 2017 International Metabolic

  19. Metabolic engineering in chemolithoautotrophic hosts for the production of fuels and chemicals.

    Nybo, S Eric; Khan, Nymul E; Woolston, Benjamin M; Curtis, Wayne R

    2015-07-01

    The ability of autotrophic organisms to fix CO2 presents an opportunity to utilize this 'greenhouse gas' as an inexpensive substrate for biochemical production. Unlike conventional heterotrophic microorganisms that consume carbohydrates and amino acids, prokaryotic chemolithoautotrophs have evolved the capacity to utilize reduced chemical compounds to fix CO2 and drive metabolic processes. The use of chemolithoautotrophic hosts as production platforms has been renewed by the prospect of metabolically engineered commodity chemicals and fuels. Efforts such as the ARPA-E electrofuels program highlight both the potential and obstacles that chemolithoautotrophic biosynthetic platforms provide. This review surveys the numerous advances that have been made in chemolithoautotrophic metabolic engineering with a focus on hydrogen oxidizing bacteria such as the model chemolithoautotrophic organism (Ralstonia), the purple photosynthetic bacteria (Rhodobacter), and anaerobic acetogens. Two alternative strategies of microbial chassis development are considered: (1) introducing or enhancing autotrophic capabilities (carbon fixation, hydrogen utilization) in model heterotrophic organisms, or (2) improving tools for pathway engineering (transformation methods, promoters, vectors etc.) in native autotrophic organisms. Unique characteristics of autotrophic growth as they relate to bioreactor design and process development are also discussed in the context of challenges and opportunities for genetic manipulation of organisms as production platforms. Copyright © 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  20. Yeast cell metabolism investigated by CO{_2} production and soft X-ray irradiation

    Masini, A.; Batani, D.; Previdi, F.; Milani, M.; Pozzi, A.; Turcu, E.; Huntington, S.; Takeyasu, H.

    1999-01-01

    Results obtained using a new technique for studying cell metabolism are presented. The technique, consisting in CO2 production monitoring, has been applied to Saccharomyces cerevisiae yeast cells. Also the cells were irradiated using the soft X-ray laser-plasma source at Rutherford Appleton Laboratory with the aim of producing a damage of metabolic processes at the wall level, responsible for fermentation, without great interference with respiration, taking place in mitochondria, and DNA activity. The source was calibrated with PIN diodes and X-ray spectrometers and used Teflon stripes as target, emitting X-rays at about 0.9 keV, with a very low penetration in biological material. X-ray doses delivered to the different cell compartments were calculated following a Lambert-Bouguet-Beer law. Immediately after irradiation, the damage to metabolic activity was measured again by monitoring CO2 production. Results showed a general reduction in gas production by irradiated samples, together with non-linear and non-monotone response to dose. There was also evidence of oscillations in cell metabolic activity and of X-ray induced changes in oscillation frequency.

  1. Metabolic flux profiling of MDCK cells during growth and canine adenovirus vector production.

    Carinhas, Nuno; Pais, Daniel A M; Koshkin, Alexey; Fernandes, Paulo; Coroadinha, Ana S; Carrondo, Manuel J T; Alves, Paula M; Teixeira, Ana P

    2016-03-23

    Canine adenovirus vector type 2 (CAV2) represents an alternative to human adenovirus vectors for certain gene therapy applications, particularly neurodegenerative diseases. However, more efficient production processes, assisted by a greater understanding of the effect of infection on producer cells, are required. Combining [1,2-(13)C]glucose and [U-(13)C]glutamine, we apply for the first time (13)C-Metabolic flux analysis ((13)C-MFA) to study E1-transformed Madin-Darby Canine Kidney (MDCK) cells metabolism during growth and CAV2 production. MDCK cells displayed a marked glycolytic and ammoniagenic metabolism, and (13)C data revealed a large fraction of glutamine-derived labelling in TCA cycle intermediates, emphasizing the role of glutamine anaplerosis. (13)C-MFA demonstrated the importance of pyruvate cycling in balancing glycolytic and TCA cycle activities, as well as occurrence of reductive alphaketoglutarate (AKG) carboxylation. By turn, CAV2 infection significantly upregulated fluxes through most central metabolism, including glycolysis, pentose-phosphate pathway, glutamine anaplerosis and, more prominently, reductive AKG carboxylation and cytosolic acetyl-coenzyme A formation, suggestive of increased lipogenesis. Based on these results, we suggest culture supplementation strategies to stimulate nucleic acid and lipid biosynthesis for improved canine adenoviral vector production.

  2. Metabolic engineering of mannitol production in Lactococcus lactis: influence of overexpression of mannitol 1-phosphate dehydrogenase in different genetic backgrounds.

    Wisselink, H Wouter; Mars, Astrid E; van der Meer, Pieter; Eggink, Gerrit; Hugenholtz, Jeroen

    2004-07-01

    To obtain a mannitol-producing Lactococcus lactis strain, the mannitol 1-phosphate dehydrogenase gene (mtlD) from Lactobacillus plantarum was overexpressed in a wild-type strain, a lactate dehydrogenase(LDH)-deficient strain, and a strain with reduced phosphofructokinase activity. High-performance liquid chromatography and (13)C nuclear magnetic resonance analysis revealed that small amounts (<1%) of mannitol were formed by growing cells of mtlD-overexpressing LDH-deficient and phosphofructokinase-reduced strains, whereas resting cells of the LDH-deficient transformant converted 25% of glucose into mannitol. Moreover, the formed mannitol was not reutilized upon glucose depletion. Of the metabolic-engineering strategies investigated in this work, mtlD-overexpressing LDH-deficient L. lactis seemed to be the most promising strain for mannitol production.

  3. Seasonal Oxygen Dynamics in a Warm Temperate Estuary: Effects of Hydrologic Variability on Measurements of Primary Production, Respiration, and Net Metabolism

    Seasonal responses in estuarine metabolism (primary production, respiration, and net metabolism) were examined using two complementary approaches. Total ecosystem metabolism rates were calculated from dissolved oxygen time series using Odum’s open water method. Water column rates...

  4. Metabolic Engineering of the Shikimate Pathway for Production of Aromatics and Derived Compounds—Present and Future Strain Construction Strategies

    Nils J. H. Averesch

    2018-03-01

    Full Text Available The aromatic nature of shikimate pathway intermediates gives rise to a wealth of potential bio-replacements for commonly fossil fuel-derived aromatics, as well as naturally produced secondary metabolites. Through metabolic engineering, the abundance of certain intermediates may be increased, while draining flux from other branches off the pathway. Often targets for genetic engineering lie beyond the shikimate pathway, altering flux deep in central metabolism. This has been extensively used to develop microbial production systems for a variety of compounds valuable in chemical industry, including aromatic and non-aromatic acids like muconic acid, para-hydroxybenzoic acid, and para-coumaric acid, as well as aminobenzoic acids and aromatic α-amino acids. Further, many natural products and secondary metabolites that are valuable in food- and pharma-industry are formed outgoing from shikimate pathway intermediates. (Reconstruction of such routes has been shown by de novo production of resveratrol, reticuline, opioids, and vanillin. In this review, strain construction strategies are compared across organisms and put into perspective with requirements by industry for commercial viability. Focus is put on enhancing flux to and through shikimate pathway, and engineering strategies are assessed in order to provide a guideline for future optimizations.

  5. Global metabolic rewiring for improved CO2 fixation and chemical production in cyanobacteria

    Kanno, Masahiro; Carroll, Austin L.; Atsumi, Shota

    2017-03-01

    Cyanobacteria have attracted much attention as hosts to recycle CO2 into valuable chemicals. Although cyanobacteria have been engineered to produce various compounds, production efficiencies are too low for commercialization. Here we engineer the carbon metabolism of Synechococcus elongatus PCC 7942 to improve glucose utilization, enhance CO2 fixation and increase chemical production. We introduce modifications in glycolytic pathways and the Calvin Benson cycle to increase carbon flux and redirect it towards carbon fixation. The engineered strain efficiently uses both CO2 and glucose, and produces 12.6 g l-1 of 2,3-butanediol with a rate of 1.1 g l-1 d-1 under continuous light conditions. Removal of native regulation enables carbon fixation and 2,3-butanediol production in the absence of light. This represents a significant step towards industrial viability and an excellent example of carbon metabolism plasticity.

  6. Study on substrate metabolism process of saline waste sludge and its biological hydrogen production potential.

    Zhang, Zengshuai; Guo, Liang; Li, Qianqian; Zhao, Yangguo; Gao, Mengchun; She, Zonglian

    2017-07-01

    With the increasing of high saline waste sludge production, the treatment and utilization of saline waste sludge attracted more and more attention. In this study, the biological hydrogen production from saline waste sludge after heating pretreatment was studied. The substrate metabolism process at different salinity condition was analyzed by the changes of soluble chemical oxygen demand (SCOD), carbohydrate and protein in extracellular polymeric substances (EPS), and dissolved organic matters (DOM). The excitation-emission matrix (EEM) with fluorescence regional integration (FRI) was also used to investigate the effect of salinity on EPS and DOM composition during hydrogen fermentation. The highest hydrogen yield of 23.6 mL H 2 /g VSS and hydrogen content of 77.6% were obtained at 0.0% salinity condition. The salinity could influence the hydrogen production and substrate metabolism of waste sludge.

  7. Engineering yeast metabolism for production of terpenoids for use as perfume ingredients, pharmaceuticals and biofuels.

    Zhang, Yueping; Nielsen, Jens; Liu, Zihe

    2017-12-01

    Terpenoids represent a large class of natural products with significant commercial applications. These chemicals are currently mainly obtained through extraction from plants and microbes or through chemical synthesis. However, these sources often face challenges of unsustainability and low productivity. In order to address these issues, Escherichia coli and yeast have been metabolic engineered to produce non-native terpenoids. With recent reports of engineering yeast metabolism to produce several terpenoids at high yields, it has become possible to establish commercial yeast production of terpenoids that find applications as perfume ingredients, pharmaceuticals and advanced biofuels. In this review, we describe the strategies to rewire the yeast pathway for terpenoid biosynthesis. Recent advances will be discussed together with challenges and perspectives of yeast as a cell factory to produce different terpenoids. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Shikimic acid production in Escherichia coli: From classical metabolic engineering strategies to omics applied to improve its production

    Juan Andrés Martínez

    2015-09-01

    Full Text Available Shikimic acid (SA is an intermediate of the SA pathway that is present in bacteria and plants. SA has gained great interest because it is a precursor in the synthesis of the drug oseltamivir phosphate (OSF, an efficient inhibitor of the neuraminidase enzyme of diverse seasonal influenza viruses, the avian influenza virus H5N1, and the human influenza virus H1N1. For the purposes of OSF production, SA is extracted from the pods of Chinese star anise plants (Illicium spp., yielding up to 17% of SA (dry basis content. The high demand for OSF necessary to manage a major influenza outbreak is not adequately met by industrial production using SA from plants sources. As the SA pathway is present in the model bacteria Escherichia coli, several intuitive metabolically engineered strains have been applied for its successful overproduction by biotechnological processes, resulting in strains producing up to 71 g/L of SA, with high conversion yields of up to 0.42 (mol SA/mol Glc, in both batch and fed-batch cultures using complex fermentation broths, including glucose as a carbon source and yeast extract. Global transcriptomic analyses have been performed in SA producing strains, resulting in the identification of possible key target genes for the design of a rational strain improvement strategy. Because possible target genes are involved in the transport, catabolism and interconversion of different carbon sources and metabolic intermediates outside the central carbon metabolism and SA pathways, as genes involved in diverse cellular stress responses, the development of rational cellular strain improvement strategies based on omics data constitutes a challenging task to improve SA production in currently overproducing engineered strains. In this review, we discuss the main metabolic engineering strategies that have been applied for the development of efficient SA producing strains, as the perspective of omics analysis has focused on further strain improvement

  9. Improvements in or relating to the production of metal-containing material in particulate form

    Woodhead, J.L.; Scott, K.T.B.; Ball, P.W.

    1977-01-01

    The process described refers mainly to production of the material in the form of very small spheres. It comprises forming a metal compound-containing gel precipitate by mixing a solution or sol of the metal compound with a soluble organic polymer and contacting the mixture with a precipitating reagent to precipitate the metal as an insoluble compound bound with the polymer. The precipitate is then subjected in the liquid phase to a breaking down and dispersing process to produce an intermediate product suitable for spray drying, and the intermediate product is spray dried to form the particulate product. The breaking down and dispersing process may be performed by means of a colloid mill or vibratory stirrer. Examples of application of the process are described. (U.K.)

  10. Dropwise additive manufacturing of pharmaceutical products for solvent-based dosage forms.

    Hirshfield, Laura; Giridhar, Arun; Taylor, Lynne S; Harris, Michael T; Reklaitis, Gintaras V

    2014-02-01

    In recent years, the US Food and Drug Administration has encouraged pharmaceutical companies to develop more innovative and efficient manufacturing methods with improved online monitoring and control. Mini-manufacturing of medicine is one such method enabling the creation of individualized product forms for each patient. This work presents dropwise additive manufacturing of pharmaceutical products (DAMPP), an automated, controlled mini-manufacturing method that deposits active pharmaceutical ingredients (APIs) directly onto edible substrates using drop-on-demand (DoD) inkjet printing technology. The use of DoD technology allows for precise control over the material properties, drug solid state form, drop size, and drop dynamics and can be beneficial in the creation of high-potency drug forms, combination drugs with multiple APIs or individualized medicine products tailored to a specific patient. In this work, DAMPP was used to create dosage forms from solvent-based formulations consisting of API, polymer, and solvent carrier. The forms were then analyzed to determine the reproducibility of creating an on-target dosage form, the morphology of the API of the final form and the dissolution behavior of the drug over time. DAMPP is found to be a viable alternative to traditional mass-manufacturing methods for solvent-based oral dosage forms. © 2013 Wiley Periodicals, Inc. and the American Pharmacists Association.

  11. Preliminary assessment of nine waste-form products/processes for immobilizing transuranic wastes

    Crisler, L.R.

    1980-09-01

    Nine waste-form processes for reduction of the present and projected Transuranic (TRU) waste inventory to an immobilized product have been evaluated. Product formulations, selected properties, preparation methods, technology status, problem areas needing resolution and location of current research development being pursued in the United States are discussed for each process. No definitive utility ranking is attempted due to the early stage of product/process development for TRU waste containing products and the uncertainties in the state of current knowledge of TRU waste feed compositional and quantitative makeup. Of the nine waste form products/processes included in this discussion, bitumen and cements (encapsulation agents) demonstrate the degree of flexibility necessary to immobilize the wide composition range present in the TRU waste inventory. A demonstrated process called Slagging Pyrolysis Incineration converts a varied compositional feed (municipal wastes) to a ''basalt'' like product. This process/product appears to have potential for TRU waste immobilization. The remaining waste forms (borosilicate glass, high-silica glass, glass ceramics, ''SYNROC B'' and cermets) have potential for immobilizing a smaller fraction of the TRU waste inventory than the above discussed waste forms

  12. Computational metabolic engineering strategies for growth-coupled biofuel production by Synechocystis

    Kiyan Shabestary

    2016-12-01

    Full Text Available Chemical and fuel production by photosynthetic cyanobacteria is a promising technology but to date has not reached competitive rates and titers. Genome-scale metabolic modeling can reveal limitations in cyanobacteria metabolism and guide genetic engineering strategies to increase chemical production. Here, we used constraint-based modeling and optimization algorithms on a genome-scale model of Synechocystis PCC6803 to find ways to improve productivity of fermentative, fatty-acid, and terpene-derived fuels. OptGene and MOMA were used to find heuristics for knockout strategies that could increase biofuel productivity. OptKnock was used to find a set of knockouts that led to coupling between biofuel and growth. Our results show that high productivity of fermentation or reversed beta-oxidation derived alcohols such as 1-butanol requires elimination of NADH sinks, while terpenes and fatty-acid based fuels require creating imbalances in intracellular ATP and NADPH production and consumption. The FBA-predicted productivities of these fuels are at least 10-fold higher than those reported so far in the literature. We also discuss the physiological and practical feasibility of implementing these knockouts. This work gives insight into how cyanobacteria could be engineered to reach competitive biofuel productivities. Keywords: Cyanobacteria, Modeling, Flux balance analysis, Biofuel, MOMA, OptFlux, OptKnock

  13. Effect of some metabolic inhibitors on citric acid production Aspergillus niger

    Agrawal, P.K.; Bhatt, C.S.; Viswanathan, L.

    1983-09-01

    Stationary cultures of Aspergillus niger grown on a synthetic medium have been used to study the effect of some metabolic inhibitors on citric acid production. Addition of 0.05 to 1 mM sodium malonate or 0.01 to 0.1 mM potassium ferricyanide, iodoacetate, sodium azide, soldium arsenate or sodium fluoride stimulated citric acid production (3.6 to 45%), but not total titratable acids. Addition of higher concentrations (0.2 to 10 mM) of later inhibitors caused a marked inhibition of fungal growth and citric acid production. The implications of these preliminary findings are discussed. (Refs. 25).

  14. Metabolism of pharmaceutical and personal care products by carrot cell cultures.

    Wu, Xiaoqin; Fu, Qiuguo; Gan, Jay

    2016-04-01

    With the increasing use of treated wastewater and biosolids in agriculture, residues of pharmaceutical and personal care products (PPCPs) in these reused resources may contaminate food produce via plant uptake, constituting a route for human exposure. Although various PPCPs have been reported to be taken up by plants in laboratories or under field conditions, at present little information is available on their metabolism in plants. In this study, we applied carrot cell cultures to investigate the plant metabolism of PPCPs. Five phase I metabolites of carbamazepine were identified and the potential metabolism pathways of carbamazepine were proposed. We also used the carrot cell cultures as a rapid screening tool to initially assess the metabolism potentials of 18 PPCPs. Eleven PPCPs, including acetaminophen, caffeine, meprobamate, primidone, atenolol, trimethoprim, DEET, carbamazepine, dilantin, diazepam, and triclocarban, were found to be recalcitrant to metabolism. The other 7 PPCPs, including triclosan, naproxen, diclofenac, ibuprofen, gemfibrozil, sulfamethoxazole, and atorvastatin, displayed rapid metabolism, with 0.4-47.3% remaining in the culture at the end of the experiment. Further investigation using glycosidase hydrolysis showed that 1.3-20.6% of initially spiked naproxen, diclofenac, ibuprofen, and gemfibrozil were transformed into glycoside conjugates. Results from this study showed that plant cell cultures may be a useful tool for initially exploring the potential metabolites of PPCPs in plants as well as for rapidly screening the metabolism potentials of a variety of PPCPs or other emerging contaminants, and therefore may be used for prioritizing compounds for further comprehensive evaluations. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. UV light selectively coinduces supply pathways from primary metabolism and flavonoid secondary product formation in parsley

    Logemann, Elke; Tavernaro, Annette; Schulz, Wolfgang; Somssich, Imre E.; Hahlbrock, Klaus

    2000-01-01

    The UV light-induced synthesis of UV-protective flavonoids diverts substantial amounts of substrates from primary metabolism into secondary product formation and thus causes major perturbations of the cellular homeostasis. Results from this study show that the mRNAs encoding representative enzymes from various supply pathways are coinduced in UV-irradiated parsley cells (Petroselinum crispum) with two mRNAs of flavonoid glycoside biosynthesis, encoding phenylalanine ammonia-lyase and chalcone synthase. Strong induction was observed for mRNAs encoding glucose 6-phosphate dehydrogenase (carbohydrate metabolism, providing substrates for the shikimate pathway), 3-deoxyarabinoheptulosonate 7-phosphate synthase (shikimate pathway, yielding phenylalanine), and acyl-CoA oxidase (fatty acid degradation, yielding acetyl-CoA), and moderate induction for an mRNA encoding S-adenosyl-homocysteine hydrolase (activated methyl cycle, yielding S-adenosyl-methionine for B-ring methylation). Ten arbitrarily selected mRNAs representing various unrelated metabolic activities remained unaffected. Comparative analysis of acyl-CoA oxidase and chalcone synthase with respect to mRNA expression modes and gene promoter structure and function revealed close similarities. These results indicate a fine-tuned regulatory network integrating those functionally related pathways of primary and secondary metabolism that are specifically required for protective adaptation to UV irradiation. Although the response of parsley cells to UV light is considerably broader than previously assumed, it contrasts greatly with the extensive metabolic reprogramming observed previously in elicitor-treated or fungus-infected cells. PMID:10677554

  16. Retinoid production using metabolically engineered Escherichia coli with a two-phase culture system.

    Jang, Hui-Jeong; Yoon, Sang-Hwal; Ryu, Hee-Kyung; Kim, Jung-Hun; Wang, Chong-Long; Kim, Jae-Yean; Oh, Deok-Kun; Kim, Seon-Won

    2011-07-29

    Retinoids are lipophilic isoprenoids composed of a cyclic group and a linear chain with a hydrophilic end group. These compounds include retinol, retinal, retinoic acid, retinyl esters, and various derivatives of these structures. Retinoids are used as cosmetic agents and effective pharmaceuticals for skin diseases. Retinal, an immediate precursor of retinoids, is derived by β-carotene 15,15'-mono(di)oxygenase (BCM(D)O) from β-carotene, which is synthesized from the isoprenoid building blocks isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Retinoids are chemically unstable and biologically degraded via retinoic acid. Although extensive studies have been performed on the microbial production of carotenoids, retinoid production using microbial metabolic engineering has not been reported. Here, we report retinoid production using engineered Escherichia coli that express exogenous BCM(D)O and the mevalonate (MVA) pathway for the building blocks synthesis in combination with a two-phase culture system using a dodecane overlay. Among the BCM(D)O tested in E. coli, the synthetic retinoid synthesis protein (SR), based on bacteriorhodopsin-related protein-like homolog (Blh) of the uncultured marine bacteria 66A03, showed the highest β-carotene cleavage activity with no residual intracellular β-carotene. By introducing the exogenous MVA pathway, 8.7 mg/L of retinal was produced, which is 4-fold higher production than that of augmenting the MEP pathway (dxs overexpression). There was a large gap between retinal production and β-carotene consumption using the exogenous MVA pathway; therefore, the retinal derivatives were analyzed. The derivatives, except for retinoic acid, that formed were identified, and the levels of retinal, retinol, and retinyl acetate were measured. Amounts as high as 95 mg/L retinoids were obtained from engineered E. coli DH5α harboring the synthetic SR gene and the exogenous MVA pathway in addition to dxs overexpression, which

  17. Using event related potentials to identify a user's behavioural intention aroused by product form design.

    Ding, Yi; Guo, Fu; Zhang, Xuefeng; Qu, Qingxing; Liu, Weilin

    2016-07-01

    The capacity of product form to arouse user's behavioural intention plays a decisive role in further user experience, even in purchase decision, while traditional methods rarely give a fully understanding of user experience evoked by product form, especially the feeling of anticipated use of product. Behavioural intention aroused by product form designs has not yet been investigated electrophysiologically. Hence event related potentials (ERPs) were applied to explore the process of behavioural intention when users browsed different smart phone form designs with brand and price not taken into account for mainly studying the brain activity evoked by variety of product forms. Smart phone pictures with different anticipated user experience were displayed with equiprobability randomly. Participants were asked to click the left mouse button when certain picture gave them a feeling of behavioural intention to interact with. The brain signal of each participant was recorded by Curry 7.0. The results show that pictures with an ability to arouse participants' behavioural intention for further experience can evoke enhanced N300 and LPPs (late positive potentials) in central-parietal, parietal and occipital regions. The scalp topography shows that central-parietal, parietal and occipital regions are more activated. The results indicate that the discrepancy of ERPs can reflect the neural activities of behavioural intention formed or not. Moreover, amplitude of ERPs occurred in corresponding brain areas can be used to measure user experience. The exploring of neural correlated with behavioural intention provide an accurate measurement method of user's perception and help marketers to know which product can arouse users' behavioural intention, maybe taken as an evaluating indicator of product design. Copyright © 2016 Elsevier Ltd and The Ergonomics Society. All rights reserved.

  18. Hydrodynamics-based functional forms of activity metabolism: a case for the power-law polynomial function in animal swimming energetics.

    Papadopoulos, Anthony

    2009-01-01

    The first-degree power-law polynomial function is frequently used to describe activity metabolism for steady swimming animals. This function has been used in hydrodynamics-based metabolic studies to evaluate important parameters of energetic costs, such as the standard metabolic rate and the drag power indices. In theory, however, the power-law polynomial function of any degree greater than one can be used to describe activity metabolism for steady swimming animals. In fact, activity metabolism has been described by the conventional exponential function and the cubic polynomial function, although only the power-law polynomial function models drag power since it conforms to hydrodynamic laws. Consequently, the first-degree power-law polynomial function yields incorrect parameter values of energetic costs if activity metabolism is governed by the power-law polynomial function of any degree greater than one. This issue is important in bioenergetics because correct comparisons of energetic costs among different steady swimming animals cannot be made unless the degree of the power-law polynomial function derives from activity metabolism. In other words, a hydrodynamics-based functional form of activity metabolism is a power-law polynomial function of any degree greater than or equal to one. Therefore, the degree of the power-law polynomial function should be treated as a parameter, not as a constant. This new treatment not only conforms to hydrodynamic laws, but also ensures correct comparisons of energetic costs among different steady swimming animals. Furthermore, the exponential power-law function, which is a new hydrodynamics-based functional form of activity metabolism, is a special case of the power-law polynomial function. Hence, the link between the hydrodynamics of steady swimming and the exponential-based metabolic model is defined.

  19. Hydrodynamics-based functional forms of activity metabolism: a case for the power-law polynomial function in animal swimming energetics.

    Anthony Papadopoulos

    Full Text Available The first-degree power-law polynomial function is frequently used to describe activity metabolism for steady swimming animals. This function has been used in hydrodynamics-based metabolic studies to evaluate important parameters of energetic costs, such as the standard metabolic rate and the drag power indices. In theory, however, the power-law polynomial function of any degree greater than one can be used to describe activity metabolism for steady swimming animals. In fact, activity metabolism has been described by the conventional exponential function and the cubic polynomial function, although only the power-law polynomial function models drag power since it conforms to hydrodynamic laws. Consequently, the first-degree power-law polynomial function yields incorrect parameter values of energetic costs if activity metabolism is governed by the power-law polynomial function of any degree greater than one. This issue is important in bioenergetics because correct comparisons of energetic costs among different steady swimming animals cannot be made unless the degree of the power-law polynomial function derives from activity metabolism. In other words, a hydrodynamics-based functional form of activity metabolism is a power-law polynomial function of any degree greater than or equal to one. Therefore, the degree of the power-law polynomial function should be treated as a parameter, not as a constant. This new treatment not only conforms to hydrodynamic laws, but also ensures correct comparisons of energetic costs among different steady swimming animals. Furthermore, the exponential power-law function, which is a new hydrodynamics-based functional form of activity metabolism, is a special case of the power-law polynomial function. Hence, the link between the hydrodynamics of steady swimming and the exponential-based metabolic model is defined.

  20. Metabolic and Kinetic analyses of influenza production in perfusion HEK293 cell culture

    Lohr Verena

    2011-09-01

    Full Text Available Abstract Background Cell culture-based production of influenza vaccine remains an attractive alternative to egg-based production. Short response time and high production yields are the key success factors for the broader adoption of cell culture technology for industrial manufacturing of pandemic and seasonal influenza vaccines. Recently, HEK293SF cells have been successfully used to produce influenza viruses, achieving hemagglutinin (HA and infectious viral particle (IVP titers in the highest ranges reported to date. In the same study, it was suggested that beyond 4 × 106 cells/mL, viral production was limited by a lack of nutrients or an accumulation of toxic products. Results To further improve viral titers at high cell densities, perfusion culture mode was evaluated. Productivities of both perfusion and batch culture modes were compared at an infection cell density of 6 × 106 cells/mL. The metabolism, including glycolysis, glutaminolysis and amino acids utilization as well as physiological indicators such as viability and apoptosis were extensively documented for the two modes of culture before and after viral infection to identify potential metabolic limitations. A 3 L bioreactor with a perfusion rate of 0.5 vol/day allowed us to reach maximal titers of 3.3 × 1011 IVP/mL and 4.0 logHA units/mL, corresponding to a total production of 1.0 × 1015 IVP and 7.8 logHA units after 3 days post-infection. Overall, perfusion mode titers were higher by almost one order of magnitude over the batch culture mode of production. This improvement was associated with an activation of the cell metabolism as seen by a 1.5-fold and 4-fold higher consumption rates of glucose and glutamine respectively. A shift in the viral production kinetics was also observed leading to an accumulation of more viable cells with a higher specific production and causing an increase in the total volumetric production of infectious influenza particles. Conclusions These results

  1. Differential proteomic analysis reveals novel links between primary metabolism and antibiotic production in Amycolatopsis balhimycina

    Gallo, G.; Renzone, G.; Alduina, R.

    2010-01-01

    A differential proteomic analysis, based on 2-DE and MS procedures, was performed on Amycolatopsis balhimycina DSM5908, the actinomycete producing the vancomycin-like antibiotic balhimycin. A comparison of proteomic profiles before and during balhimycin production characterized differentially...... available over the World Wide Web as interactive web pages (http://www.unipa.it/ampuglia/Abal-proteome-maps). Functional clustering analysis revealed that differentially expressed proteins belong to functional groups involved in central carbon metabolism, amino acid metabolism and protein biosynthesis...... intermediates, were upregulated during antibiotic production. qRT-PCR analysis revealed that 8 out of 14 upregulated genes showed a positive correlation between changes at translational and transcriptional expression level. Furthermore, proteomic analysis of two nonproducing mutants, restricted to a sub...

  2. Production of the sesquiterpenoid (+)-nootkatone by metabolic engineering of Pichia pastoris.

    Wriessnegger, Tamara; Augustin, Peter; Engleder, Matthias; Leitner, Erich; Müller, Monika; Kaluzna, Iwona; Schürmann, Martin; Mink, Daniel; Zellnig, Günther; Schwab, Helmut; Pichler, Harald

    2014-07-01

    The sesquiterpenoid (+)-nootkatone is a highly demanded and highly valued aroma compound naturally found in grapefruit, pummelo or Nootka cypress tree. Extraction of (+)-nootkatone from plant material or its production by chemical synthesis suffers from low yields and the use of environmentally harmful methods, respectively. Lately, major attention has been paid to biotechnological approaches, using cell extracts or whole-cell systems for the production of (+)-nootkatone. In our study, the yeast Pichia pastoris initially was applied as whole-cell biocatalyst for the production of (+)-nootkatone from (+)-valencene, the abundant aroma compound of oranges. Therefore, we generated a strain co-expressing the premnaspirodiene oxygenase of Hyoscyamus muticus (HPO) and the Arabidopsis thaliana cytochrome P450 reductase (CPR) that hydroxylated extracellularly added (+)-valencene. Intracellular production of (+)-valencene by co-expression of valencene synthase from Callitropsis nootkatensis resolved the phase-transfer issues of (+)-valencene. Bi-phasic cultivations of P. pastoris resulted in the production of trans-nootkatol, which was oxidized to (+)-nootkatone by an intrinsic P. pastoris activity. Additional overexpression of a P. pastoris alcohol dehydrogenase and truncated hydroxy-methylglutaryl-CoA reductase (tHmg1p) significantly enhanced the (+)-nootkatone yield to 208mg L(-1) cell culture in bioreactor cultivations. Thus, metabolically engineered yeast P. pastoris represents a valuable, whole-cell system for high-level production of (+)-nootkatone from simple carbon sources. Copyright © 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  3. Biofuel production in Escherichia coli. The role of metabolic engineering and synthetic biology

    Clomburg, James M. [Rice Univ., Houston, TX (United States). Dept. of Chemical and Biomolecular Engineering; Gonzalez, Ramon [Rice Univ., Houston, TX (United States). Dept. of Chemical and Biomolecular Engineering; Rice Univ., Houston, TX (United States). Dept. of Bioengineering

    2010-03-15

    The microbial production of biofuels is a promising avenue for the development of viable processes for the generation of fuels from sustainable resources. In order to become cost and energy effective, these processes must utilize organisms that can be optimized to efficiently produce candidate fuels from a variety of feedstocks. Escherichia coli has become a promising host organism for the microbial production of biofuels in part due to the ease at which this organism can be manipulated. Advancements in metabolic engineering and synthetic biology have led to the ability to efficiently engineer E. coli as a biocatalyst for the production of a wide variety of potential biofuels from several biomass constituents. This review focuses on recent efforts devoted to engineering E. coli for the production of biofuels, with emphasis on the key aspects of both the utilization of a variety of substrates as well as the synthesis of several promising biofuels. Strategies for the efficient utilization of carbohydrates, carbohydrate mixtures, and noncarbohydrate carbon sources will be discussed along with engineering efforts for the exploitation of both fermentative and nonfermentative pathways for the production of candidate biofuels such as alcohols and higher carbon biofuels derived from fatty acid and isoprenoid pathways. Continued advancements in metabolic engineering and synthetic biology will help improve not only the titers, yields, and productivities of biofuels discussed herein, but also increase the potential range of compounds that can be produced. (orig.)

  4. The Prospect of Using Complete Feed in Goat Production: A Review on its Utility and Physical Form and Animal Responses

    Simon P Ginting

    2009-06-01

    Full Text Available Complete feed is a strategic feeding system that has been widely adopted by the dairy cattle industry, but it has been rarely practised in goat enterprises. The prospect of using complete feed for goat production could be considered from two aspects, namely 1 its relevancy to the goat metabolic requirement, and 2 its potential as an effective means for maximal utilization of crop residues and agro-industrial byproducts as alternative feeds. Metabolically, the higher energy requirement and the lower gut capacity of goats due to its relatively smaller body size make this animal more adapted to feedstuffs with denser nutrient contents as typified by the complete feed. As complete feed is characteristically dry, it has potential to cause hypovolemia and induce later a lower feed consumption when fed to goats. But, this event occurs only during the initial meal and the condition returns to the normal state thereafter. Total saliva secretion tend to decrease by consuming dry feed which can cause parakeratosis, laminitis or acidosis. However, these metabolic disorder could be prevented by formulating complete feed with optimal roughage/concentrate ratio. Review from literatures showed that, when used in complete feed, the inclusion rate of several low palatability crop residues or agro-industrial ranged from 15 to 60%. The roughage/concentrate ratio was in the range of 0.25 to 3.0. Some physical characteristics are important for effective complete feed such as the particle size of roughage, the content of physically effective fibre and the form of the complete feed. Complete feed processed into pellet generally resulted in better performances. The ME and CP content of complete feed used ranged from 1800 to 2800 kcal/kg DM and from 15 to 20%, respectively. The rate of feed intake by goats receiving complete feed ranged from 2.0 to 4.9% BW, the ADG ranged from 40 to 145 g, FCR ranged from 5.2 to 13.0 and DM digestibility ranged from 62 to 81%. These

  5. Ruminant Nutrition Symposium: ruminant production and metabolic responses to heat stress.

    Baumgard, L H; Rhoads, R P

    2012-06-01

    Heat stress compromises efficient animal production by marginalizing nutrition, management, and genetic selection efforts to maximize performance endpoints. Modifying farm infrastructure has yielded modest success in mitigating heat stress-related losses, yet poor production during the summer remains arguably the costliest issue facing livestock producers. Reduced output (e.g., milk yield and muscle growth) during heat stress was traditionally thought to result from decreased nutrient intake (i.e., a classic biological response shared by all animals during environmental-induced hyperthermia). Our recent observations have begun to challenge this belief and indicate heat-stressed animals employ novel homeorhetic strategies to direct metabolic and fuel selection priorities independently of nutrient intake or energy balance. Alterations in systemic physiology support a shift in carbohydrate metabolism, evident by increased basal and stimulated circulating insulin concentrations. Perhaps most intriguing given the energetic shortfall of the heat-stressed animal is the apparent lack of basal adipose tissue mobilization coupled with a reduced responsiveness to lipolytic stimuli. Thus, the heat stress response markedly alters postabsorptive carbohydrate, lipid, and protein metabolism independently of reduced feed intake through coordinated changes in fuel supply and utilization by multiple tissues. Interestingly, the systemic, cellular, and molecular changes appear conserved amongst different species and physiological states. Ultimately, these changes result in the reprioritization of fuel selection during heat stress, which appears to be primarily responsible for reduced ruminant animal productivity during the warm summer months.

  6. Disruption of quercetin metabolism by fungicide affects energy production in honey bees (Apis mellifera).

    Mao, Wenfu; Schuler, Mary A; Berenbaum, May R

    2017-03-07

    Cytochrome P450 monooxygenases (P450) in the honey bee, Apis mellifera , detoxify phytochemicals in honey and pollen. The flavonol quercetin is found ubiquitously and abundantly in pollen and frequently at lower concentrations in honey. Worker jelly consumed during the first 3 d of larval development typically contains flavonols at very low levels, however. RNA-Seq analysis of gene expression in neonates reared for three days on diets with and without quercetin revealed that, in addition to up-regulating multiple detoxifying P450 genes, quercetin is a negative transcriptional regulator of mitochondrion-related nuclear genes and genes encoding subunits of complexes I, III, IV, and V in the oxidative phosphorylation pathway. Thus, a consequence of inefficient metabolism of this phytochemical may be compromised energy production. Several P450s metabolize quercetin in adult workers. Docking in silico of 121 pesticide contaminants of American hives into the active pocket of CYP9Q1, a broadly substrate-specific P450 with high quercetin-metabolizing activity, identified six triazole fungicides, all fungal P450 inhibitors, that dock in the catalytic site. In adults fed combinations of quercetin and the triazole myclobutanil, the expression of five of six mitochondrion-related nuclear genes was down-regulated. Midgut metabolism assays verified that adult bees consuming quercetin with myclobutanil metabolized less quercetin and produced less thoracic ATP, the energy source for flight muscles. Although fungicides lack acute toxicity, they may influence bee health by interfering with quercetin detoxification, thereby compromising mitochondrial regeneration and ATP production. Thus, agricultural use of triazole fungicides may put bees at risk of being unable to extract sufficient energy from their natural food.

  7. Solar photocatalytic oxidation of recalcitrant natural metabolic by-products of amoxicillin biodegradation.

    Pereira, João H O S; Reis, Ana C; Homem, Vera; Silva, José A; Alves, Arminda; Borges, Maria T; Boaventura, Rui A R; Vilar, Vítor J P; Nunes, Olga C

    2014-11-15

    The contamination of the aquatic environment by non-metabolized and metabolized antibiotic residues has brought the necessity of alternative treatment steps to current water decontamination technologies. This work assessed the feasibility of using a multistage treatment system for amoxicillin (AMX) spiked solutions combining: i) a biological treatment process using an enriched culture to metabolize AMX, with ii) a solar photocatalytic system to achieve the removal of the metabolized transformation products (TPs) identified via LC-MS, recalcitrant to further biological degradation. Firstly, a mixed culture (MC) was obtained through the enrichment of an activated sludge sample collected in an urban wastewater treatment plant (WWTP). Secondly, different aqueous matrices spiked with AMX were treated with the MC and the metabolic transformation products were identified. Thirdly, the efficiency of two solar assisted photocatalytic processes (TiO2/UV or Fe(3+)/Oxalate/H2O2/UV-Vis) was assessed in the degradation of the obtained TPs using a lab-scale prototype photoreactor equipped with a compound parabolic collector (CPC). Highest AMX specific biodegradation rates were obtained in buffer and urban wastewater (WW) media (0.10 ± 0.01 and 0.13 ± 0.07 g(AMX) g(biomass)(-1) h(-1), respectively). The resulting TPs, which no longer presented antibacterial activity, were identified as amoxicilloic acid (m/z = 384). The performance of the Fe(3+)/Oxalate/H2O2/UV-Vis system in the removal of the TPs from WW medium was superior to the TiO2/UV process (TPs no longer detected after 40 min (QUV = 2.6 kJ L(-1)), against incomplete TPs removal after 240 min (QUV = 14.9 kJ L(-1)), respectively). Copyright © 2014 Elsevier Ltd. All rights reserved.

  8. Deletion of Type I glutamine synthetase deregulates nitrogen metabolism and increases ethanol production in Clostridium thermocellum

    Rydzak, Thomas [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Garcia, David [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Stevenson, David M. [Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology; Sladek, Margaret [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Klingeman, Dawn M. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Holwerda, Evert K. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division; Dartmouth College, Hanover, NH (United States). Thayer School of Engineering; Amador-Noguez, Daniel [Univ. of Wisconsin, Madison, WI (United States). Dept. of Bacteriology; Brown, Steven D. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center; Guss, Adam M. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Biosciences Division, BioEnergy Science Center

    2017-05-01

    Clostridium thermocellum rapidly deconstructs cellulose and ferments resulting hydrolysis products into ethanol and other products, and is thus a promising platform organism for the development of cellulosic biofuel production via consolidated bioprocessing. And while recent metabolic engineering strategies have targeted eliminating canonical fermentation products (acetate, lactate, formate, and H2), C. thermocellum also secretes amino acids, which has limited ethanol yields in engineered strains to approximately 70% of the theoretical maximum. To decrease amino acid secretion, we attempted to reduce ammonium assimilation by deleting the Type I glutamine synthetase (glnA) in C. thermocellum. Deletion of glnA reduced levels of secreted valine and total amino acids by 53% and 44% respectively, and increased ethanol yields by 53%. RNA-seq analysis revealed that genes encoding the RNF-complex were more highly expressed in ΔglnA and may have a role in improving NADH-availability for ethanol production. While a significant up-regulation of genes involved in nitrogen assimilation and urea uptake suggested that deletion of glnA induces a nitrogen starvation response, metabolomic analysis showed an increase in intracellular glutamine and α-ketoglutarate levels indicative of nitrogen-rich conditions. Here, we propose that deletion of glnA causes deregulation of nitrogen metabolism, leading to overexpression of nitrogen metabolism genes and, in turn, elevated glutamine/α-ketoglutarate levels. Here we demonstrate that perturbation of nitrogen assimilation is a promising strategy to redirect flux from the production of nitrogenous compounds toward biofuels in C. thermocellum.

  9. Systems-wide metabolic pathway engineering in Corynebacterium glutamicum for bio-based production of diaminopentane.

    Kind, Stefanie; Jeong, Weol Kyu; Schröder, Hartwig; Wittmann, Christoph

    2010-07-01

    In the present work the Gram-positive bacterium Corynebacterium glutamicum was engineered into an efficient, tailor-made production strain for diaminopentane (cadaverine), a highly attractive building block for bio-based polyamides. The engineering comprised expression of lysine decarboxylase (ldcC) from Escherichia coli, catalyzing the conversion of lysine into diaminopentane, and systems-wide metabolic engineering of central supporting pathways. Substantially re-designing the metabolism yielded superior strains with desirable properties such as (i) the release from unwanted feedback regulation at the level of aspartokinase and pyruvate carboxylase by introducing the point mutations lysC311 and pycA458, (ii) an optimized supply of the key precursor oxaloacetate by amplifying the anaplerotic enzyme, pyruvate carboxylase, and deleting phosphoenolpyruvate carboxykinase which otherwise removes oxaloacetate, (iii) enhanced biosynthetic flux via combined amplification of aspartokinase, dihydrodipicolinate reductase, diaminopimelate dehydrogenase and diaminopimelate decarboxylase, and (iv) attenuated flux into the threonine pathway competing with production by the leaky mutation hom59 in the homoserine dehydrogenase gene. Lysine decarboxylase proved to be a bottleneck for efficient production, since its in vitro activity and in vivo flux were closely correlated. To achieve an optimal strain having only stable genomic modifications, the combination of the strong constitutive C. glutamicum tuf promoter and optimized codon usage allowed efficient genome-based ldcC expression and resulted in a high diaminopentane yield of 200 mmol mol(-1). By supplementing the medium with 1 mgL(-1) pyridoxal, the cofactor of lysine decarboxylase, the yield was increased to 300 mmol mol(-1). In the production strain obtained, lysine secretion was almost completely abolished. Metabolic analysis, however, revealed substantial formation of an as yet unknown by-product. It was identified as an

  10. Deletion of Type I glutamine synthetase deregulates nitrogen metabolism and increases ethanol production in Clostridium thermocellum.

    Rydzak, Thomas; Garcia, David; Stevenson, David M; Sladek, Margaret; Klingeman, Dawn M; Holwerda, Evert K; Amador-Noguez, Daniel; Brown, Steven D; Guss, Adam M

    2017-05-01

    Clostridium thermocellum rapidly deconstructs cellulose and ferments resulting hydrolysis products into ethanol and other products, and is thus a promising platform organism for the development of cellulosic biofuel production via consolidated bioprocessing. While recent metabolic engineering strategies have targeted eliminating canonical fermentation products (acetate, lactate, formate, and H 2 ), C. thermocellum also secretes amino acids, which has limited ethanol yields in engineered strains to approximately 70% of the theoretical maximum. To investigate approaches to decrease amino acid secretion, we attempted to reduce ammonium assimilation by deleting the Type I glutamine synthetase (glnA) in an essentially wild type strain of C. thermocellum. Deletion of glnA reduced levels of secreted valine and total amino acids by 53% and 44% respectively, and increased ethanol yields by 53%. RNA-seq analysis revealed that genes encoding the RNF-complex were more highly expressed in ΔglnA and may have a role in improving NADH-availability for ethanol production. While a significant up-regulation of genes involved in nitrogen assimilation and urea uptake suggested that deletion of glnA induces a nitrogen starvation response, metabolomic analysis showed an increase in intracellular glutamine levels indicative of nitrogen-rich conditions. We propose that deletion of glnA causes deregulation of nitrogen metabolism, leading to overexpression of nitrogen metabolism genes and, in turn, elevated glutamine levels. Here we demonstrate that perturbation of nitrogen assimilation is a promising strategy to redirect flux from the production of nitrogenous compounds toward biofuels in C. thermocellum. Copyright © 2017. Published by Elsevier Inc.

  11. The RNA chaperone Hfq impacts growth, metabolism and production of virulence factors in Yersinia enterocolitica.

    Tamara Kakoschke

    Full Text Available To adapt to changes in environmental conditions, bacteria regulate their gene expression at the transcriptional but also at the post-transcriptional level, e.g. by small RNAs (sRNAs which modulate mRNA stability and translation. The conserved RNA chaperone Hfq mediates the interaction of many sRNAs with their target mRNAs, thereby playing a global role in fine-tuning protein production. In this study, we investigated the significance of Hfq for the enteropathogen Yersina enterocolitica serotype O:8. Hfq facilitated optimal growth in complex and minimal media. Our comparative protein analysis of parental and hfq-negative strains suggested that Hfq promotes lipid metabolism and transport, cell redox homeostasis, mRNA translation and ATP synthesis, and negatively affects carbon and nitrogen metabolism, transport of siderophore and peptides and tRNA synthesis. Accordingly, biochemical tests indicated that Hfq represses ornithine decarboxylase activity, indole production and utilization of glucose, mannitol, inositol and 1,2-propanediol. Moreover, Hfq repressed production of the siderophore yersiniabactin and its outer membrane receptor FyuA. In contrast, hfq mutants exhibited reduced urease production. Finally, strains lacking hfq were more susceptible to acidic pH and oxidative stress. Unlike previous reports in other Gram-negative bacteria, Hfq was dispensable for type III secretion encoded by the virulence plasmid. Using a chromosomally encoded FLAG-tagged Hfq, we observed increased production of Hfq-FLAG in late exponential and stationary phases. Overall, Hfq has a profound effect on metabolism, resistance to stress and modulates the production of two virulence factors in Y. enterocolitica, namely urease and yersiniabactin.

  12. Clean forming of stainless steel and titanium products by lubricious oxides

    Heikkilä, Irma; Wadman, Boel; Thoors, Håkan

    2012-01-01

    to industrial forming processes. Preliminary evaluations show a beneficial influence of two oxides types, on stainless steel and on titanium. More work is needed to test the lubricating effect in other forming operations and to analyse the sustainability aspects for products manufactured with this alternative......Big social benefits can be attained through increased use of stainless steel or titanium in new sheet metal applications. Unfortunately, forming of these materials is often a challenging and costly operation, that can lead to environmental and health problems when solving the technical limitations...

  13. Waste acceptance product specifications for vitrified high-level waste forms

    Applewhite-Ramsey, A.; Sproull, J.F.

    1993-01-01

    The Nuclear Waste Policy Act of 1982 mandated that all high-level waste (HLW) be sent to a federal geologic repository for permanent disposal. DOE published the Environmental Assessment in 1982 which identified borosilicate glass as the chosen HLW form. 1 In 1985 the Department of Energy instituted a Waste Acceptance Process to assure that DWPF glass waste forms would be acceptable to such a repository. This assurance was important since production of waste forms will precede repository construction and licensing. As part of this Waste Acceptance Process, the DOE Office of Civilian Radioactive Waste Management (RW) formed the Waste Acceptance Committee (WAC). The WAC included representatives from the candidate repository sites, the waste producing sites and DOE. The WAC was responsible for developing the Waste Acceptance Preliminary Specifications (WAPS) which defined the requirements the waste forms must meet to be compatible with the candidate repository geologies

  14. Metabolic engineering of Saccharomyces cerevisiae for production of germacrene A, a precursor of beta-elemene

    Hu, Yating; Zhou, Yongjin J.; Bao, Jichen

    2017-01-01

    inefficient and suffers from limited natural resources. Here, we engineered a yeast cell factory for the sustainable production of germacrene A, which can be transformed to beta-elemene by a one-step chemical reaction in vitro. Two heterologous germacrene A synthases (GASs) converting farnesyl pyrophosphate...... (FPP) to germacrene A were evaluated in yeast for their ability to produce germacrene A. Thereafter, several metabolic engineering strategies were used to improve the production level. Overexpression of truncated 3-hydroxyl-3-methylglutaryl-CoA reductase and fusion of FPP synthase with GAS, led...

  15. The relationship of metabolic burden to productivity levels in CHO cell lines.

    Zou, Wu; Edros, Raihana; Al-Rubeai, Mohamed

    2018-03-01

    The growing demand for recombinant therapeutics has driven biotechnologists to develop new production strategies. One such strategy for increasing the expression of heterologous proteins has focused on enhancing cell-specific productivity through environmental perturbations. In this work, the effects of hypothermia, hyperosmolarity, high shear stress, and sodium butyrate treatment on growth and productivity were studied using three (low, medium, and high producing) CHO cell lines that differed in their specific productivities of monoclonal antibody. In all three cell lines, the inhibitory effect of these parameters on proliferation was demonstrated. Additionally, compared to the control, specific productivity was enhanced under all conditions and exhibited a consistent cell line specific pattern, with maximum increases (50-290%) in the low producer, and minimum increases (7-20%) in the high producer. Thus, the high-producing cell line was less responsive to environmental perturbations than the low-producing cell line. We hypothesize that this difference is most likely due to the bottleneck associated with a higher metabolic burden caused by higher antibody expression. Increased recombinant mRNA levels and pyruvate carboxylase activities due to low temperature and hyperosmotic stress were found to be positively associated with the metabolic burden. © 2017 International Union of Biochemistry and Molecular Biology, Inc.

  16. X-ray diffraction study of slags forming during corrosion resistant steel production

    Slavov, V.I.; Zadorozhnaya, V.N.; Shurygina, A.V.

    1990-01-01

    Using X-ray diffraction analysis slags, forming during corrosion-resistant 12Kh18N10T grade steel production by two flowsheets, are studied. Standard two-slag technology of steel production does not provide efficient disintegration of chromospinelides in slags, gives high steel contamination with respect to nonmetallic impurities, coarse structure and, as a consequence, presence of macrodefects on rolled products surface. One-slag steel melting technology with titanium alloying of the steel at vacuum causes fast removal of chromospinelides at the beginning of reduction period, promotes titanium absorption by the steel, refines nonmetallic inclusions, provides more fine structure and steel plasticity, removes surface defects

  17. Off-Line Testing of Tribo-Systems for Sheet Metal Forming Production

    Bay, Niels; Ceron, Ermanno

    2014-01-01

    Off-line testing of new tribo-systems for sheet metal forming production is an important issue, when new, environmentally benign lubricants are to be introduced. To obtain useful results it is, however, vital to ensure similar conditions as in the production process regarding the main tribo...... leading to very high tool/workpiece interface pressure and temperature in the second re-draw. Under such conditions only the best lubricant systems work satisfactory, and the paper shows how the performance of different tribo-systems in production may be predicted by off-line testing combined...

  18. Elaboration of the technology of forming a conical product of sheet metal

    W. Matysiak

    2010-01-01

    Full Text Available The work presents a general knowledge about spinning draw pieces of sheets, one of multi-operational processes of spinning a sheet metal conical product without machining. The objective of the work was to elaborate both the technology of forming conical products of sheet metal and execution of technological tests as well as to determine the technological parameters for the process of spinning a conical insert. As a result of the investigations, the products with improved mechanical properties, stricter execution tolerance and low roughness have been obtained. The series of 200 prototype conical inserts for the shipbuilding industry have been made.

  19. Effects of dietary bread crust Maillard reaction products on calcium and bone metabolism in rats.

    Roncero-Ramos, Irene; Delgado-Andrade, Cristina; Haro, Ana; Ruiz-Roca, Beatriz; Morales, Francisco J; Navarro, María Pilar

    2013-06-01

    Maillard reaction products (MRP) consumption has been related with the development of bone degenerative disorders, probably linked to changes in calcium metabolism. We aimed to investigate the effects of MRP intake from bread crust on calcium balance and its distribution, and bone metabolism. During 88 days, rats were fed control diet or diets containing bread crust as source of MRP, or its soluble high molecular weight, soluble low molecular weight or insoluble fractions (bread crust, HMW, LMW and insoluble diets, respectively). In the final week, a calcium balance was performed, then animals were sacrified and some organs removed to analyse calcium levels. A second balance was carried out throughout the experimental period to calculate global calcium retention. Biochemical parameters and bone metabolism markers were measured in serum or urine. Global calcium bioavailability was unmodified by consumption of bread crust or its isolate fractions, corroborating the previously described low affinity of MRP to bind calcium. Despite this, a higher calcium concentration was found in femur due to smaller bones having a lower relative density. The isolate consumption of the fractions altered some bone markers, reflecting a situation of increased bone resorption or higher turnover; this did not take place in the animals fed the bread crust diet. Thus, the bread crust intake does not affect negatively calcium bioavailability and bone metabolism.

  20. Metabolic engineering of Cyanobacteria and microalgae for enhanced production of biofuels and high-value products.

    Gomaa, M A; Al-Haj, L; Abed, R M M

    2016-10-01

    A lot of research has been performed on Cyanobacteria and microalgae with the aim to produce numerous biotechnological products. However, native strains have a few shortcomings, like limitations in cultivation, harvesting and product extraction, which prevents reaching optimal production value at lowest costs. Such limitations require the intervention of genetic engineering to produce strains with superior properties. Promising advancements in the cultivation of Cyanobacteria and microalgae have been achieved by improving photosynthetic efficiency through increasing RuBisCO activity and truncation of light-harvesting antennae. Genetic engineering has also contributed to final product extraction by inducing autolysis and product secretory systems, to enable direct product recovery without going through costly extraction steps. In this review, we summarize the different enzymes and pathways that have been targeted thus far for improving cultivation aspects, harvesting and product extraction in Cyanobacteria and microalgae. With synthetic biology advancements, genetically engineered strains can be generated to resolve demanding process issues and achieve economic practicality. This comprehensive overview of gene modifications will be useful to researchers in the field to employ on their strains to increase their yields and improve the economic feasibility of the production process. © 2016 The Society for Applied Microbiology.

  1. 78 FR 21912 - Proposed Information Collection; Comment Request; Processed Products Family of Forms

    2013-04-12

    ... Collection; Comment Request; Processed Products Family of Forms AGENCY: National Oceanic and Atmospheric... NOAA in the economic and social analyses developed when proposing and evaluating fishery management... collection). Affected Public: Business or other for-profit organizations. Estimated Number of Respondents...

  2. Simple product-form bounds for queueing networks with finite clusters

    van Dijk, N.M.; van der Sluis, E.

    2001-01-01

    Queueing networks are studied with finite capacity constraints for clusters of stations. First, by an instructive tandem cluster example it is shown how a product-form modification method for networks with finite stations can be extended to networks with finite clusters. Next, a general result is

  3. Focus on Form, Learner Uptake and Subsequent Lexical Gains in Learners' Oral Production

    Alcon-Soler, Eva

    2009-01-01

    This descriptive study reports findings on the relationship between focus on form, learner uptake and subsequent lexical gains in learners' oral production. The data for the study consisted in 17 45-minute audio-recorded teacher-led conversations, 204 learners' diaries (17 sessions x 12 learners) reporting what they had learned after each…

  4. Biogenic amine content, histamine-forming bacteria, and adulteration of pork in tuna sausage products.

    Kung, Hsien-Feng; Tsai, Yung-Hsiang; Chang, Shih-Chih; Hong, Tang-Yao

    2012-10-01

    Twenty-five tuna sausage products were purchased from retail markets in Taiwan. The rates of occurrence of biogenic amines, histamine-forming bacteria, and adulteration by pork and poultry were determined. The average content of various biogenic amines in all tested samples was less than 2.0 mg/100 g (Makaira nigricans (blue marlin).

  5. On the structure of the space of geometric product-form models

    Bayer, Nimrod; Boucherie, Richardus J.

    2002-01-01

    This article deals with Markovian models defined on a finite-dimensional discrete state space and possess a stationary state distribution of a product-form. We view the space of such models as a mathematical object and explore its structure. We focus on models on an orthant [script Z]+n, which are

  6. Improved Production of a Heterologous Amylase in Saccharomyces cerevisiae by Inverse Metabolic Engineering

    Liu, Zihe; Liu, Lifang; Osterlund, Tobias

    2014-01-01

    this modification alone, the amylase secretion could be improved by 35%. As a complement to the identification of genomic variants, transcriptome analysis was also performed in order to understand on a global level the transcriptional changes associated with the improved amylase production caused by UV mutagenesis.......The increasing demand for industrial enzymes and biopharmaceutical proteins relies on robust production hosts with high protein yield and productivity. Being one of the best-studied model organisms and capable of performing posttranslational modifications, the yeast Saccharomyces cerevisiae...... is widely used as a cell factory for recombinant protein production. However, many recombinant proteins are produced at only 1% (or less) of the theoretical capacity due to the complexity of the secretory pathway, which has not been fully exploited. In this study, we applied the concept of inverse metabolic...

  7. Metabolic engineering of Escherichia coli for limonene and perillyl alcohol production.

    Alonso-Gutierrez, Jorge; Chan, Rossana; Batth, Tanveer S; Adams, Paul D; Keasling, Jay D; Petzold, Christopher J; Lee, Taek Soon

    2013-09-01

    Limonene is a valuable monoterpene used in the production of several commodity chemicals and medicinal compounds. Among them, perillyl alcohol (POH) is a promising anti-cancer agent that can be produced by hydroxylation of limonene. We engineered E. coli with a heterologous mevalonate pathway and limonene synthase for production of limonene followed by coupling with a cytochrome P450, which specifically hydroxylates limonene to produce POH. A strain containing all mevalonate pathway genes in a single plasmid produced limonene at titers over 400mg/L from glucose, substantially higher than has been achieved in the past. Incorporation of a cytochrome P450 to hydroxylate limonene yielded approximately 100mg/L of POH. Further metabolic engineering of the pathway and in situ product recovery using anion exchange resins would make this engineered E. coli a potential production platform for any valuable limonene derivative. © 2013 Elsevier Inc. All rights reserved.

  8. Proteomic analysis of the metabolic adaptation of the biocontrol agent Pseudozyma flocculosa leading to glycolipid production

    Bélanger Richard R

    2010-02-01

    Full Text Available Abstract The yeast-like epiphytic fungus Pseudozyma flocculosa is known to antagonize powdery mildew fungi through proliferation on colonies presumably preceded by the release of an antifungal glycolipid (flocculosin. In culture conditions, P. flocculosa can be induced to produce or not flocculosin through manipulation of the culture medium nutrients. In order to characterize and understand the metabolic changes in P. flocculosa linked to glycolipid production, we conducted a 2-DE proteomic analysis and compared the proteomic profile of P. flocculosa growing under conditions favoring the development of the fungus (control or conducive to flocculosin synthesis (stress. A large number of protein spots (771 were detected in protein extracts of the control treatment compared to only 435 matched protein spots in extracts of the stress cultures, which clearly suggests an important metabolic reorganization in slow-growing cells producing flocculosin. From the latter treatment, we were able to identify 21 protein spots that were either specific to the treatment or up-regulated significantly (2-fold increase. All of them were identified based on similarity between predicted ORF of the newly sequenced genome of P. flocculosa with Ustilago maydis' available annotated sequences. These proteins were associated with the carbon and fatty acid metabolism, and also with the filamentous change of the fungus leading to flocculosin production. This first look into the proteome of P. flocculosa suggests that flocculosin synthesis is elicited in response to specific stress or limiting conditions.

  9. Correlation Between Metabolic Syndrome, Periodontitis and Reactive Oxygen Species Production. A Pilot Study.

    Patini, Romeo; Gallenzi, Patrizia; Spagnuolo, Gianrico; Cordaro, Massimo; Cantiani, Monica; Amalfitano, Adriana; Arcovito, Alessandro; Callà, Cinzia; Mingrone, Gertrude; Nocca, Giuseppina

    2017-01-01

    Metabolic syndrome (MetS) is associated with an increased risk of periodontitis even if the mechanism is unknown. Since both MetS and periodontitis are characterized by an alteration of inflammation status, the aim of this pilot study was to determine if differences in ROS metabolism of phagocytes isolated from (A) patients with MetS, (B) patients with both MetS and mild periodontitis, (C) healthy subjects and (D) normal weight subjects with mild periodontitis, were present. ROS metabolism was studied by a Chemiluminescence (CL) technique: the system was made up of luminol (100 nmol/L) and cells (1 × 10 5 ) in the presence or absence of stimulus constituted by opsonized zymosan (0.5 mg). The final volume (1.0 mL) was obtained using modified KRP buffer. ROS production was measured at 25°C for 2 h, using an LB 953 luminometer (Berthold, EG & G Co, Germany). All the experiments were performed in triplicate. All results are mean ± standard deviation (SD). The group of means was compared by the analysis of variance "(ANOVA)". A value of p < 0.05 was considered significant. Results showed that basal ROS production (both from PMNs and from PBMs) of groups A, B and D was increased with respect to that obtained from group C ( p <0.05). These results are congruent with literature data, although the actual clinical relevance of the phenomenon remains to be evaluated.

  10. Selection and properties of alternative forming fluids for TRISO fuel kernel production

    Baker, M.P. [Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); King, J.C., E-mail: kingjc@mines.edu [Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); Gorman, B.P. [Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); Marshall, D.W. [Idaho National Laboratory, 2525 N. Fremont Avenue, P.O. Box 1625, Idaho Falls, ID 83415 (United States)

    2013-01-15

    Highlights: Black-Right-Pointing-Pointer Forming fluid selection criteria developed for TRISO kernel production. Black-Right-Pointing-Pointer Ten candidates selected for further study. Black-Right-Pointing-Pointer Density, viscosity, and surface tension measured for first time. Black-Right-Pointing-Pointer Settling velocity and heat transfer rates calculated. Black-Right-Pointing-Pointer Three fluids recommended for kernel production testing. - Abstract: Current Very High Temperature Reactor (VHTR) designs incorporate TRi-structural ISOtropic (TRISO) fuel, which consists of a spherical fissile fuel kernel surrounded by layers of pyrolytic carbon and silicon carbide. An internal sol-gel process forms the fuel kernel using wet chemistry to produce uranium oxyhydroxide gel spheres by dropping a cold precursor solution into a hot column of trichloroethylene (TCE). Over time, gelation byproducts inhibit complete gelation, and the TCE must be purified or discarded. The resulting TCE waste stream contains both radioactive and hazardous materials and is thus considered a mixed hazardous waste. Changing the forming fluid to a non-hazardous alternative could greatly improve the economics of TRISO fuel kernel production. Selection criteria for a replacement forming fluid narrowed a list of {approx}10,800 chemicals to yield ten potential replacement forming fluids: 1-bromododecane, 1-bromotetradecane, 1-bromoundecane, 1-chlorooctadecane, 1-chlorotetradecane, 1-iododecane, 1-iodododecane, 1-iodohexadecane, 1-iodooctadecane, and squalane. The density, viscosity, and surface tension for each potential replacement forming fluid were measured as a function of temperature between 25 Degree-Sign C and 80 Degree-Sign C. Calculated settling velocities and heat transfer rates give an overall column height approximation. 1-bromotetradecane, 1-chlorooctadecane, and 1-iodododecane show the greatest promise as replacements, and future tests will verify their ability to form satisfactory

  11. Systematic metabolic engineering of Methylomicrobium alcaliphilum 20Z for 2,3-butanediol production from methane.

    Nguyen, Anh Duc; Hwang, In Yeub; Lee, Ok Kyung; Kim, Donghyuk; Kalyuzhnaya, Marina G; Mariyana, Rina; Hadiyati, Susila; Kim, Min Sik; Lee, Eun Yeol

    2018-04-16

    Methane is considered a next-generation feedstock, and methanotrophic cell-based biorefinery is attractive for production of a variety of high-value compounds from methane. In this work, we have metabolically engineered Methylomicrobium alcaliphilum 20Z for 2,3-butanediol (2,3-BDO) production from methane. The engineered strain 20Z/pBudK.p, harboring the 2,3-BDO synthesis gene cluster (budABC) from Klebsiella pneumoniae, accumulated 2,3-BDO in methane-fed shake flask cultures with a titer of 35.66 mg/L. Expression of the most efficient gene cluster was optimized using selection of promoters, translation initiation rates (TIR), and the combination of 2,3-BDO synthesis genes from different sources. A higher 2,3-BDO titer of 57.7 mg/L was measured in the 20Z/pNBM-Re strain with budA of K. pneumoniae and budB of Bacillus subtilis under the control of the Tac promoter. The genome-scale metabolic network reconstruction of M. alcaliphilum 20Z enabled in silico gene knockout predictions using an evolutionary programming method to couple growth and 2,3-BDO production. The ldh, ack, and mdh genes in M. alcaliphilum 20Z were identified as potential knockout targets. Pursuing these targets, a triple-mutant strain ∆ldh ∆ack ∆mdh was constructed, resulting in a further increase of the 2,3-BDO titer to 68.8 mg/L. The productivity of this optimized strain was then tested in a fed-batch stirred tank bioreactor, where final product concentrations of up to 86.2 mg/L with a yield of 0.0318 g-(2,3-BDO) /g-CH 4 were obtained under O 2 -limited conditions. This study first demonstrates the strategy of in silico simulation-guided metabolic engineering and represents a proof-of-concept for the production of value-added compounds using systematic approaches from engineered methanotrophs. Copyright © 2018 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  12. (13)C-metabolic flux analysis in S-adenosyl-L-methionine production by Saccharomyces cerevisiae.

    Hayakawa, Kenshi; Kajihata, Shuichi; Matsuda, Fumio; Shimizu, Hiroshi

    2015-11-01

    S-Adenosyl-L-methionine (SAM) is a major biological methyl group donor, and is used as a nutritional supplement and prescription drug. Yeast is used for the industrial production of SAM owing to its high intracellular SAM concentrations. To determine the regulation mechanisms responsible for such high SAM production, (13)C-metabolic flux analysis ((13)C-MFA) was conducted to compare the flux distributions in the central metabolism between Kyokai no. 6 (high SAM-producing) and S288C (control) strains. (13)C-MFA showed that the levels of tricarboxylic acid (TCA) cycle flux in SAM-overproducing strain were considerably increased compared to those in the S228C strain. Analysis of ATP balance also showed that a larger amount of excess ATP was produced in the Kyokai 6 strain because of increased oxidative phosphorylation. These results suggest that high SAM production in Kyokai 6 strains could be attributed to enhanced ATP regeneration with high TCA cycle fluxes and respiration activity. Thus, maintaining high respiration efficiency during cultivation is important for improving SAM production. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  13. Metabolic pathway analysis and kinetic studies for production of nattokinase in Bacillus subtilis.

    Unrean, Pornkamol; Nguyen, Nhung H A

    2013-01-01

    We have constructed a reaction network model of Bacillus subtilis. The model was analyzed using a pathway analysis tool called elementary mode analysis (EMA). The analysis tool was used to study the network capabilities and the possible effects of altered culturing conditions on the production of a fibrinolytic enzyme, nattokinase (NK) by B. subtilis. Based on all existing metabolic pathways, the maximum theoretical yield for NK synthesis in B. subtilis under different substrates and oxygen availability was predicted and the optimal culturing condition for NK production was identified. To confirm model predictions, experiments were conducted by testing these culture conditions for their influence on NK activity. The optimal culturing conditions were then applied to batch fermentation, resulting in high NK activity. The EMA approach was also applied for engineering B. subtilis metabolism towards the most efficient pathway for NK synthesis by identifying target genes for deletion and overexpression that enable the cell to produce NK at the maximum theoretical yield. The consistency between experiments and model predictions proves the feasibility of EMA being used to rationally design culture conditions and genetic manipulations for the efficient production of desired products.

  14. Combination of traditional mutation and metabolic engineering to enhance ansamitocin P-3 production in Actinosynnema pretiosum.

    Du, Zhi-Qiang; Zhang, Yuan; Qian, Zhi-Gang; Xiao, Han; Zhong, Jian-Jiang

    2017-12-01

    Ansamitocin P-3 (AP-3) is a maytansinoid with its most compelling antitumor activity, however, the low production titer of AP-3 greatly restricts its wide commercial application. In this work, a combinatorial approach including random mutation and metabolic engineering was conducted to enhance AP-3 biosynthesis in Actinosynnema pretiosum. First, a mutant strain M was isolated by N-methyl-N'-nitro-N-nitrosoguanidine mutation, which could produce AP-3 almost threefold that of wild type (WT) in 48 deep-well plates. Then, by overexpressing key biosynthetic genes asmUdpg and asm13-17 in the M strain, a further 60% increase of AP-3 production in 250-ml shake flasks was achieved in the engineered strain M-asmUdpg:asm13-17 compared to the M strain, and its maximum AP-3 production reached 582.7 mg/L, which is the highest as ever reported. Both the gene transcription levels and intracellular intermediate concentrations in AP-3 biosynthesis pathway were significantly increased in the M and M-asmUdpg:asm13-17 during fermentation compared to the WT. The good fermentation performance of the engineered strain was also confirmed in a lab-scale bioreactor. This work demonstrated that combination of random mutation and metabolic engineering could promote AP-3 biosynthesis and might be helpful for increasing the production of other industrially important secondary metabolites. © 2017 Wiley Periodicals, Inc.

  15. Spatial distribution of crystalline corrosion products formed during corrosion of stainless steel in concrete

    Serdar, Marijana

    2015-05-01

    © 2015 Elsevier Ltd All rights reserved. The mineralogy and spatial distribution of nano-crystalline corrosion products that form in the steel/concrete interface were characterized using synchrotron X-ray micro-diffraction (μ-XRD). Two types of low-nickel high-chromium reinforcing steels embedded into mortar and exposed to NaCl solution were investigated. Corrosion in the samples was confirmed by electrochemical impedance spectroscopy (EIS). μ-XRD revealed that goethite (α-FeOOH) and akaganeite (β-FeOOH) are the main iron oxide-hydroxides formed during the chloride-induced corrosion of stainless steel in concrete. Goethite is formed closer to the surface of the steel due to the presence of chromium in the steel, while akaganeite is formed further away from the surface due to the presence of chloride ions. Detailed microstructural analysis is shown and discussed on one sample of each type of steel.

  16. Spatial distribution of crystalline corrosion products formed during corrosion of stainless steel in concrete

    Serdar, Marijana; Meral, Cagla; Kunz, Martin; Bjegovic, Dubravka; Wenk, Hans-Rudolf; Monteiro, Paulo J.M.

    2015-01-01

    © 2015 Elsevier Ltd All rights reserved. The mineralogy and spatial distribution of nano-crystalline corrosion products that form in the steel/concrete interface were characterized using synchrotron X-ray micro-diffraction (μ-XRD). Two types of low-nickel high-chromium reinforcing steels embedded into mortar and exposed to NaCl solution were investigated. Corrosion in the samples was confirmed by electrochemical impedance spectroscopy (EIS). μ-XRD revealed that goethite (α-FeOOH) and akaganeite (β-FeOOH) are the main iron oxide-hydroxides formed during the chloride-induced corrosion of stainless steel in concrete. Goethite is formed closer to the surface of the steel due to the presence of chromium in the steel, while akaganeite is formed further away from the surface due to the presence of chloride ions. Detailed microstructural analysis is shown and discussed on one sample of each type of steel.

  17. Metabolic engineering of strains: from industrial-scale to lab-scale chemical production.

    Sun, Jie; Alper, Hal S

    2015-03-01

    A plethora of successful metabolic engineering case studies have been published over the past several decades. Here, we highlight a collection of microbially produced chemicals using a historical framework, starting with titers ranging from industrial scale (more than 50 g/L), to medium-scale (5-50 g/L), and lab-scale (0-5 g/L). Although engineered Escherichia coli and Saccharomyces cerevisiae emerge as prominent hosts in the literature as a result of well-developed genetic engineering tools, several novel native-producing strains are gaining attention. This review catalogs the current progress of metabolic engineering towards production of compounds such as acids, alcohols, amino acids, natural organic compounds, and others.

  18. Advances in metabolic engineering in the microbial production of fuels and chemicals from C1 gas.

    Humphreys, Christopher M; Minton, Nigel P

    2018-04-01

    The future sustainable production of chemicals and fuels from non-petrochemical sources, while at the same time reducing greenhouse gas (GHG) emissions, represent two of society's greatest challenges. Microbial chassis able to grow on waste carbon monoxide (CO) and carbon dioxide (CO 2 ) can provide solutions to both. Ranging from the anaerobic acetogens, through the aerobic chemoautotrophs to the photoautotrophic cyanobacteria, they are able to convert C1 gases into a range of chemicals and fuels which may be enhanced and extended through appropriate metabolic engineering. The necessary improvements will be facilitated by the increasingly sophisticated gene tools that are beginning to emerge as part of the Synthetic Biology revolution. These tools, in combination with more accurate metabolic and genome scale models, will enable C1 chassis to deliver their full potential. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  19. Spatial distribution of crystalline corrosion products formed during corrosion of stainless steel in concrete

    Serdar, Marijana [Department of Materials, Faculty of Civil Engineering, University of Zagreb, 10000 Zagreb (Croatia); Meral, Cagla [Middle East Technical University, Department of Civil Engineering, Ankara (Turkey); Kunz, Martin [Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720 (United States); Bjegovic, Dubravka [Department of Materials, Faculty of Civil Engineering, University of Zagreb, 10000 Zagreb (Croatia); Wenk, Hans-Rudolf [Department of Earth and Planetary Science, University of California, Berkeley, CA 94720 (United States); Monteiro, Paulo J.M., E-mail: monteiro@ce.berkeley.edu [Department of Civil and Environmental Engineering, University of California, Berkeley, CA 94720 (United States)

    2015-05-15

    The mineralogy and spatial distribution of nano-crystalline corrosion products that form in the steel/concrete interface were characterized using synchrotron X-ray micro-diffraction (μ-XRD). Two types of low-nickel high-chromium reinforcing steels embedded into mortar and exposed to NaCl solution were investigated. Corrosion in the samples was confirmed by electrochemical impedance spectroscopy (EIS). μ-XRD revealed that goethite (α-FeOOH) and akaganeite (β-FeOOH) are the main iron oxide–hydroxides formed during the chloride-induced corrosion of stainless steel in concrete. Goethite is formed closer to the surface of the steel due to the presence of chromium in the steel, while akaganeite is formed further away from the surface due to the presence of chloride ions. Detailed microstructural analysis is shown and discussed on one sample of each type of steel. - Highlights: • Synchrotron micro-diffraction used to map the distribution of crystalline phases. • Goethite and akaganeite are the main corrosion products during chloride induced corrosion in mortar. • Layers of goethite and akaganeite are negatively correlated. • EDS showed Cr present in corrosion products identified by SEM.

  20. TEM characterization of corrosion products formed on a SS-15ZR alloy

    Luo, J. S.; Abraham, D. P.

    2000-01-01

    The corrosion products formed on a stainless steel-15Zr (SS-15Zr) alloy have been characterized by transmission electron microscopy (TEM) and energy dispersive x-ray spectroscopy (EDS). Examination of alloy particles that were immersed in 90 C deionized water for two years revealed that different corrosion products were formed on the stainless steel and intermetallic phases. Two corrosion products were identified on an austenite particle: trevorite (NiFe 2 O 4 ) in the layer close to the metal and maghemite (Fe 2 O 3 ) in the outer layer. The corrosion layer formed on the intermetallic was uniform, adherent, and amorphous. The EDS analysis indicated that the layer was enriched in zirconium when compared with the intermetallic composition. High-resolution TEM images of the intermetallic-corrosion layer interface show an interlocking metal-oxide interface which may explain the relatively strong adherence of the corrosion layer to the intermetallic surface. These results will be used to evaluate corrosion mechanisms and predict long-term corrosion behavior of the alloy waste form

  1. Spatial distribution of crystalline corrosion products formed during corrosion of stainless steel in concrete

    Serdar, Marijana; Meral, Cagla; Kunz, Martin; Bjegovic, Dubravka; Wenk, Hans-Rudolf; Monteiro, Paulo J.M.

    2015-01-01

    The mineralogy and spatial distribution of nano-crystalline corrosion products that form in the steel/concrete interface were characterized using synchrotron X-ray micro-diffraction (μ-XRD). Two types of low-nickel high-chromium reinforcing steels embedded into mortar and exposed to NaCl solution were investigated. Corrosion in the samples was confirmed by electrochemical impedance spectroscopy (EIS). μ-XRD revealed that goethite (α-FeOOH) and akaganeite (β-FeOOH) are the main iron oxide–hydroxides formed during the chloride-induced corrosion of stainless steel in concrete. Goethite is formed closer to the surface of the steel due to the presence of chromium in the steel, while akaganeite is formed further away from the surface due to the presence of chloride ions. Detailed microstructural analysis is shown and discussed on one sample of each type of steel. - Highlights: • Synchrotron micro-diffraction used to map the distribution of crystalline phases. • Goethite and akaganeite are the main corrosion products during chloride induced corrosion in mortar. • Layers of goethite and akaganeite are negatively correlated. • EDS showed Cr present in corrosion products identified by SEM

  2. The production of advanced glass ceramic HLW forms using cold crucible induction melter

    Rutledge, V.J.; Maio, V.

    2013-01-01

    Cold Crucible Induction Melters (CCIM) will favorably change how High-Level radioactive Waste (from nuclear fuel recovery) is treated in a near future. Unlike the existing Joule-Heated Melters (JHM) currently in operation for the glass-based immobilization of High-Level Waste (HLW), CCIM offers unique material features that will increase melt temperatures, increase throughput, increase mixing, increase loading in the waste form, lower melter foot prints, eliminate melter corrosion and lower costs. These features not only enhance the technology for producing HLW forms, but also provide advantageous attributes to the waste form by allowing more durable alternatives to glass. It is concluded that glass ceramic waste forms that are tailored to immobilize fission products of HLW can be can be made from the HLW processed with the CCIM. The advantageous higher temperatures reached with the CCIM and unachievable with JHM allows the lanthanides, alkali, alkaline earths, and molybdenum to dissolve into a molten glass. Upon controlled cooling they go into targeted crystalline phases to form a glass ceramic waste form with higher waste loadings than achievable with borosilicate glass waste forms. Natural cooling proves to be too fast for the formation of all targeted crystalline phases

  3. A comparison of the development and metabolic activity of mycorrhizas formed by arbuscular mycorrhizal fungi from different genera on two tropical forage legumes

    Boddington, C.L.; Dodd, J.C.

    1998-01-01

    Two glasshouse experiments were done to assess the development and metabolic activity of mycorrhizas formed by isolates of arbuscular mycorrhizal fungi (AMF) from three different genera, Acaulospora, Gigaspora and Glomus on either Pueraria phaseoloides L. or Desmodium ovalifolium L. plants. The

  4. Multi-omic profiling of EPO producing Chinese hamster ovary cell panel reveals metabolic adaptation to heterologous protein production

    Ley, Daniel; Kazemi Seresht, Ali; Engmark, Mikael

    Heterologous protein production in CHO cells imposes a burden on the host cell metabolism and impact cellular physiology on a global scale. In this work, a multi-omics approach was applied to characterize the physiological impact of erythropoietin production, and discover production bottlenecks, ...

  5. Lactate in the brain: from metabolic end-product to signalling molecule

    Magistretti, Pierre J.

    2018-03-08

    Lactate in the brain has long been associated with ischaemia; however, more recent evidence shows that it can be found there under physiological conditions. In the brain, lactate is formed predominantly in astrocytes from glucose or glycogen in response to neuronal activity signals. Thus, neurons and astrocytes show tight metabolic coupling. Lactate is transferred from astrocytes to neurons to match the neuronal energetic needs, and to provide signals that modulate neuronal functions, including excitability, plasticity and memory consolidation. In addition, lactate affects several homeostatic functions. Overall, lactate ensures adequate energy supply, modulates neuronal excitability levels and regulates adaptive functions in order to set the \\'homeostatic tone\\' of the nervous system.

  6. Advances in metabolic engineering of yeast Saccharomyces cerevisiae for production of chemicals.

    Borodina, Irina; Nielsen, Jens

    2014-05-01

    Yeast Saccharomyces cerevisiae is an important industrial host for production of enzymes, pharmaceutical and nutraceutical ingredients and recently also commodity chemicals and biofuels. Here, we review the advances in modeling and synthetic biology tools and how these tools can speed up the development of yeast cell factories. We also present an overview of metabolic engineering strategies for developing yeast strains for production of polymer monomers: lactic, succinic, and cis,cis-muconic acids. S. cerevisiae has already firmly established itself as a cell factory in industrial biotechnology and the advances in yeast strain engineering will stimulate development of novel yeast-based processes for chemicals production. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Advances in metabolic engineering of yeast Saccharomyces cerevisiae for production of chemicals

    Borodina, Irina; Nielsen, Jens

    2014-01-01

    Yeast Saccharomyces cerevisiae is an important industrial host for production of enzymes, pharmaceutical and nutraceutical ingredients and recently also commodity chemicals and biofuels. Here, we review the advances in modeling and synthetic biology tools and how these tools can speed up the deve......Yeast Saccharomyces cerevisiae is an important industrial host for production of enzymes, pharmaceutical and nutraceutical ingredients and recently also commodity chemicals and biofuels. Here, we review the advances in modeling and synthetic biology tools and how these tools can speed up...... the development of yeast cell factories. We also present an overview of metabolic engineering strategies for developing yeast strains for production of polymer monomers: lactic, succinic, and cis,cis-muconic acids. S. cerevisiae has already firmly established itself as a cell factory in industrial biotechnology...

  8. Improving polyglucan production in cyanobacteria and microalgae via cultivation design and metabolic engineering.

    Aikawa, Shimpei; Ho, Shih-Hsin; Nakanishi, Akihito; Chang, Jo-Shu; Hasunuma, Tomohisa; Kondo, Akihiko

    2015-06-01

    Photosynthetic microorganisms, such as cyanobacteria and microalgae, are currently being investigated as alternative biomass resources for bioethanol production, owing to their benefits, including high-photosynthetic activity and whole-year cultivation without utilization of arable land. Polyglucans comprise the major carbohydrate content of these organisms. Polyglucans can be utilized as a carbon source for microbial fermentation. Although polyglucan production has so far been promoted by nutrient limitation, it must be further enhanced to accommodate market demand. This review focuses on the recent progress in the production of α-polyglucans such asglycogen and starch in cyanobacteria and green microalgae via cultivation design, including modifying the nutrient supply and replacing the growth medium. The control and manipulation of polyglucan metabolism necessitates the elucidation of the polyglucan production mechanism. We reviewed gene expression and metabolite accumulation profiles of cyanobacteria and green microalgae during nutrient limitation-stimulated α-polyglucan accumulation. We also focus on the enhancement in cyanobacterial glycogen production via the genetic engineering of glycolysis, CO2 concentration mechanism, and photosynthetic light-harvesting protein based on the polyglucan accumulation mechanism. The combined strategies of cultivation design and genetic engineering should be considered for further enhancement of polyglucan productivity for bioethanol production. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  9. Dropwise additive manufacturing of pharmaceutical products for melt-based dosage forms.

    Içten, Elçin; Giridhar, Arun; Taylor, Lynne S; Nagy, Zoltan K; Reklaitis, Gintaras V

    2015-05-01

    The US Food and Drug Administration introduced the quality by design approach and process analytical technology guidance to encourage innovation and efficiency in pharmaceutical development, manufacturing, and quality assurance. As part of this renewed emphasis on the improvement of manufacturing, the pharmaceutical industry has begun to develop more efficient production processes with more intensive use of online measurement and sensing, real-time quality control, and process control tools. Here, we present dropwise additive manufacturing of pharmaceutical products (DAMPP) as an alternative to conventional pharmaceutical manufacturing methods. This mini-manufacturing process for the production of pharmaceuticals utilizes drop on demand printing technology for automated and controlled deposition of melt-based formulations onto edible substrates. The advantages of drop-on-demand technology, including reproducible production of small droplets, adjustable drop sizing, high placement accuracy, and flexible use of different formulations, enable production of individualized dosing even for low-dose and high-potency drugs. In this work, DAMPP is used to produce solid oral dosage forms from hot melts of an active pharmaceutical ingredient and a polymer. The dosage forms are analyzed to show the reproducibility of dosing and the dissolution behavior of different formulations. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

  10. Identification and in vitro cytotoxicity of ochratoxin A degradation products formed during coffee roasting.

    Cramer, Benedikt; Königs, Maika; Humpf, Hans-Ulrich

    2008-07-23

    The mycotoxin ochratoxin A is degraded by up to 90% during coffee roasting. In order to investigate this degradation, model heating experiments with ochratoxin A were carried out, and the reaction products were analyzed by HPLC-DAD and HPLC-MS/MS. Two ochratoxin A degradation products were identified, and their structure and absolute configuration were determined. As degradation reactions, the isomerization to 14-(R)-ochratoxin A and the decarboxylation to 14-decarboxy-ochratoxin A were identified. Subsequently, an analytical method for the determination of these compounds in roasted coffee was developed. Quantification was carried out by HPLC-MS/MS and the use of stable isotope dilution analysis. By using this method for the analysis of 15 coffee samples from the German market, it could be shown that, during coffee roasting, the ochratoxin A diastereomer 14-(R)-ochratoxin A was formed in amounts of up to 25.6% relative to ochratoxin A. The decarboxylation product was formed only in traces. For toxicity evaluations, first preliminary cell culture assays were performed with the two new substances. Both degradation products exhibited higher IC50 values and caused apoptotic effects with higher concentrations than ochratoxin A in cultured human kidney epithelial cells. Thus, these cell culture data suggest that the degradation products are less cytotoxic than ochratoxin A.

  11. Processing of spent pickling liquor formed during treatment of titanium products

    Bykovsky, N. A.; Rahman, P. A.; Puchkova, L. N.; Fanakova, N. N.

    2017-10-01

    The article presents the research findings on processing of spent acid pickling liquor (SAPL) formed during etching of titanium products. The processing includes neutralizing the SAPL with alkali, filtering, drying and calcining the titanium hydroxide precipitate as well as electrochemical processing of the filtrate in an ion-exchange membrane cell. The proposed SAPL processing procedure allows obtaining titanium dioxide, sodium hydroxide and a mixture of acids. Titanium dioxide can be used in paint-and-varnish industry. The alkali can be used in neutralizing the SAPL. A mixture of acids is suitable for use in etching process of titanium products.

  12. Metabolic engineering of Pseudomonas fluorescens for the production of vanillin from ferulic acid.

    Di Gioia, Diana; Luziatelli, Francesca; Negroni, Andrea; Ficca, Anna Grazia; Fava, Fabio; Ruzzi, Maurizio

    2011-12-20

    Vanillin is one of the most important flavors in the food industry and there is great interest in its production through biotechnological processes starting from natural substrates such as ferulic acid. Among bacteria, recombinant Escherichia coli strains are the most efficient vanillin producers, whereas Pseudomonas spp. strains, although possessing a broader metabolic versatility, rapidly metabolize various phenolic compounds including vanillin. In order to develop a robust Pseudomonas strain that can produce vanillin in high yields and at high productivity, the vanillin dehydrogenase (vdh)-encoding gene of Pseudomonas fluorescens BF13 strain was inactivated via targeted mutagenesis. The results demonstrated that engineered derivatives of strain BF13 accumulate vanillin if inactivation of vdh is associated with concurrent expression of structural genes for feruloyl-CoA synthetase (fcs) and hydratase/aldolase (ech) from a low-copy plasmid. The conversion of ferulic acid to vanillin was enhanced by optimization of growth conditions, growth phase and parameters of the bioconversion process. The developed strain produced up to 8.41 mM vanillin, which is the highest final titer of vanillin produced by a Pseudomonas strain to date and opens new perspectives in the use of bacterial biocatalysts for biotechnological production of vanillin from agro-industrial wastes which contain ferulic acid. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. Unraveling and engineering the production of 23,24-bisnorcholenic steroids in sterol metabolism.

    Xu, Li-Qin; Liu, Yong-Jun; Yao, Kang; Liu, Hao-Hao; Tao, Xin-Yi; Wang, Feng-Qing; Wei, Dong-Zhi

    2016-02-22

    The catabolism of sterols in mycobacteria is highly important due to its close relevance in the pathogenesis of pathogenic strains and the biotechnological applications of nonpathogenic strains for steroid synthesis. However, some key metabolic steps remain unknown. In this study, the hsd4A gene from Mycobacterium neoaurum ATCC 25795 was investigated. The encoded protein, Hsd4A, was characterized as a dual-function enzyme, with both 17β-hydroxysteroid dehydrogenase and β-hydroxyacyl-CoA dehydrogenase activities in vitro. Using a kshAs-null strain of M. neoaurum ATCC 25795 (NwIB-XII) as a model, Hsd4A was further confirmed to exert dual-function in sterol catabolism in vivo. The deletion of hsd4A in NwIB-XII resulted in the production of 23,24-bisnorcholenic steroids (HBCs), indicating that hsd4A plays a key role in sterol side-chain degradation. Therefore, two competing pathways, the AD and HBC pathways, were proposed for the side-chain degradation. The proposed HBC pathway has great value in illustrating the production mechanism of HBCs in sterol catabolism and in developing HBCs producing strains for industrial application via metabolic engineering. Through the combined modification of hsd4A and other genes, three HBCs producing strains were constructed that resulted in promising productivities of 0.127, 0.109 and 0.074 g/l/h, respectively.

  14. Metabolic engineering of Escherichia coli for biotechnological production of high-value organic acids and alcohols

    Yu, Chao; Cao, Yujin; Zou, Huibin; Xian, Mo [Chinese Academy of Sciences, Qingdao (China). Key Lab. of Biofuels

    2011-02-15

    Confronted with the gradual and inescapable exhaustion of the earth's fossil energy resources, the bio-based process to produce platform chemicals from renewable carbohydrates is attracting growing interest. Escherichia coli has been chosen as a workhouse for the production of many valuable chemicals due to its clear genetic background, convenient to be genetically modified and good growth properties with low nutrient requirements. Rational strain development of E. coli achieved by metabolic engineering strategies has provided new processes for efficiently biotechnological production of various high-value chemical building blocks. Compared to previous reviews, this review focuses on recent advances in metabolic engineering of the industrial model bacteria E. coli that lead to efficient recombinant biocatalysts for the production of high-value organic acids like succinic acid, lactic acid, 3-hydroxypropanoic acid and glucaric acid as well as alcohols like 1,3-propanediol, xylitol, mannitol, and glycerol with the discussion of the future research in this area. Besides, this review also discusses several platform chemicals, including fumaric acid, aspartic acid, glutamic acid, sorbitol, itaconic acid, and 2,5-furan dicarboxylic acid, which have not been produced by E. coli until now. (orig.)

  15. Probiotics and Probiotic Metabolic Product Improved Intestinal Function and Ameliorated LPS-Induced Injury in Rats.

    Deng, Bo; Wu, Jie; Li, Xiaohui; Men, Xiaoming; Xu, Ziwei

    2017-11-01

    In the present study, we sought to determine the effects of Bacillus subtilis (BAS) and Bacillus licheniformis (BAL) in rats after lipopolysaccharide (LPS)-induced acute intestinal inflammation. We also determined whether the B. subtilis metabolic product (BASM) is as effective as the live-cell probiotic. 60 male SD rats were randomly assigned to five groups and administered a diet containing 0.05% B. licheniformis (BAL group), 0.05% B. subtilis (BAS group), 0.5% B. subtilis metabolic product (BASM group), or a basic diet (PC group and NC group) for 40 days. On day 40, BAL, BAS, BASM, and NC groups were injected with 4 mg/kg body weight LPS. 4 h later, all rats were anesthetized and sacrificed. The results showed that the administration of B. licheniformis and B. subtilis improved intestinal function as evidenced by histology, increased enzyme activity, and mucosal thickness. They also increased the number of intraepithelial lymphocytes and decreased mucosal myeloperoxidase activity and plasma TNF-α. In addition, the cecal content of B. subtilis-treated rats had significantly increased microbial diversity, decreased numbers of Firmicutes, and increased numbers of Bacteroidetes as compared to rats fed basic diets. Similar to BAS group, the cecal content of B. licheniformis-treated rats decreased the number of Firmicutes. Administration of B. subtilis metabolic product had similar effects on intestinal function, inflammation response, and microbial diversity as B. subtilis but these effects were attenuated. In conclusion, administration of probiotic strains B. licheniformis or B. subtilis improved intestinal function, ameliorated the inflammation response, and modulated microflora after LPS-induced acute inflammation in rats. Non-living cells also exerted probiotic properties but live cells tended to function better.

  16. Collection and application of by-product formed in e-b flue gas treatment process

    Chmielewski, A.G.; Tyminski, B.; Zakrzewska-Trznadel, G.; Tokunaga, O.; Machi, S.

    1998-01-01

    In the e-b process SO 2 and NO x are converted into ammonium sulphate and ammonium nitrate, which condenses from gas phase in the form of submicrone particles. These salts are a valuable fertilizer and should be removed from cleaned gas. Bag filter, ESP and wet gravel bed filter were applied for collecting of salt particles in pilot plant facilities. Up to now ESP is considered to be the best filtration method of aerosols formed after irradiation of flue gas. Collected salts after granulation may be used as a fertilizer enriching soil in nitrogen and sulphur or as a component of mixed fertilizer. Analysis of by-products from different e-b pilot plants confirms that it does not contain any harmful substances like heavy metals and fulfill all standards for commercial fertilizers. Also field experiments show that the by-products have the same properties as a commercial fertilizer

  17. Metabolic flux profiling of MDCK cells during growth and canine adenovirus vector production

    Nuno Carinhas; Daniel A. M. Pais; Alexey Koshkin; Paulo Fernandes; Ana S. Coroadinha; Manuel J. T. Carrondo; Paula M. Alves; Ana P. Teixeira

    2016-01-01

    Canine adenovirus vector type 2 (CAV2) represents an alternative to human adenovirus vectors for certain gene therapy applications, particularly neurodegenerative diseases. However, more efficient production processes, assisted by a greater understanding of the effect of infection on producer cells, are required. Combining [1,2-13C]glucose and [U-13C]glutamine, we apply for the first time 13C-Metabolic flux analysis (13C-MFA) to study E1-transformed Madin-Darby Canine Kidney (MDCK) cells meta...

  18. A20 modulates lipid metabolism and energy production to promote liver regeneration.

    Scott M Damrauer

    2011-03-01

    Full Text Available Liver regeneration is clinically of major importance in the setting of liver injury, resection or transplantation. We have demonstrated that the NF-κB inhibitory protein A20 significantly improves recovery of liver function and mass following extended liver resection (LR in mice. In this study, we explored the Systems Biology modulated by A20 following extended LR in mice.We performed transcriptional profiling using Affymetrix-Mouse 430.2 arrays on liver mRNA retrieved from recombinant adenovirus A20 (rAd.A20 and rAd.βgalactosidase treated livers, before and 24 hours after 78% LR. A20 overexpression impacted 1595 genes that were enriched for biological processes related to inflammatory and immune responses, cellular proliferation, energy production, oxidoreductase activity, and lipid and fatty acid metabolism. These pathways were modulated by A20 in a manner that favored decreased inflammation, heightened proliferation, and optimized metabolic control and energy production. Promoter analysis identified several transcriptional factors that implemented the effects of A20, including NF-κB, CEBPA, OCT-1, OCT-4 and EGR1. Interactive scale-free network analysis captured the key genes that delivered the specific functions of A20. Most of these genes were affected at basal level and after resection. We validated a number of A20's target genes by real-time PCR, including p21, the mitochondrial solute carriers SLC25a10 and SLC25a13, and the fatty acid metabolism regulator, peroxisome proliferator activated receptor alpha. This resulted in greater energy production in A20-expressing livers following LR, as demonstrated by increased enzymatic activity of cytochrome c oxidase, or mitochondrial complex IV.This Systems Biology-based analysis unravels novel mechanisms supporting the pro-regenerative function of A20 in the liver, by optimizing energy production through improved lipid/fatty acid metabolism, and down-regulated inflammation. These findings

  19. Degradation and metabolism of synthetic plastics and associated products by Pseudomonas sp.: capabilities and challenges.

    Wilkes, R A; Aristilde, L

    2017-09-01

    Synthetic plastics, which are widely present in materials of everyday use, are ubiquitous and slowly-degrading polymers in environmental wastes. Of special interest are the capabilities of microorganisms to accelerate their degradation. Members of the metabolically diverse genus Pseudomonas are of particular interest due to their capabilities to degrade and metabolize synthetic plastics. Pseudomonas species isolated from environmental matrices have been identified to degrade polyethylene, polypropylene, polyvinyl chloride, polystyrene, polyurethane, polyethylene terephthalate, polyethylene succinate, polyethylene glycol and polyvinyl alcohol at varying degrees of efficiency. Here, we present a review of the current knowledge on the factors that control the ability of Pseudomonas sp. to process these different plastic polymers and their by-products. These factors include cell surface attachment within biofilms, catalytic enzymes involved in oxidation or hydrolysis of the plastic polymer, metabolic pathways responsible for uptake and assimilation of plastic fragments and chemical factors that are advantageous or inhibitory to the biodegradation process. We also highlight future research directions required in order to harness fully the capabilities of Pseudomonas sp. in bioremediation strategies towards eliminating plastic wastes. © 2017 The Society for Applied Microbiology.

  20. Biosynthesis and metabolic engineering of palmitoleate production, an important contributor to human health and sustainable industry.

    Wu, Yongmei; Li, Runzhi; Hildebrand, David F

    2012-10-01

    Palmitoleate (cis-Δ9-16:1) shows numerous health benefits such as increased cell membrane fluidity, reduced inflammation, protection of the cardiovascular system, and inhibition of oncogenesis. Plant oils containing this unusual fatty acid can also be sustainable feedstocks for producing industrially important and high-demand 1-octene. Vegetable oils rich in palmitoleate are the ideal candidates for biodiesel production. Several wild plants are known that can synthesize high levels of palmitoleate in seeds. However, low yields and poor agronomic characteristics of these plants limit their commercialization. Metabolic engineering has been developed to create oilseed crops that accumulate high levels of palmitoleate or other unusual fatty acids, and significant advances have been made recently in this field, particularly using the model plant Arabidopsis as the host. The engineered targets for enhancing palmitoleate synthesis include overexpression of Δ9 desaturase from mammals, yeast, fungi, and plants, down-regulating KASII, coexpression of an ACP-Δ9 desaturase in plastids and CoA-Δ9 desaturase in endoplasmic reticulum (ER), and optimizing the metabolic flux into triacylglycerols (TAGs). This review will mainly describe the recent progress towards producing palmitoleate in transgenic plants by metabolic engineering along with our current understanding of palmitoleate biosynthesis and its regulation, as well as highlighting the bottlenecks that require additional investigation by combining lipidomics, transgenics and other "-omics" tools. A brief review of reported health benefits and non-food uses of palmitoleate will also be covered. Copyright © 2012. Published by Elsevier Ltd.

  1. Quality assurance in production and use of special form radioactive material - focal points in BAM approvals

    Rolle, A.; Buhlemann, L.

    2004-01-01

    BAM as the competent authority for approval of special form radioactive material attaches great importance to a detailed audit of the required quality assurance programs for design, manufacture, testing, documentation, use, maintenance and inspection. Applicants have to submit, together with application documentation information on general arrangements for quality assurance, as well as on quality assurance in production and in operation. Fields where BAM has often found deficiencies are leak test methods, weld seam quality and the safety level after use

  2. Diagnostic value of daily fluctuations in the free form of testosterone and cortisol in men with obesity and metabolic syndrome under the age of 50 years

    E. A. Kuznetsova; A. S. Adamchik; N. P. Goncharov; G. V. Katsiya

    2016-01-01

    The important pathogenetic link of obesity and metabolic syndrome (MS) in men are disorders of testosterone production in the testes and adrenal adrenal cortisol production. Тestosterone deficiency and functional hypercortisolism have a mutual influence on each other. Comprehensive assessment of testosterone and cortisol balance and identify their daily variability in the saliva may improve the diagnosis of hormonal disorders in men with obesity and MS. In the present study, by comparative an...

  3. Recent advances in microbial production of fuels and chemicals using tools and strategies of systems metabolic engineering

    Cho, Changhee; Choi, So Young; Luo, Zi Wei

    2015-01-01

    The advent of various systems metabolic engineering tools and strategies has enabled more sophisticated engineering of microorganisms for the production of industrially useful fuels and chemicals. Advances in systems metabolic engineering have been made in overproducing natural chemicals...... and producing novel non-natural chemicals. In this paper, we review the tools and strategies of systems metabolic engineering employed for the development of microorganisms for the production of various industrially useful chemicals belonging to fuels, building block chemicals, and specialty chemicals......, in particular focusing on those reported in the last three years. It was aimed at providing the current landscape of systems metabolic engineering and suggesting directions to address future challenges towards successfully establishing processes for the bio-based production of fuels and chemicals from renewable...

  4. Recent advances in microbial production of fuels and chemicals using tools and strategies of systems metabolic engineering.

    Cho, Changhee; Choi, So Young; Luo, Zi Wei; Lee, Sang Yup

    2015-11-15

    The advent of various systems metabolic engineering tools and strategies has enabled more sophisticated engineering of microorganisms for the production of industrially useful fuels and chemicals. Advances in systems metabolic engineering have been made in overproducing natural chemicals and producing novel non-natural chemicals. In this paper, we review the tools and strategies of systems metabolic engineering employed for the development of microorganisms for the production of various industrially useful chemicals belonging to fuels, building block chemicals, and specialty chemicals, in particular focusing on those reported in the last three years. It was aimed at providing the current landscape of systems metabolic engineering and suggesting directions to address future challenges towards successfully establishing processes for the bio-based production of fuels and chemicals from renewable resources. Copyright © 2014 Elsevier Inc. All rights reserved.

  5. Metabolic engineering of riboflavin production in Ashbya gossypii through pathway optimization.

    Ledesma-Amaro, Rodrigo; Serrano-Amatriain, Cristina; Jiménez, Alberto; Revuelta, José Luis

    2015-10-14

    The industrial production of riboflavin mostly relies on the microbial fermentation of flavinogenic microorganisms and Ashbya gossypii is the main industrial producer of the vitamin. Accordingly, bioengineering strategies aimed at increasing riboflavin production in A. gossypii are highly valuable for industry. We analyze the contribution of all the RIB genes to the production of riboflavin in A. gossypii. Two important metabolic rate-limiting steps that limit the overproduction of riboflavin have been found: first, low mRNA levels of the RIB genes hindered the overproduction of riboflavin; second, the competition of the AMP branch for purinogenic precursors also represents a limitation for riboflavin overproduction. Thus, overexpression of the RIB genes resulted in a significant increase in riboflavin yield. Moreover, both the inactivation and the underexpression of the ADE12 gene, which controls the first step of the AMP branch, also proved to have a positive effect on riboflavin production. Accordingly, a strain that combines both the overexpression of the RIB genes and the underexpression of the ADE12 gene was engineered. This strain produced 523 mg/L of riboflavin (5.4-fold higher than the wild-type), which is the highest titer of riboflavin obtained by metabolic engineering in A. gossypii so far. Riboflavin production in A. gossypii is limited by a low transcription activity of the RIB genes. Flux limitation towards AMP provides committed substrate GTP for riboflavin overproduction without detrimental effects on biomass formation. A multiple-engineered Ashbya strain that produces up to 523 mg/L of riboflavin was generated.

  6. Effects of lactone, ketone, and phenolic compounds on methane production and metabolic intermediates during anaerobic digestion.

    Wikandari, Rachma; Sari, Noor Kartika; A'yun, Qurrotul; Millati, Ria; Cahyanto, Muhammad Nur; Niklasson, Claes; Taherzadeh, Mohammad J

    2015-02-01

    Fruit waste is a potential feedstock for biogas production. However, the presence of fruit flavors that have antimicrobial activity is a challenge for biogas production. Lactones, ketones, and phenolic compounds are among the several groups of fruit flavors that are present in many fruits. This work aimed to investigate the effects of two lactones, i.e., γ-hexalactone and γ-decalactone; two ketones, i.e., furaneol and mesifurane; and two phenolic compounds, i.e., quercetin and epicatechin on anaerobic digestion with a focus on methane production, biogas composition, and metabolic intermediates. Anaerobic digestion was performed in a batch glass digester incubated at 55 °C for 30 days. The flavor compounds were added at concentrations of 0.05, 0.5, and 5 g/L. The results show that the addition of γ-decalactone, quercetin, and epicathechin in the range of 0.5-5 g/L reduced the methane production by 50 % (MIC50). Methane content was reduced by 90 % with the addition of 5 g/L of γ-decalactone, quercetin, and epicathechin. Accumulation of acetic acid, together with an increase in carbon dioxide production, was observed. On the contrary, γ-hexalactone, furaneol, and mesifurane increased the methane production by 83-132 % at a concentration of 5 g/L.

  7. Metabolic engineering of Corynebacterium glutamicum aimed at alternative carbon sources and new products

    Volker Fritz Wendisch

    2012-10-01

    Full Text Available Corynebacterium glutamicum is well known as the amino acid-producing workhorse of fermentation industry, being used for multi-million-ton scale production of glutamate and lysine for more than 60 years. However, it is only recently that extensive research has focused on engineering it beyond the scope of amino acids. Meanwhile, a variety of corynebacterial strains allows access to alternative carbon sources and/or allows production of a wide range of industrially relevant compounds. Some of these efforts set new standards in terms of titers and productivities achieved whereas others represent a proof-of-principle. These achievements manifest the position of C. glutamicum as an important industrial microorganism with capabilities far beyond the traditional amino acid production. In this review we focus on the state of the art of metabolic engineering of C. glutamicum for utilization of alternative carbon sources, (e.g. coming from wastes and unprocessed sources, and construction of C. glutamicum strains for production of new products such as diamines, organic acids and alcohols.

  8. Production of ethanol from thin stillage by metabolically engineered Escherichia coli.

    Gonzalez, Ramon; Campbell, Paul; Wong, Matthew

    2010-03-01

    Thin stillage is a by-product generated in large amounts during the production of ethanol that is rich in carbon sources like glycerol, glucose and maltose. Unfortunately, the fermentation of thin stillage results in a mixture of organic acids and ethanol and minimum utilization of glycerol, the latter a compound that can represent up to 80% of the available substrates in this stream. We report here the efficient production of ethanol from thin stillage by a metabolically engineered strain of Escherichia coli. Simultaneous utilization of glycerol and sugars was achieved by overexpressing either the fermentative or the respiratory glycerol-utilization pathway. However, amplification of the fermentative pathway (encoded by gldA and dhaKLM) led to more efficient consumption of glycerol and promoted the synthesis of reduced products, including ethanol. A previously constructed strain, EH05, containing mutations that prevented the accumulation of competing by-products (i.e. lactate, acetate, and succinate) and overexpressing the fermentative pathway for glycerol utilization [i.e. strain EH05 (pZSKLMgldA)], efficiently converted thin stillage supplemented with only mineral salts to ethanol at yields close to 85% of the theoretical maximum. Ethanol accounted for about 90% (w/w) of the product mixture. These results, along with the comparable performance of strain EH05 (pZSKLMgldA) in 0.5 and 5 l fermenters, indicate a great potential for the adoption of this process by the biofuels industry.

  9. Metabolic network model guided engineering ethylmalonyl-CoA pathway to improve ascomycin production in Streptomyces hygroscopicus var. ascomyceticus.

    Wang, Junhua; Wang, Cheng; Song, Kejing; Wen, Jianping

    2017-10-03

    Ascomycin is a 23-membered polyketide macrolide with high immunosuppressant and antifungal activity. As the lower production in bio-fermentation, global metabolic analysis is required to further explore its biosynthetic network and determine the key limiting steps for rationally engineering. To achieve this goal, an engineering approach guided by a metabolic network model was implemented to better understand ascomycin biosynthesis and improve its production. The metabolic conservation of Streptomyces species was first investigated by comparing the metabolic enzymes of Streptomyces coelicolor A3(2) with those of 31 Streptomyces strains, the results showed that more than 72% of the examined proteins had high sequence similarity with counterparts in every surveyed strain. And it was found that metabolic reactions are more highly conserved than the enzymes themselves because of its lower diversity of metabolic functions than that of genes. The main source of the observed metabolic differences was from the diversity of secondary metabolism. According to the high conservation of primary metabolic reactions in Streptomyces species, the metabolic network model of Streptomyces hygroscopicus var. ascomyceticus was constructed based on the latest reported metabolic model of S. coelicolor A3(2) and validated experimentally. By coupling with flux balance analysis and using minimization of metabolic adjustment algorithm, potential targets for ascomycin overproduction were predicted. Since several of the preferred targets were highly associated with ethylmalonyl-CoA biosynthesis, two target genes hcd (encoding 3-hydroxybutyryl-CoA dehydrogenase) and ccr (encoding crotonyl-CoA carboxylase/reductase) were selected for overexpression in S. hygroscopicus var. ascomyceticus FS35. Both the mutants HA-Hcd and HA-Ccr showed higher ascomycin titer, which was consistent with the model predictions. Furthermore, the combined effects of the two genes were evaluated and the strain HA

  10. Extremely Thermophilic Microorganisms as Metabolic Engineering Platforms for Production of Fuels and Industrial Chemicals

    Benjamin M Zeldes

    2015-11-01

    Full Text Available Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye towards potential technological

  11. Extremely thermophilic microorganisms as metabolic engineering platforms for production of fuels and industrial chemicals

    Zeldes, Benjamin M.; Keller, Matthew W.; Loder, Andrew J.; Straub, Christopher T.; Adams, Michael W. W.; Kelly, Robert M.

    2015-01-01

    Enzymes from extremely thermophilic microorganisms have been of technological interest for some time because of their ability to catalyze reactions of industrial significance at elevated temperatures. Thermophilic enzymes are now routinely produced in recombinant mesophilic hosts for use as discrete biocatalysts. Genome and metagenome sequence data for extreme thermophiles provide useful information for putative biocatalysts for a wide range of biotransformations, albeit involving at most a few enzymatic steps. However, in the past several years, unprecedented progress has been made in establishing molecular genetics tools for extreme thermophiles to the point that the use of these microorganisms as metabolic engineering platforms has become possible. While in its early days, complex metabolic pathways have been altered or engineered into recombinant extreme thermophiles, such that the production of fuels and chemicals at elevated temperatures has become possible. Not only does this expand the thermal range for industrial biotechnology, it also potentially provides biodiverse options for specific biotransformations unique to these microorganisms. The list of extreme thermophiles growing optimally between 70 and 100°C with genetic toolkits currently available includes archaea and bacteria, aerobes and anaerobes, coming from genera such as Caldicellulosiruptor, Sulfolobus, Thermotoga, Thermococcus, and Pyrococcus. These organisms exhibit unusual and potentially useful native metabolic capabilities, including cellulose degradation, metal solubilization, and RuBisCO-free carbon fixation. Those looking to design a thermal bioprocess now have a host of potential candidates to choose from, each with its own advantages and challenges that will influence its appropriateness for specific applications. Here, the issues and opportunities for extremely thermophilic metabolic engineering platforms are considered with an eye toward potential technological advantages for high

  12. Characterization of biofilm-forming cyanobacteria for biomass and lipid production.

    Bruno, L; Di Pippo, F; Antonaroli, S; Gismondi, A; Valentini, C; Albertano, P

    2012-11-01

    This work reports on one of the first attempts to use biofilm-forming cyanobacteria for biomass and lipid production. Three isolates of filamentous cyanobacteria were obtained from biofilms at different Italian sites and characterized by a polyphasic approach, involving microscopic observations, ecology and genetic diversity (studying the 16S rRNA gene). The isolates were grown in batch systems and in a semi-continuous flow incubator, specifically designed for biofilms development. Culture system affected biomass and lipid production, but did not influence the fatty acid profile. The composition of fatty acids was mainly palmitic acid (>50%) and less amounts of other saturated and monounsaturated fatty acids. Only two isolates contained two polyunsaturated fatty acids. Data obtained from the flow-lane incubator system would support a more economical and sustainable use of the benthic micro-organisms for biomass production. The produced lipids contained fatty acids suitable for a high-quality biodiesel production, showing high proportions of saturated and monounsaturated fatty acids. Data seem promising when taking into account the savings in cost and time derived from easy procedures for biomass harvesting, especially when being able to obtain the co-production of other valuable by-products. © 2012 The Authors Journal of Applied Microbiology © 2012 The Society for Applied Microbiology.

  13. Mitochondrial energy metabolism changes during aging-mouse cranial nerve cells treated with various doses and forms of Fructus schizandrae

    Hongyan Guo; Jinhe Li

    2008-01-01

    BACKGROUND: During the cellular aging process, the number of mitochondria, generation of adenosine triphosphate (ATP), activity of respiratory chain enzyme complex 1 and 4, and oxidation decrease. OBJECTIVE: To observe the effects of aqueous and spirituous extract, as well as polysaccharides from Fructus schizandrae (Magnolia Vine) on energy metabolism and mitochondrial anti-oxidation in cranial nerve cells of a D-gal-induccd aging mouse model.DESIGN, TIME AND SETTING: A randomized, controlled, animal study. The experiment was conducted at the Department of Biochemistry, Qiqihar Medical College between March and July 2006.MATERIALS: Fifty healthy, Kunming mice of both sexes, aged 2 3 months old and weighing 18-22 g, were used for the present study. Fructus schizandrae was purchased from the Medical College of Jiamusi University. Aqueous extracts, spirituous extracts, and polysaccharides from Fructus schizandrae were prepared. D-galactose (D-gal) is a product of the Second Reagent Factory, Shanghai City, China. Mn-superoxide dismutase (Mn-SOD) kit, malonaldehyde (MDA) kit, protein quantification kit, and inorganic phosphorus testing kit were purchased from Jian Cheng Bioeng. Co., China.METHODS: Fifty mice were randomly divided into five groups, with 10 mice in each group: young control, aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides. Over a course of 30 days, mice in aging model, aqueous Fructus schizandrae extract, spirituous Fructus schizandrae extract, and Fructus schizandrae polysaccharides groups were injected subcutaneously with D-gal (100 mg/kg) into the nape of the neck daily, and administered intragastrically with an equal volume of sterile, warm water (aging model), aqueous Fructus schizandrae extract (2 g/kg), spirituous Fructus schizandrae extract (2 g/kg), or Fructus schizandrae polysaccharides (0.2 g/kg), respectively. Mice in the young control group were injected into

  14. Feeding Diets Containing Different Forms of Duckweed on Productive Performance and Egg Quality of Ducks

    B. Indarsih

    2012-08-01

    Full Text Available The present experiment was undertaken to study the feeding effect of diets containing different forms of duckweed for local ducks on their productive performance and egg quality or egg yolk pigmentation. A total of 90 birds of 24 wk old ducks were randomly divided into 18 experimental units of 2.0 x 1.0 m2 of cages. The experiment was assigned in a completely randomized design (3 treatments with 6 replicates, 5 birds each. There were 3 dietary treatments, namely P1= ducks fed a complete diet containing 20 % of dried duckweed and given in the form of dry-mash; P2= a complete diet in P1 but it was offered in wet form (slurry; and P3= ducks were offered basal diet in the form of dry-mash and fresh duckweed was offered separately ad libitum. Diets were formulated to have similar nutritional contents. Feed consumption, feed conversion ratio (FCR, and egg yolk pigmentation were measured. The result of the study showed that these three parameters were affected by the feeding different forms of duckweed. Feeding diet with fresh duckweed brought about the best pigmentation than did the dry one. Fresh duckweed offers a promise as a potential feedstuff for ducks and has a good implication in reducing feed processing cost.

  15. Understanding Disorder Within Variation: Production of English Grammatical Forms by English Language Learners.

    Bedore, Lisa M; Peña, Elizabeth D; Anaya, Jissel B; Nieto, Ricardo; Lugo-Neris, Mirza J; Baron, Alisa

    2018-04-05

    This study examines English performance on a set of 11 grammatical forms in Spanish-English bilingual, school-age children in order to understand how item difficulty of grammatical constructions helps correctly classify language impairment (LI) from expected variability in second language acquisition when taking into account linguistic experience and exposure. Three hundred seventy-eight children's scores on the Bilingual English-Spanish Assessment-Middle Extension (Peña, Bedore, Gutiérrez-Clellen, Iglesias, & Goldstein, 2008) morphosyntax cloze task were analyzed by bilingual experience groups (high Spanish experience, balanced English-Spanish experience, high English experience, ability (typically developing [TD] vs. LI), and grammatical form. Classification accuracy was calculated for the forms that best differentiated TD and LI groups. Children with LI scored lower than TD children across all bilingual experience groups. There were differences by grammatical form across bilingual experience and ability groups. Children from high English experience and balanced English-Spanish experience groups could be accurately classified on the basis of all the English grammatical forms tested except for prepositions. For bilinguals with high Spanish experience, it was possible to rule out LI on the basis of grammatical production but not rule in LI. It is possible to accurately identify LI in English language learners once they use English 40% of the time or more. However, for children with high Spanish experience, more information about development and patterns of impairment is needed to positively identify LI.

  16. Effects of chemical form of sodium on the product characteristics of alkali lignin pyrolysis.

    Guo, Da-liang; Yuan, Hong-you; Yin, Xiu-li; Wu, Chuang-zhi; Wu, Shu-bin; Zhou, Zhao-qiu

    2014-01-01

    The effects of Na as organic bound form or as inorganic salts form on the pyrolysis products characteristics of alkali lignin were investigated by using thermogravimetric analyzer coupled with Fourier transform infrared spectrometry (TG-FTIR), tube furnace and thermo-gravimetric analyzer (TGA). Results of TG-FTIR and tube furnace indicated that the two chemical forms Na reduced the releasing peak temperature of CO and phenols leading to the peak temperature of the maximum mass loss rate shifted to low temperature zone. Furthermore, organic bound Na obviously improved the elimination of alkyl substituent leading to the yields of phenol and guaiacol increased, while inorganic Na increased the elimination of phenolic hydroxyl groups promoting the formation of ethers. It was also found the two chemical forms Na had different effects on the gasification reactivity of chars. For inorganic Na, the char conversion decreased with increasing the char forming temperature, while organic bound Na was opposite. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  17. Differential proteomic analysis highlights metabolic strategies associated with balhimycin production in Amycolatopsis balhimycina chemostat cultivations

    Gallo, Giuseppe; Alduina, Rosa; Renzone, Giovanni

    2010-01-01

    Background Proteomics was recently used to reveal enzymes whose expression is associated with the production of the glycopeptide antibiotic balhimycin in Amycolatopsis balhimycina batch cultivations. Combining chemostat fermentation technology, where cells proliferate with constant parameters...... in a highly reproducible steady-state, and differential proteomics, the relationships between physiological status and metabolic pathways during antibiotic producing and non-producing conditions could be highlighted. Results Two minimal defined media, one with low Pi (0.6 mM; LP) and proficient glucose (12 g....../l) concentrations and the other one with high Pi (1.8 mM) and limiting (6 g/l; LG) glucose concentrations, were developed to promote and repress antibiotic production, respectively, in A. balhimycina chemostat cultivations. Applying the same dilution rate (0.03 h-1), both LG and LP chemostat cultivations showed...

  18. Mathematical modeling of unicellular microalgae and cyanobacteria metabolism for biofuel production.

    Baroukh, Caroline; Muñoz-Tamayo, Rafael; Bernard, Olivier; Steyer, Jean-Philippe

    2015-06-01

    The conversion of microalgae lipids and cyanobacteria carbohydrates into biofuels appears to be a promising source of renewable energy. This requires a thorough understanding of their carbon metabolism, supported by mathematical models, in order to optimize biofuel production. However, unlike heterotrophic microorganisms that utilize the same substrate as sources of energy and carbon, photoautotrophic microorganisms require light for energy and CO2 as carbon source. Furthermore, they are submitted to permanent fluctuating light environments due to outdoor cultivation or mixing inducing a flashing effect. Although, modeling these nonstandard organisms is a major challenge for which classical tools are often inadequate, this step remains a prerequisite towards efficient optimization of outdoor biofuel production at an industrial scale. Copyright © 2015 Elsevier Ltd. All rights reserved.

  19. Composition and Metabolic Activities of the Bacterial Community in Shrimp Sauce at the Flavor-Forming Stage of Fermentation As Revealed by Metatranscriptome and 16S rRNA Gene Sequencings.

    Duan, Shan; Hu, Xiaoxi; Li, Mengru; Miao, Jianyin; Du, Jinghe; Wu, Rongli

    2016-03-30

    The bacterial community and the metabolic activities involved at the flavor-forming stage during the fermentation of shrimp sauce were investigated using metatranscriptome and 16S rRNA gene sequencings. Results showed that the abundance of Tetragenococcus was 95.1%. Tetragenococcus halophilus was identified in 520 of 588 transcripts annotated in the Nr database. Activation of the citrate cycle and oxidative phosphorylation, along with the absence of lactate dehydrogenase gene expression, in T. halophilus suggests that T. halophilus probably underwent aerobic metabolism during shrimp sauce fermentation. The metabolism of amino acids, production of peptidase, and degradation of limonene and pinene were very active in T. halophilus. Carnobacterium, Pseudomonas, Escherichia, Staphylococcus, Bacillus, and Clostridium were also metabolically active, although present in very small populations. Enterococcus, Abiotrophia, Streptococcus, and Lactobacillus were detected in metatranscriptome sequencing, but not in 16S rRNA gene sequencing. Many minor taxa showed no gene expression, suggesting that they were in dormant status.

  20. Synthetic protein scaffolds based on peptide motifs and cognate adaptor domains for improving metabolic productivity

    Anselm H.C. Horn

    2015-11-01

    Full Text Available The efficiency of many cellular processes relies on the defined interaction among different proteins within the same metabolic or signaling pathway. Consequently, a spatial colocalization of functionally interacting proteins has frequently emerged during evolution. This concept has been adapted within the synthetic biology community for the purpose of creating artificial scaffolds. A recent advancement of this concept is the use of peptide motifs and their cognate adaptor domains. SH2, SH3, GBD, and PDZ domains have been used most often in research studies to date. The approach has been successfully applied to the synthesis of a variety of target molecules including catechin, D-glucaric acid, H2, hydrochinone, resveratrol, butyrate, gamma-aminobutyric acid, and mevalonate. Increased production levels of up to 77-fold have been observed compared to non-scaffolded systems. A recent extension of this concept is the creation of a covalent linkage between peptide motifs and adaptor domains, which leads to a more stable association of the scaffolded systems and thus bears the potential to further enhance metabolic productivity.

  1. Efficient odd straight medium chain free fatty acid production by metabolically engineered Escherichia coli.

    Wu, Hui; San, Ka-Yiu

    2014-11-01

    Free fatty acids (FFAs) can be used as precursors for the production of biofuels or chemicals. Different composition of FFAs will be useful for further modification of the biofuel/biochemical quality. Microbial biosynthesis of even chain FFAs can be achieved by introducing an acyl-acyl carrier protein thioesterase gene into E. coli. In this study, odd straight medium chain FFAs production was investigated by using metabolic engineered E. coli carrying acyl-ACP thioesterase (TE, Ricinus communis), propionyl-CoA synthase (Salmonella enterica), and β-ketoacyl-acyl carrier protein synthase III (four different sources) with supplement of extracellular propionate. By using these metabolically engineered E. coli, significant quantity of C13 and C15 odd straight-chain FFAs could be produced from glucose and propionate. The highest concentration of total odd straight chain FFAs attained was 1205 mg/L by the strain HWK201 (pXZ18, pBHE2), and 85% of the odd straight chain FFAs was C15. However, the highest percentage of odd straight chain FFAs was achieved by the strain HWK201 (pXZ18, pBHE3) of 83.2% at 48 h. This strategy was also applied successfully in strains carrying different TE, such as the medium length acyl-ACP thioesterase gene from Umbellularia californica. C11 and C13 became the major odd straight-chain FFAs. © 2014 Wiley Periodicals, Inc.

  2. Long-term monitoring reveals carbon-nitrogen metabolism key to microcystin production in eutrophic lakes

    Lucas J Beversdorf

    2015-05-01

    Full Text Available The environmental drivers contributing to cyanobacterial dominance in aquatic systems have been extensively studied. However, understanding of toxic versus non-toxic cyanobacterial population dynamics and the mechanisms regulating cyanotoxin production remain elusive, both physiologically and ecologically. One reason is the disconnect between laboratory and field-based studies. Here, we combined three years of temporal data, including microcystin (MC concentrations, 16 years of long-term ecological research, and 10 years of molecular data to investigate the potential factors leading to the selection of toxic Microcystis and MC production. Our analysis revealed that nitrogen (N speciation and inorganic carbon (C availability might be important drivers of Microcystis population dynamics and that an imbalance in cellular C: N ratios may trigger MC production. More specifically, precipitous declines in ammonium concentrations lead to a transitional period of N stress, even in the presence of high nitrate concentrations, that we call the toxic phase. Following the toxic phase, temperature and cyanobacterial abundance remained elevated but MC concentrations drastically declined. Increases in ammonium due to lake turnover may have led to down regulation of MC synthesis or a shift in the community from toxic to non-toxic species. While total phosphorus (P to total N ratios were relatively low over the time-series, MC concentrations were highest when total N to total P ratios were also highest. Similarly, high C: N ratios were also strongly correlated to the toxic phase. We propose a metabolic model that corroborates molecular studies and reflects our ecological observations that C and N metabolism may regulate MC production physiologically and ecologically. In particular, we hypothesize that an imbalance between 2-oxoglutarate and ammonium in the cell regulates MC synthesis in the environment.

  3. Electrophoretic and zymographic techniques for production monitoring of two lipase forms from Candida antarctica DSM 70725

    Dimitrijević Aleksandra S.

    2012-01-01

    Full Text Available Yeast Candida antarctica produces two lipase forms, which are widely used as catalysts in variety of organic reactions, many of which are applied on a large scale. In this work, production of two forms of lipase from C. antarctica DSM 70725 (CAL A and CAL B was monitored during seven days of cultivation in the optimal medium using different electrophoretic and zymographic techniques. According to electrophoresis after silver staining, C. antarctica lipase A (molecular mass 45 kDa was produced starting from the second day of cultivation. C. antarctica lipase B (CAL B was also produced starting from the second day, but protein was present in the fermentation broth predominantly as dimer (molecular weight 66 kDa, while presence of monomeric form of CAL B (molecular weight of 33 kDa was observed starting from the fourth day of cultivation. Both types of zymograms (based on hydrolysis and synthesis reactions were used for detection of lipase activity in the fermentation broth. C. antarctica lipase A showed activity only in hydrolytic zymogram, when α-naphtyl butyrate was used as substrate. In the same zymogram, with α-naphtyl acetate as substrate no CAL A activity was detected. Similarly, CAL A showed no activity in synthesis based zymograms towards oleic acid and octanol as substrates, indicating that CAL A is not active towards very short or long-chain substrates. As opposite of CAL A, both monomeric and dimeric form of CAL B were detected in the all zymograms, suggesting that CAL B is active towards wide range of substrates, regardless to the chain length. Thus, zymogram based on hydrolysis of α-naphtyl butyrate represents a simple method for monitoring the production of two forms of lipase from C. antarctica, that greatly differ in their characteristics.

  4. Systems metabolic engineering of Corynebacterium glutamicum for production of the chemical chaperone ectoine.

    Becker, Judith; Schäfer, Rudolf; Kohlstedt, Michael; Harder, Björn J; Borchert, Nicole S; Stöveken, Nadine; Bremer, Erhard; Wittmann, Christoph

    2013-11-15

    The stabilizing and function-preserving effects of ectoines have attracted considerable biotechnological interest up to industrial scale processes for their production. These rely on the release of ectoines from high-salinity-cultivated microbial producer cells upon an osmotic down-shock in rather complex processor configurations. There is growing interest in uncoupling the production of ectoines from the typical conditions required for their synthesis, and instead design strains that naturally release ectoines into the medium without the need for osmotic changes, since the use of high-salinity media in the fermentation process imposes notable constraints on the costs, design, and durability of fermenter systems. Here, we used a Corynebacterium glutamicum strain as a cellular chassis to establish a microbial cell factory for the biotechnological production of ectoines. The implementation of a mutant aspartokinase enzyme ensured efficient supply of L-aspartate-beta-semialdehyde, the precursor for ectoine biosynthesis. We further engineered the genome of the basic C. glutamicum strain by integrating a codon-optimized synthetic ectABCD gene cluster under expressional control of the strong and constitutive C. glutamicum tuf promoter. The resulting recombinant strain produced ectoine and excreted it into the medium; however, lysine was still found as a by-product. Subsequent inactivation of the L-lysine exporter prevented the undesired excretion of lysine while ectoine was still exported. Using the streamlined cell factory, a fed-batch process was established that allowed the production of ectoine with an overall productivity of 6.7 g L(-1) day(-1) under growth conditions that did not rely on the use of high-salinity media. The present study describes the construction of a stable microbial cell factory for recombinant production of ectoine. We successfully applied metabolic engineering strategies to optimize its synthetic production in the industrial workhorse C

  5. Stimulation of acetoin production in metabolically engineered Lactococcus lactis by increasing ATP demand

    Liu, Jianming; Kandasamy, Vijayalakshmi; Würtz, Anders

    2016-01-01

    Having a sufficient supply of energy, usually in the form of ATP, is essential for all living organisms. In this study, however, we demonstrate that it can be beneficial to reduce ATP availability when the objective is microbial production. By introducing the ATP hydrolyzing F1-ATPase into a Lact...

  6. Effectiveness of Focus-on-Forms Instruction: Different Outcomes on Constrained- and Free-Production Tasks?

    Giselle Corbeil

    2005-06-01

    Full Text Available Abstract The purpose of this study is to investigate how focus-on-forms instruction in second language teaching affects attention to forms in two different types of task: constrained and free production. These two different types of task were administered to 87 university students enrolled in a first-year French as a second language course before and after instruction on qualifying adjective agreement. Comparisons were made between learning-gain differences from pre- to post-tests for both tasks. Significant differences between pre- and post-test scores were found for both tasks. Although the difference between the two tasks was not significant, the learning-gain difference was somewhat higher for the constrained-production task than for the free-production task. When the total number of adjectives used in the compositions was broken down into colour adjectives and non-colour adjectives, no significant learning gains were observed between the two categories, although the learning gains for the non-colour adjectives was twice as high as for the colour adjectives. Possible explanations for these results are discussed.

  7. Increased isobutanol production in Saccharomyces cerevisiae by overexpression of genes in valine metabolism

    Karhumaa Kaisa

    2011-07-01

    Full Text Available Abstract Background Isobutanol can be a better biofuel than ethanol due to its higher energy density and lower hygroscopicity. Furthermore, the branched-chain structure of isobutanol gives a higher octane number than the isomeric n-butanol. Saccharomyces cerevisiae was chosen as the production host because of its relative tolerance to alcohols, robustness in industrial fermentations, and the possibility for future combination of isobutanol production with fermentation of lignocellulosic materials. Results The yield of isobutanol was improved from 0.16 to 0.97 mg per g glucose by simultaneous overexpression of biosynthetic genes ILV2, ILV3, and ILV5 in valine metabolism in anaerobic fermentation of glucose in mineral medium in S. cerevisiae. Isobutanol yield was further improved by twofold by the additional overexpression of BAT2, encoding the cytoplasmic branched-chain amino-acid aminotransferase. Overexpression of ILV6, encoding the regulatory subunit of Ilv2, in the ILV2 ILV3 ILV5 overexpression strain decreased isobutanol production yield by threefold. In aerobic cultivations in shake flasks in mineral medium, the isobutanol yield of the ILV2 ILV3 ILV5 overexpression strain and the reference strain were 3.86 and 0.28 mg per g glucose, respectively. They increased to 4.12 and 2.4 mg per g glucose in yeast extract/peptone/dextrose (YPD complex medium under aerobic conditions, respectively. Conclusions Overexpression of genes ILV2, ILV3, ILV5, and BAT2 in valine metabolism led to an increase in isobutanol production in S. cerevisiae. Additional overexpression of ILV6 in the ILV2 ILV3 ILV5 overexpression strain had a negative effect, presumably by increasing the sensitivity of Ilv2 to valine inhibition, thus weakening the positive impact of overexpression of ILV2, ILV3, and ILV5 on isobutanol production. Aerobic cultivations of the ILV2 ILV3 ILV5 overexpression strain and the reference strain showed that supplying amino acids in cultivation media

  8. Glass Ceramic Waste Forms for Combined CS+LN+TM Fission Products Waste Streams

    Crum, Jarrod V.; Turo, Laura A.; Riley, Brian J.; Tang, Ming; Kossoy, Anna; Sickafus, Kurt E.

    2010-01-01

    In this study, glass ceramics were explored as an alternative waste form for glass, the current baseline, to be used for immobilizing alkaline/alkaline earth + lanthanide (CS+LN) or CS+LN+transition metal (TM) fission-product waste streams generated by a uranium extraction (UREX+) aqueous separations type process. Results from past work on a glass waste form for the combined CS+LN waste streams showed that as waste loading increased, large fractions of crystalline phases precipitated upon slow cooling.(1) The crystalline phases had no noticeable impact on the waste form performance by the 7-day product consistency test (PCT). These results point towards the development of a glass ceramic waste form for treating CS+LN or CS+LN+TM combined waste streams. Three main benefits for exploring glass ceramics are: (1) Glass ceramics offer increased solubility of troublesome components in crystalline phases as compared to glass, leading to increased waste loading; (2) The crystalline network formed in the glass ceramic results in higher heat tolerance than glass; and (3) These glass ceramics are designed to be processed by the same melter technology as the current baseline glass waste form. It will only require adding controlled canister cooling for crystallization into a glass ceramic waste form. Highly annealed waste form (essentially crack free) with up to 50X lower surface area than a typical High-Level Waste (HLW) glass canister. Lower surface area translates directly into increased durability. This was the first full year of exploring glass ceramics for the Option 1 and 2 combined waste stream options. This work has shown that dramatic increases in waste loading are achievable by designing a glass ceramic waste form as an alternative to glass. Table S1 shows the upper limits for heat, waste loading (based on solubility), and the decay time needed before treatment can occur for glass and glass ceramic waste forms. The improvements are significant for both combined waste

  9. Glass Ceramic Waste Forms for Combined CS+LN+TM Fission Products Waste Streams

    Crum, Jarrod V.; Turo, Laura A.; Riley, Brian J.; Tang, Ming; Kossoy, Anna; Sickafus, Kurt E.

    2010-09-23

    In this study, glass ceramics were explored as an alternative waste form for glass, the current baseline, to be used for immobilizing alkaline/alkaline earth + lanthanide (CS+LN) or CS+LN+transition metal (TM) fission-product waste streams generated by a uranium extraction (UREX+) aqueous separations type process. Results from past work on a glass waste form for the combined CS+LN waste streams showed that as waste loading increased, large fractions of crystalline phases precipitated upon slow cooling.[1] The crystalline phases had no noticeable impact on the waste form performance by the 7-day product consistency test (PCT). These results point towards the development of a glass ceramic waste form for treating CS+LN or CS+LN+TM combined waste streams. Three main benefits for exploring glass ceramics are: (1) Glass ceramics offer increased solubility of troublesome components in crystalline phases as compared to glass, leading to increased waste loading; (2) The crystalline network formed in the glass ceramic results in higher heat tolerance than glass; and (3) These glass ceramics are designed to be processed by the same melter technology as the current baseline glass waste form. It will only require adding controlled canister cooling for crystallization into a glass ceramic waste form. Highly annealed waste form (essentially crack free) with up to 50X lower surface area than a typical High-Level Waste (HLW) glass canister. Lower surface area translates directly into increased durability. This was the first full year of exploring glass ceramics for the Option 1 and 2 combined waste stream options. This work has shown that dramatic increases in waste loading are achievable by designing a glass ceramic waste form as an alternative to glass. Table S1 shows the upper limits for heat, waste loading (based on solubility), and the decay time needed before treatment can occur for glass and glass ceramic waste forms. The improvements are significant for both combined waste

  10. Studies on radiocalcium metabolism in pigs which suffer from a hereditary form of rickets (pseudo-vitamin D deficiency rickets)

    Grabe, C. von; Harmeyer, J.; Schmitt, R. (Veterinary School Hannover (Germany, F.R.). Inst. of Physiology; Veterinary School, Hannover (Germany, F.R.). Dept. of Medical Physics)

    1983-05-01

    Intestinal calcium absorption and calcium kinetics of plasma have been studied in pigs which suffer from pseudovitamin D deficiency rickets using /sup 45/Ca. On two consecutive days 200-250 ..mu..Ci of /sup 45/CaCl/sub 2/ was administered either orally or intravenously. The experiments were carried out in order to find out: (1) if kinetic measurements can provide suitable information on possible changes in calcium hemostasis during the development of the rachitic symptoms; (2) whether kinetic parameters of calcium metabolism can be used as indices to characterize the metabolic disturbance of calcium hemostasis.

  11. Studies on radiocalcium metabolism in pigs which suffer from a hereditary form of rickets (pseudo-vitamin D deficiency rickets)

    Grabe, C. von; Harmeyer, J.; Schmitt, R.; Veterinary School, Hannover

    1983-01-01

    Intestinal calcium absorption and calcium kinetics of plasma have been studied in pigs which suffer from pseudovitamin D deficiency rickets using 45 Ca. 200-250 μCi of 45 CaCl 2 administered either orally or intravenously on two consecutive days. The experiments were carried out in order to find out: (1) if kinetic measurements can provide suitable information on possible changes in calcium hemostasis during the development of the rachitic symptoms; (2) whether kinetic parameters of calcium metabolism can be used as indices to characterize the metabolic disturbance of calcium hemostasis. (author)

  12. Multi-omic profiling of EPO-producing Chinese hamster ovary cell panel reveals metabolic adaptation to heterologous protein production

    Ley, Daniel; Kazemi Seresht, Ali; Engmark, Mikael

    2015-01-01

    Chinese hamster ovary (CHO) cells are the preferred production host for many therapeutic proteins. The production of heterologous proteins in CHO cells imposes a burden on the host cell metabolism and impact cellular physiology on a global scale. In this work, a multi-omics approach was applied...

  13. Analysis of biotechnological hydrogen production efficiency for microorganisms with different types of metabolism, promising directions for further research

    Людмила Сергіївна Зубченко

    2015-10-01

    Full Text Available The basic biotechnological methods of hydrogen production, namely biological water splitting, using algae or cyanobacteria, dark fermentation and bioelectrochemical systems are analyzed in the article. Biochemical features of hydrogen production by microorganisms with different types of metabolism are reviewed. Disadvantages of each method and the prospects for future research are determined

  14. Production of 3-hydroxypropionic acid from glucose and xylose by metabolically engineered Saccharomyces cerevisiae

    Kanchana R. Kildegaard

    2015-12-01

    Full Text Available Biomass, the most abundant carbon source on the planet, may in the future become the primary feedstock for production of fuels and chemicals, replacing fossil feedstocks. This will, however, require development of cell factories that can convert both C6 and C5 sugars present in lignocellulosic biomass into the products of interest. We engineered Saccharomyces cerevisiae for production of 3-hydroxypropionic acid (3HP, a potential building block for acrylates, from glucose and xylose. We introduced the 3HP biosynthetic pathways via malonyl-CoA or β-alanine intermediates into a xylose-consuming yeast. Using controlled fed-batch cultivation, we obtained 7.37±0.17 g 3HP L−1 in 120 hours with an overall yield of 29±1% Cmol 3HP Cmol−1 xylose. This study is the first demonstration of the potential of using S. cerevisiae for production of 3HP from the biomass sugar xylose. Keywords: Metabolic engineering, Biorefineries, 3-hydroxypropionic acid, Saccharomyces cerevisiae, Xylose utilization

  15. Genetic and metabolic engineering for microbial production of poly-γ-glutamic acid.

    Cao, Mingfeng; Feng, Jun; Sirisansaneeyakul, Sarote; Song, Cunjiang; Chisti, Yusuf

    2018-05-28

    Poly-γ-glutamic acid (γ-PGA) is a natural biopolymer of glutamic acid. The repeating units of γ-PGA may be derived exclusively from d-glutamic acid, or l-glutamic acid, or both. The monomer units are linked by amide bonds between the α-amino group and the γ-carboxylic acid group. γ-PGA is biodegradable, edible and water-soluble. It has numerous existing and emerging applications in processing of foods, medicines and cosmetics. This review focuses on microbial production of γ-PGA via genetically and metabolically engineered recombinant bacteria. Strategies for improving production of γ-PGA include modification of its biosynthesis pathway, enhancing the production of its precursor (glutamic acid), and preventing loss of the precursor to competing byproducts. These and other strategies are discussed. Heterologous synthesis of γ-PGA in industrial bacterial hosts that do not naturally produce γ-PGA is discussed. Emerging trends and the challenges affecting the production of γ-PGA are reviewed. Copyright © 2018. Published by Elsevier Inc.

  16. Metabolic engineering of Pseudomonas putida KT2440 for the production of para-hydroxy benzoic acid

    Shiqin Yu

    2016-11-01

    Full Text Available para-hydroxy benzoic acid (PHBA is the key component for preparing parabens, a common preservatives in food, drugs and personal care products, as well as high performance bioplastics such as liquid crystal polymers (LCP. Pseudomonas putida KT2440 was engineered to produce PHBA from glucose via the shikimate pathway intermediate chorismate. To obtain the PHBA production strain, chorismate lyase UbiC from Escherichia coli and a feedback resistant 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase encoded by gene aroGD146N were overexpressed individually and simultaneously. In addition, genes related to product degradation (pobA or competing for the precursor chorismate (pheA and trpE were deleted from the genome. To further improve PHBA production, the glucose metabolism repressor hexR was knocked out in order to increase erythrose-4- phosphate and NAPH supply. The best strain achieved a maximum titre of 1.73 g L-1 and a carbon yield of 18.1 % (C-mol C-mol-1 in a non-optimized fed-batch fermentation. This is to date the highest PHBA concentration produced by P. putida using a chorismate lyase.

  17. Production of L-lactic acid from metabolically engineered strain of Enterobacter aerogenes ATCC 29007.

    Thapa, Laxmi Prasad; Lee, Sang Jun; Park, Chulhwan; Kim, Seung Wook

    2017-07-01

    In this study, L-lactic acid production was investigated from metabolically engineered strain of E. aerogenes ATCC 29007. The engineered strain E. aerogenes SUMI01 (Δpta) was generated by the deletion of phosphate acetyltransferase (pta) gene from the chromosome of E. aerogenes ATCC 29007 and deletion was confirmed by colony PCR. Under the optimized fermentation conditions, at 37°C and pH 6 for 84h, the L-lactic acid produced by engineered strain E. aerogenes SUMI01 (Δpta) in flask fermentation using 100g/L mannitol as the carbon source was 40.05g/L as compared to that of the wild type counterpart 20.70g/L. At the end of the batch fermentation in bioreactor the production of L-lactic acid reached to 46.02g/L and yield was 0.41g/g by utilizing 112.32g/L mannitol. This is the first report regarding the production of L-lactic acid from Enterobacter species. We believe that this result may provide valuable guidelines for further engineering Enterobacter strain for the improvement of L-lactic acid production. Copyright © 2017 Elsevier Inc. All rights reserved.

  18. Metabolic and mineral conditions of retained placenta in highly productive dairy cows: pathogenesis, diagnostics and prevention – a review

    Ryszard Mordak

    2017-01-01

    Full Text Available The time around calving in highly productive dairy cows is a critical period in terms of their metabolism, which is connected with high demands of the foetus as well as with the onset of lactation. Retained placenta in cows may have multifactorial aetiology, but in herds which are free from infectious diseases, the most important reasons are; periparturient metabolic changes and disturbances to the internal balance and stress. During the periparturient period, the most important factor causing immune suppression and hypotony of uterus in cows is metabolic stress due to hormonal and nutritional factors, including metabolic fluctuations, negative energy balance, as well as shortage of proteins, minerals, vitamins and antioxidants. This metabolic stress as a result of an imbalance in the internal metabolic homeostasis activates the hypothalamic-pituitary-adrenocortical axis (HPA and increases serum corticosterid (cortisol concentration, especially on the day of calving. Cortisol is a powerful immune suppressive factor that causes depression of leukocyte proliferation and their functions. The periparturient metabolic stress may also stimulate the production of catecholamines, especially adrenalin. Elevated levels of adrenalin activate adrenoreceptors of the myometrium, which in turn cause hypotony or atony of the uterus at calving in cows. Elevated levels of cortisol and adrenalin may significantly inhibit the rejection and expulsion of foetal membranes in cows, resulting in an increased incidence of their retention. These important mechanisms for placental retention in highly productive dairy cows often have primary nutritional metabolic aetiology, but they also occur during secondary metabolic disturbances and metabolic stress during calving. This metabolic and immunological aetiology and pathogenesis of retained placenta usually occur in highly productive periparturient cows on dairy farms in the absence of bovine infectious diseases, which can

  19. Metabolic and mineral conditions of retained placenta in highly productive dairy cows: pathogenesis, diagnostics and prevention – a review

    Ryszard Mordak; Józef Nicpoń; Josef Illek

    2017-01-01

    The time around calving in highly productive dairy cows is a critical period in terms of their metabolism, which is connected with high demands of the foetus as well as with the onset of lactation. Retained placenta in cows may have multifactorial aetiology, but in herds which are free from infectious diseases, the most important reasons are; periparturient metabolic changes and disturbances to the internal balance and stress. During the periparturient period, the most important factor causin...

  20. Heavy quark production form jet conversions in a quark-gluon plasma

    Liu , W.; Fries, R.

    2008-05-22

    Recently, it has been demonstrated that the chemical composition of jets in heavy ion collisions is significantly altered compared to the jets in the vacuum. This signal can be used to probe the medium formed in nuclear collisions. In this study we investigate the possibility that fast light quarks and gluons can convert to heavy quarks when passing through a quark-gluon plasma. We study the rate of light to heavy jet conversions in a consistent Fokker-Planck framework and investigate their impact on the production of high-p{sub T} charm and bottom quarks at the Relativistic Heavy Ion Collider and the Large Hadron Collider.

  1. Products of aqueous vitamin B5 (pantothenic acid) formed by free radical reactions

    Schittl, H. [Section of Radiation Biology, Department of Nutritional Sciences, University of Vienna, Althanstr. 14, UZA II, A-1090 Vienna (Austria); Quint, R.M. [Section of Radiation Biology, Department of Nutritional Sciences, University of Vienna, Althanstr. 14, UZA II, A-1090 Vienna (Austria); Getoff, N. [Section of Radiation Biology, Department of Nutritional Sciences, University of Vienna, Althanstr. 14, UZA II, A-1090 Vienna (Austria)]. E-mail: nikola.getoff@univie.ac.at

    2007-10-15

    The radiolysis of aqueous vitamin B5 (pantothenic acid) has been investigated under various experimental conditions. The highest vitamin degradation (G=3.22) was observed in solutions saturated with N{sub 2}O, where 90% OH radicals are operating. As final products, the following were established: aldehydes, carboxylic acids and ammonia. Their yield strongly depends on the presence/absence of air as well as on N{sub 2}O (used to convert e{sub aq} {sup -} into OH) and was determined as a function of absorbed radiation dose. HPLC-analysis showed that in all media, a main product is formed, having the highest yield in aerated solutions. Based on the chemical analysis, it appears that the OH radicals are most involved in the degradation process. A precise sequence of the reaction steps could not be given presently, because of the implication of many simultaneous reactions.

  2. Products of aqueous vitamin B5 (pantothenic acid) formed by free radical reactions

    Schittl, H.; Quint, R.M.; Getoff, N.

    2007-01-01

    The radiolysis of aqueous vitamin B5 (pantothenic acid) has been investigated under various experimental conditions. The highest vitamin degradation (G=3.22) was observed in solutions saturated with N 2 O, where 90% OH radicals are operating. As final products, the following were established: aldehydes, carboxylic acids and ammonia. Their yield strongly depends on the presence/absence of air as well as on N 2 O (used to convert e aq - into OH) and was determined as a function of absorbed radiation dose. HPLC-analysis showed that in all media, a main product is formed, having the highest yield in aerated solutions. Based on the chemical analysis, it appears that the OH radicals are most involved in the degradation process. A precise sequence of the reaction steps could not be given presently, because of the implication of many simultaneous reactions

  3. Numerical simulation of hot-pressed veneer products: Forming - Spring back – Distortion

    Ormarsson, Sigurdur; Sandberg, Dick

    2007-01-01

    Customers demand very high quality of veneered furniture products with regard to surface appearance, shape stability and stiffness. To meet these requirements, it is important to improve the manufacturing process by a better understanding of the thermo-hygro-mechanical behaviour of the individual...... veneers. During the manufacture of strongly curved products, the veneers are exposed to large membrane and bending deformations and to high pressure in the radial fibre direction. When hot-press forming is used, the veneers are also exposed to a high surface temperature during the pressing time (curing...... time). These severe conditions can result in plastic deformation perpendicular to the veneer surface as well as mechano-sorptive strains in the curved regions, since the heating can have a significant influence on the moisture distribution. How strong an influence these factors have on the distortion...

  4. Simulative Winding of Roll Formed Profile in Carcass Production for Flexible Pipes

    Nielsen, Peter Søe; Ormstrup, Casper Alexander; Hartz, Benjamin Arnold Krekeler

    2015-01-01

    numerous welds between coils of stainless steel, often duplex grades. The welds are a source of failure, since fracture from time to time occurs here in the winding stage. A simulative test in form of three-point-bending is developed, which shows promising results together with simplified air- and v......-bent profiles allowing offline testing of welds for optimisation purposes. Comparative studies are shown possible but discrepancies in boundary conditions cause the maximum strains in the simulative test to differ from those in production. A study of weld failure is done applying the simulative test and tensile...... tests using GOM ARAMIS 4M system for strain measurements. The results show strain localization at the weld from onset of yielding caused by the soft, heat affected zone next to the weld seam resulting in a local thinning of the strip similar to what is observed in production....

  5. Work efficiency improvement of >90% after implementation of an annual inpatient blood products administration consent form.

    Lindsay, Holly; Bhar, Saleh; Bonifant, Challice; Sartain, Sarah; Whittle, Sarah B; Lee-Kim, Youngna; Shah, Mona D

    2018-01-01

    Paediatric haematology, oncology and bone marrow transplant (BMT) patients frequently require transfusion of blood products. Our institution required a new transfusion consent be obtained every admission. The objectives of this project were to: revise inpatient blood products consent form to be valid for 1 year, decrease provider time spent consenting from 15 to improve provider frustration with the consent process. Over 6 months, we determined the average number of hospitalisations requiring transfusions in a random sampling of haematology/oncology/BMT inpatients. We surveyed nurses and providers regarding frustration levels and contact required regarding consents. Four and 12 months after implementation of the annual consent, providers and nurses were resurveyed, and new inpatient cohorts were assessed. Comparison of preintervention and postintervention time data allowed calculation of provider time reduction, a surrogate measure of improved work efficiency. Prior to the annual consent, >33 hours were spent over 6 months obtaining consent on 40 patients, with >19 hours spent obtaining consent when no transfusions were administered during admission. Twelve months after annual consent implementation, 97.5% (39/40) of analysed patients had a completed annual blood products transfusion consent and provider work efficiency had improved by 94.6% (>30 hours). Although several surveyed variables improved following annual consent implementation, provider frustration with consent process remained 6 out of a max score of 10, the same level as prior to the intervention. Development of an annual inpatient blood products consent form decreased provider time from 15 to 90%.

  6. 1,3-Propanediol dehydrogenases in Lactobacillus reuteri: impact on central metabolism and 3-hydroxypropionaldehyde production.

    Stevens, Marc J A; Vollenweider, Sabine; Meile, Leo; Lacroix, Christophe

    2011-08-03

    Lactobacillus reuteri metabolizes glycerol to 3-hydroxypropionaldehyde (3-HPA) and further to 1,3-propanediol (1,3-PDO), the latter step catalysed by a propanediol dehydrogenase (PDH). The last step in this pathway regenerates NAD+ and enables therefore the energetically more favourable production of acetate over ethanol during growth on glucose. A search throughout the genome of L. reuteri DSM 20016 revealed two putative PDHs encoded by ORFs lr_0030 and lr_1734. ORF lr_1734 is situated in the pdu operon encoding the glycerol conversion machinery and therefore likely involved in 1,3-PDO formation. ORF lr_0030 has not been associated with PDH-activity so far. To elucidate the role of these two PDHs, gene deletion mutant strains were constructed. Growth behaviour on glucose was comparable between the wild type and both mutant strains. However, on glucose + glycerol, the exponential growth rate of Δlr_0030 was lower compared to the wild type and the lr_1734 mutant. Furthermore, glycerol addition resulted in decreased ethanol production in the wild type and Δlr_1734, but not in Δlr_0030. PDH activity measurements using 3-HPA as a substrate revealed lower activity of Δlr_0030 extracts from exponential growing cells compared to wild type and Δlr_1734 extracts.During biotechnological 3-HPA production using non-growing cells, the ratio 3-HPA to 1,3-PDO was approximately 7 in the wild type and Δlr_0030, whereas this ratio was 12.5 in the mutant Δlr_1734. The enzyme encoded by lr_0030 plays a pivotal role in 3-HPA conversion in exponential growing L. reuteri cells. The enzyme encoded by lr_1734 is active during 3-HPA production by non-growing cells and this enzyme is a useful target to enhance 3-HPA production and minimize formation of the by-product 1,3-PDO.

  7. 1,3-Propanediol dehydrogenases in Lactobacillus reuteri: impact on central metabolism and 3-hydroxypropionaldehyde production

    Meile Leo

    2011-08-01

    Full Text Available Abstract Background Lactobacillus reuteri metabolizes glycerol to 3-hydroxypropionaldehyde (3-HPA and further to 1,3-propanediol (1,3-PDO, the latter step catalysed by a propanediol dehydrogenase (PDH. The last step in this pathway regenerates NAD+ and enables therefore the energetically more favourable production of acetate over ethanol during growth on glucose. Results A search throughout the genome of L. reuteri DSM 20016 revealed two putative PDHs encoded by ORFs lr_0030 and lr_1734. ORF lr_1734 is situated in the pdu operon encoding the glycerol conversion machinery and therefore likely involved in 1,3-PDO formation. ORF lr_0030 has not been associated with PDH-activity so far. To elucidate the role of these two PDHs, gene deletion mutant strains were constructed. Growth behaviour on glucose was comparable between the wild type and both mutant strains. However, on glucose + glycerol, the exponential growth rate of Δlr_0030 was lower compared to the wild type and the lr_1734 mutant. Furthermore, glycerol addition resulted in decreased ethanol production in the wild type and Δlr_1734, but not in Δlr_0030. PDH activity measurements using 3-HPA as a substrate revealed lower activity of Δlr_0030 extracts from exponential growing cells compared to wild type and Δlr_1734 extracts. During biotechnological 3-HPA production using non-growing cells, the ratio 3-HPA to 1,3-PDO was approximately 7 in the wild type and Δlr_0030, whereas this ratio was 12.5 in the mutant Δlr_1734. Conclusion The enzyme encoded by lr_0030 plays a pivotal role in 3-HPA conversion in exponential growing L. reuteri cells. The enzyme encoded by lr_1734 is active during 3-HPA production by non-growing cells and this enzyme is a useful target to enhance 3-HPA production and minimize formation of the by-product 1,3-PDO.

  8. Metabolic evolution and a comparative omics analysis of Corynebacterium glutamicum for putrescine production.

    Li, Zhen; Shen, Yu-Ping; Jiang, Xuan-Long; Feng, Li-Shen; Liu, Jian-Zhong

    2018-02-01

    Putrescine is widely used in the industrial production of bioplastics, pharmaceuticals, agrochemicals, and surfactants. Because the highest titer of putrescine is much lower than that of its precursor L-ornithine reported in microorganisms to date, further work is needed to increase putrescine production in Corynebacterium glutamicum. We first compared 7 ornithine decarboxylase genes and found that the Enterobacter cloacae ornithine decarboxylase gene speC1 was most suitable for putrescine production in C. glutamicum. Increasing NADPH availability and blocking putrescine oxidation and acetylation were chosen as targets for metabolic engineering. The putrescine producer C. glutamicum PUT4 was first constructed by deleting puo, butA and snaA genes, and replacing the fabG gene with E. cloacae speC1. After adaptive evolution with C. glutamicum PUT4, the evolved strain C. glutamicum PUT-ALE, which produced an 96% higher amount of putrescine compared to the parent strain, was obtained. The whole genome resequencing indicates that the SNPs located in the odhA coding region may be associated with putrescine production. The comparative proteomic analysis reveals that the pentose phosphate and anaplerotic pathway, the glyoxylate cycle, and the ornithine biosynthetic pathway were upregulated in the evolved strain C. glutamicum PUT-ALE. The aspartate family, aromatic, and branched chain amino acid and fatty acid biosynthetic pathways were also observed to be downregulated in C. glutamicum PUT-ALE. Reducing OdhA activity by replacing the odhA native start codon GTG with TTG and overexpression of cgmA or pyc458 further improved putrescine production. Repressing the carB, ilvH, ilvB and aroE expression via CRISPRi also increased putrescine production by 5, 9, 16 and 19%, respectively.

  9. Towards prediction of metabolic products of polyketide synthases: an in silico analysis.

    Gitanjali Yadav

    2009-04-01

    Full Text Available Sequence data arising from an increasing number of partial and complete genome projects is revealing the presence of the polyketide synthase (PKS family of genes not only in microbes and fungi but also in plants and other eukaryotes. PKSs are huge multifunctional megasynthases that use a variety of biosynthetic paradigms to generate enormously diverse arrays of polyketide products that posses several pharmaceutically important properties. The remarkable conservation of these gene clusters across organisms offers abundant scope for obtaining novel insights into PKS biosynthetic code by computational analysis. We have carried out a comprehensive in silico analysis of modular and iterative gene clusters to test whether chemical structures of the secondary metabolites can be predicted from PKS protein sequences. Here, we report the success of our method and demonstrate the feasibility of deciphering the putative metabolic products of uncharacterized PKS clusters found in newly sequenced genomes. Profile Hidden Markov Model analysis has revealed distinct sequence features that can distinguish modular PKS proteins from their iterative counterparts. For iterative PKS proteins, structural models of iterative ketosynthase (KS domains have revealed novel correlations between the size of the polyketide products and volume of the active site pocket. Furthermore, we have identified key residues in the substrate binding pocket that control the number of chain extensions in iterative PKSs. For modular PKS proteins, we describe for the first time an automated method based on crucial intermolecular contacts that can distinguish the correct biosynthetic order of substrate channeling from a large number of non-cognate combinatorial possibilities. Taken together, our in silico analysis provides valuable clues for formulating rules for predicting polyketide products of iterative as well as modular PKS clusters. These results have promising potential for discovery of

  10. Combining metabolic and process engineering strategies to improve recombinant glycoprotein production and quality.

    Karengera, Eric; Durocher, Yves; De Crescenzo, Gregory; Henry, Olivier

    2017-11-01

    Increasing recombinant protein production while ensuring a high and consistent protein quality remains a challenge in mammalian cell culture process development. In this work, we combined a nutrient substitution approach with a metabolic engineering strategy that improves glucose utilization efficiency. This combination allowed us to tackle both lactate and ammonia accumulation and investigate on potential synergistic effects on protein production and quality. To this end, HEK293 cells overexpressing the pyruvate yeast carboxylase (PYC2) and their parental cells, both stably producing the therapeutic glycoprotein interferon α2b (IFNα2b), were cultured in media deprived of glutamine but containing chosen substitutes. Among the tested substitutes, pyruvate led to the best improvement in growth (integral of viable cell density) for both cell lines in batch cultures, whereas the culture of PYC2 cells without neither glutamine nor any substitute displayed surprisingly enhanced IFNα2b production. The drastic reduction in both lactate and ammonia in the cultures translated into extended high viability conditions and an increase in recombinant protein titer by up to 47% for the parental cells and the PYC2 cells. Product characterization performed by surface plasmon resonance biosensing using Sambucus nigra (SNA) lectin revealed that the increase in yield was however accompanied by a reduction in the degree of sialylation of the product. Supplementing cultures with glycosylation precursors and a cofactor were effective at counterbalancing the lack of glutamine and allowed improvement in IFNα2b quality as evaluated by lectin affinity. Our study provides a strategy to reconcile protein productivity and quality and highlights the advantages of PYC2-overexpressing cells in glutamine-free conditions.

  11. Metabolic production of a novel polymer feedstock, 3-carboxy muconate, from vanillin.

    Gosling, Aaron; Fowler, S Jane; O'Shea, Michael S; Straffon, Melissa; Dumsday, Geoff; Zachariou, Michael

    2011-04-01

    Vanillin can be produced on a commercial scale by depolymerising renewable lignin. One product of microbial metabolism of vanillin by common soil microbes, such as Acinetobacter baylyi, is a tricarboxylic acid with a butadiene backbone known as 3-carboxy muconate (3CM). Three enzymes, 4-hydroxy benzaldehyde dehydrogenase, vanillate monooxygenase and protocatechuate 3,4-dioxygenase, catalyse the biotransformation of vanillin to 3CM. These three enzymes were metabolically engineered into an Escherichia coli host, giving a biocatalyst that converted vanillin into 3CM. The biocatalyst was found to give 100% yield of 3CM from 1 mM of vanillin after 39 h. The rate-limiting reaction was identified as the conversion of vanillate to 3,4-dihydroxybenzoate catalysed by vanillate monooxygenase. Low expression of the reductase subunit of this enzyme was identified as contributing to the reduced rate of this reaction. Proof of principle of a novel application for 3CM was demonstrated when it was converted into a trimethyl ester derivative and copolymerised with styrene.

  12. Harnessing the respiration machinery for high-yield production of chemicals in metabolically engineered Lactococcus lactis

    Liu, Jianming; Wang, Zhihao; Kandasamy, Vijayalakshmi

    2017-01-01

    on metabolically engineered Lactococcus lactis strains to optimize the production of acetoin and (R,R)−2,3-butanediol (R-BDO). In the absence of an external electron acceptor, a surplus of two NADH per acetoin molecule is produced. We found that a fully activated respiration was able to efficiently regenerate NAD......+, and a high titer of 371 mM (32 g/L) of acetoin was obtained with a yield of 82% of the theoretical maximum. Subsequently, we extended the metabolic pathway from acetoin to R-BDO by introducing the butanediol dehydrogenase gene from Bacillus subtilis. Since one mole of NADH is consumed when acetoin...... is converted into R-BDO per mole, only the excess of NADH needs to be oxidized via respiration. Either by fine-tuning the respiration capacity or by using a dual-phase fermentation approach involving a switch from fully respiratory to non-respiratory conditions, we obtained 361 mM (32 g/L) R-BDO with a yield...

  13. Indigenous bacteria and bacterial metabolic products in the gastrointestinal tract of broiler chickens.

    Rehman, Habib Ur; Vahjen, Wilfried; Awad, Wageha A; Zentek, Jürgen

    2007-10-01

    The gastrointestinal tract is a dynamic ecosystem containing a complex microbial community. In this paper, the indigenous intestinal bacteria and the microbial fermentation profile particularly short chain fatty acids (SCFA), lactate, and ammonia concentrations are reviewed. The intestinal bacterial composition changes with age. The bacterial density of the small intestine increases with age and comprises of lactobacilli, streptococci, enterobacteria, fusobacteria and eubacteria. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium spp., Clostridium spp., Lactobacillus spp., Fusobacterium spp. and Bacteroides) are predominating caecal bacteria in young broilers. Data from culture-based studies showed that bifidobacteria could not be isolated from young birds, but were recovered from four-week-old broilers. Caecal lactobacilli accounted for 1.5-24% of the caecal bacteria. Gene sequencing of caecal DNA extracts showed that the majority of bacteria belonged to Clostridiaceae. Intestinal bacterial community is influenced by the dietary ingredients, nutrient levels and physical structure of feed. SCFA and other metabolic products are affected by diet formulation and age. Additional studies are required to know the bacterial metabolic activities together with the community analysis of the intestinal bacteria. Feed composition and processing have great potential to influence the activities of intestinal bacteria towards a desired direction in order to support animal health, well-being and microbial safety of broiler meat.

  14. Lipid accumulation product is related to metabolic syndrome in women with polycystic ovary syndrome.

    Xiang, S; Hua, F; Chen, L; Tang, Y; Jiang, X; Liu, Z

    2013-02-01

    Metabolic disturbances are common features of polycystic ovary syndrome (PCOS), which possibly enhance the risk of diabetes and cardiovascular disease. Lipid accumulation product (LAP) is an emerging cardiovascular risk factor. The aim of this study was to explore the ability of LAP to identify metabolic syndrome (MS) in PCOS women. In a cross-sectional study, anthropometric, biochemical and clinical parameters were measured in 105 PCOS women. Receiver operating characteristic (ROC) analysis was used to find out the cut-off points of LAP to predict MS. MS was categorized according to International Diabetes Federation (IDF) criteria. The prevalence of MS was 43.8% in this study. PCOS women with MS had significantly higher LAP levels compared to those without MS. LAP was highly correlated with components of MS. ROC analysis showed that LAP was a significant discriminator for MS in PCOS women, and the optimal cutoff point of LAP to predict MS was 54.2 (93.3% sensitivity, 96.7% specificity). LAP seems to be associated with MS and has a strong and reliable diagnostic accuracy for MS in PCOS women. © J. A. Barth Verlag in Georg Thieme Verlag KG Stuttgart · New York.

  15. Quantitative Metabolomics and Instationary 13C-Metabolic Flux Analysis Reveals Impact of Recombinant Protein Production on Trehalose and Energy Metabolism in Pichia pastoris

    Joel Jordà

    2014-05-01

    Full Text Available Pichia pastoris has been recognized as an effective host for recombinant protein production. In this work, we combine metabolomics and instationary 13C metabolic flux analysis (INST 13C-MFA using GC-MS and LC-MS/MS to evaluate the potential impact of the production of a Rhizopus oryzae lipase (Rol on P. pastoris central carbon metabolism. Higher oxygen uptake and CO2 production rates and slightly reduced biomass yield suggest an increased energy demand for the producing strain. This observation is further confirmed by 13C-based metabolic flux analysis. In particular, the flux through the methanol oxidation pathway and the TCA cycle was increased in the Rol-producing strain compared to the reference strain. Next to changes in the flux distribution, significant variations in intracellular metabolite concentrations were observed. Most notably, the pools of trehalose, which is related to cellular stress response, and xylose, which is linked to methanol assimilation, were significantly increased in the recombinant strain.

  16. Some recent developments in sheet metal forming for production of lightweight automotive parts

    Tisza, M.; Lukács, Zs; Kovács, P.; Budai, D.

    2017-09-01

    Low cost manufacturing in the automotive industry is one of the main targets due to the ever increasing global competition among car manufacturers all over the World. Sheet metal forming is one of the most important key technologies in the automotive industry; therefore the elaboration of new, innovative low cost manufacturing processes is one of the main objectives in sheet metal forming as well. In 2015 with the initiative of the Imperial College London a research consortium was established under the umbrella Low Cost Materials Processing Technologies for Mass Production of Lightweight Vehicles. The primary aim of this project is to provide affordable low cost weight reduction in mass production of vehicles considering the entire life-cycle. In this project, 19 European Institutions (Universities and Research Institutions) from 9 European countries are participating with the above targets. The University of Miskolc is one of the members of this research Consortium. In this paper, some preliminary results with the contributions of the University of Miskolc will be introduced.

  17. Shape optimization of metal forming and forging products using the stress equivalent static loads calculated from a virtual model

    Jang, Hwan Hak; Jeong, Seong Beom; Park, Gyung Jin

    2012-01-01

    A shape optimization is proposed to obtain the desired final shape of forming and forging products in the manufacturing process. The final shape of a forming product depends on the shape parameters of the initial blank shape. The final shape of a forging product depends on the shape parameters of the billet shape. Shape optimization can be used to determine the shape of the blank and billet to obtain the appropriate final forming and forging products. The equivalent static loads method for non linear static response structural optimization (ESLSO) is used to perform metal forming and forging optimization since nonlinear dynamic analysis is required. Stress equivalent static loads (stress ESLs) are newly defined using a virtual model by redefining the value of the material properties. The examples in this paper show that optimization using the stress ESLs is quite useful and the final shapes of a forming and forging products are identical to the desired shapes

  18. METHODOLOGICAL INSTRUMENTS FOR FORMING THE MARKETING STRATEGY OF AGRICULTURAL PRODUCTION ECOLOGIZATION

    Mariya Bagorka

    2017-09-01

    Full Text Available The purpose of the article is the investigation of the nature, objectives, and strategic categories of ecologization marketing strategy, development of algorithm of this strategy and determination of environmental, economic, and ecological instruments of implementation of this strategy in the agricultural sector. Methodology. The basis of the formation of a marketing strategy for the ecologization of agrarian production is the systematic approach, which is based on the existence of implementation mechanisms that ensure system consistency, its purposefulness; interdependence; and complexity of its elements, which determines the integrity of the system; all tasks that execute individual elements of the system are interconnected; system elements and their associated actions have a certain subordination that builds a hierarchy; the system changes under the influence of specific factors, which determines its dynamism; the ability of the system to adapt to the variability of the external environment, while not losing its own individuality. Methods for strategic analysis of macro- and micro-environment factors were used to form the marketing strategy of ecologization. This process involves an analytical evaluation of the parameters of the external and internal environment with the help of general scientific and applied methods of strategic analysis. Results. The article explored the ecologization strategy, presented its objectives in the field of agriculture, the basic categories of strategic directions and implementation of ecologization marketing strategies. An algorithm of environmental marketing strategies in agricultural production is developed, environmental, economic and environmental and economic tools to implement them are determined. Practical implications. It is determined that marketing strategy of ecologization of agrarian production is a strategy of innovative development that can solve problems of economic growth, an increase of

  19. [Determination of sennosides and degraded products in the process of sennoside metabolism by HPLC].

    Sun, Yan; Li, Xuetuo; Yu, Xingju

    2004-01-01

    A method for the separation and determination of sennosides A and B and the main composition (sennidins A and B) in degraded products of sennosides by linear gradient high performance liquid chromatography has been developed. Separation conditions were as follows: column, a Spherisorb C18 column (250 mm x 4.6 mm i.d., 10 microm); column temperature, 40 degrees C; detection wavelength, 360 nm; mobile phase A, 1.25% acetic acid aqueous solution; mobile phase B, methanol; linear gradient, 100% A --> (20 min) 100% B. The method is effective, quick, accurate and reproducible. The satisfactory results show that this new method has certain practical values as an approach of real-time analysis in the process of sennoside metabolism.

  20. Enzyme and metabolic engineering for the production of novel biopolymers: crossover of biological and chemical processes.

    Matsumoto, Ken'ichiro; Taguchi, Seiichi

    2013-12-01

    The development of synthetic biology has transformed microbes into useful factories for producing valuable polymers and/or their precursors from renewable biomass. Recent progress at the interface of chemistry and biology has enabled the production of a variety of new biopolymers with properties that substantially differ from their petroleum-derived counterparts. This review touches on recent trials and achievements in the field of biopolymer synthesis, including chemo-enzymatically synthesized aliphatic polyesters, wholly biosynthesized lactate-based polyesters, polyhydroxyalkanoates and other unusual bacterially synthesized polyesters. The expanding diversities in structure and the material properties of biopolymers are key for exploring practical applications. The enzyme and metabolic engineering approaches toward this goal are discussed by shedding light on the successful case studies. Copyright © 2013 Elsevier Ltd. All rights reserved.

  1. [Clinical aspect of recent progress in phosphate metabolism. Distribution of phosphorus and its physiological roles in the body: the form, distribution, and physiological function].

    Yano, Shozo; Sugimoto, Toshitsugu

    2009-06-01

    Phosphorus plays pivotal roles in the survival such as the cellular structure, genomic information, energy metabolism, and cell signaling. Total amount of phosphorus is 500-700 g in human, most of which is stored in the bone in an insoluble form of calcium salt. About 15% of phosphorus is located in the cell membrane and the intracellular fluid in the soft tissues in a form of organic phosphate. Only 0.1% is present in the extracellular fluid. This phosphate pool plays a role in the dynamic equilibrium through the gut, kidney, bone and other tissues. Most of inorganic phosphates in the extracellular fluid are present in a form of ions such as H2PO4- and HPO(4)2-, and the concentration of phosphatic acids is about 1.2 mM. The form, distribution, and physiological function of phosphorus in the body are summarized in this review.

  2. Multifaceted metabolomics approaches for characterization of lignocellulosic biomass degradation products formed during ammonia fiber expansion pretreatment

    Vismeh, Ramin

    Lignocellulosic biomass represents a rather unused resource for production of biofuels, and it offers an alternative to food sources including corn starch. However, structural and compositional impediments limit the digestibility of sugar polymers in biomass cell walls. Thermochemical pretreatments improve accessibility of cellulose and hemicellulose to hydrolytic enzymes. However, most pretreatment methods generate compounds that either inhibit enzymatic hydrolysis or exhibit toxicity to fermentive microorganisms. Characterization and quantification of these products are essential for understanding chemistry of the pretreatment and optimizing the process efficiency to achieve higher ethanol yields. Identification of oligosaccharides released during pretreatment is also critical for choosing hydrolases necessary for cost-effective hydrolysis of cellulose and hemicellulose to fermentable monomeric sugars. Two chapters in this dissertation describe new mass spectrometry-based strategies for characterization and quantification of products that are formed during ammonia fiber expansion (AFEX) pretreatment of corn stover. Comparison of Liquid Chromatography Mass Spectrometry (LC/MS) profiles of AFEX-treated corn stover (AFEXTCS) and untreated corn stover (UTCS) extract shows that ammonolysis of lignin carbohydrate ester linkages generates a suite of nitrogenous compounds that are present only in the AFEXTCS extract and represent a loss of ammonia during processing. Several of these products including acetamide, feruloyl, coumaroyl and diferuloyl amides were characterized and quantified in the AFEXTCS extracts. The total amount of characterized and uncharacterized phenolic amides measured 17.4 mg/g AFEXTCS. Maillard reaction products including pyrazines and imidazoles were also identified and measured in the AFEXTCS extract totaling almost 1 mg/g AFEXTCS. The total of quantified nitrogenous products that are formed during AFEX was 43.4 mg/g AFEXTCS which was equivalent

  3. Single Point Incremental Forming to increase material knowledge and production flexibility

    Habraken, A.M.

    2016-01-01

    Nowadays, manufactured pieces can be divided into two groups: mass production and production of low volume number of parts. Within the second group (prototyping or small batch production), an emerging solution relies on Incremental Sheet Forming or ISF. ISF refers to processes where the plastic deformation occurs by repeated contact with a relatively small tool. More specifically, many publications over the past decade investigate Single Point Incremental Forming (SPIF) where the final shape is determined only by the tool movement. This manufacturing process is characterized by the forming of sheets by means of a CNC controlled generic tool stylus, with the sheets clamped by means of a non-workpiece-specific clamping system and in absence of a partial or a full die. The advantage is no tooling requirements and often enhanced formability, however it poses a challenge in term of process control and accuracy assurance. Note that the most commonly used materials in incremental forming are aluminum and steel alloys however other alloys are also used especially for medical industry applications, such as cobalt and chromium alloys, stainless steel and titanium alloys. Some scientists have applied incremental forming on PVC plates and other on sandwich panels composed of propylene with mild steel and aluminum metallic foams with aluminum sheet metal. Micro incremental forming of thin foils has also been developed. Starting from the scattering of the results of Finite Element (FE) simulations, when one tries to predict the tool force (see SPIF benchmark of 2014 Numisheet conference), we will see how SPIF and even micro SPIF (process applied on thin metallic sheet with a few grains within the thickness) allow investigating the material behavior. This lecture will focus on the identification of constitutive laws, on the SPIF forming mechanisms and formability as well as the failure mechanism. Different hypotheses have been proposed to explain SPIF formability, they will be

  4. Single Point Incremental Forming to increase material knowledge and production flexibility

    Habraken, A. M.

    2016-08-01

    Nowadays, manufactured pieces can be divided into two groups: mass production and production of low volume number of parts. Within the second group (prototyping or small batch production), an emerging solution relies on Incremental Sheet Forming or ISF. ISF refers to processes where the plastic deformation occurs by repeated contact with a relatively small tool. More specifically, many publications over the past decade investigate Single Point Incremental Forming (SPIF) where the final shape is determined only by the tool movement. This manufacturing process is characterized by the forming of sheets by means of a CNC controlled generic tool stylus, with the sheets clamped by means of a non-workpiece-specific clamping system and in absence of a partial or a full die. The advantage is no tooling requirements and often enhanced formability, however it poses a challenge in term of process control and accuracy assurance. Note that the most commonly used materials in incremental forming are aluminum and steel alloys however other alloys are also used especially for medical industry applications, such as cobalt and chromium alloys, stainless steel and titanium alloys. Some scientists have applied incremental forming on PVC plates and other on sandwich panels composed of propylene with mild steel and aluminum metallic foams with aluminum sheet metal. Micro incremental forming of thin foils has also been developed. Starting from the scattering of the results of Finite Element (FE) simulations, when one tries to predict the tool force (see SPIF benchmark of 2014 Numisheet conference), we will see how SPIF and even micro SPIF (process applied on thin metallic sheet with a few grains within the thickness) allow investigating the material behavior. This lecture will focus on the identification of constitutive laws, on the SPIF forming mechanisms and formability as well as the failure mechanism. Different hypotheses have been proposed to explain SPIF formability, they will be

  5. Metabolic engineering of Escherichia coli: a sustainable industrial platform for bio-based chemical production.

    Chen, Xianzhong; Zhou, Li; Tian, Kangming; Kumar, Ashwani; Singh, Suren; Prior, Bernard A; Wang, Zhengxiang

    2013-12-01

    In order to decrease carbon emissions and negative environmental impacts of various pollutants, more bulk and/or fine chemicals are produced by bioprocesses, replacing the traditional energy and fossil based intensive route. The Gram-negative rod-shaped bacterium, Escherichia coli has been studied extensively on a fundamental and applied level and has become a predominant host microorganism for industrial applications. Furthermore, metabolic engineering of E. coli for the enhanced biochemical production has been significantly promoted by the integrated use of recent developments in systems biology, synthetic biology and evolutionary engineering. In this review, we focus on recent efforts devoted to the use of genetically engineered E. coli as a sustainable platform for the production of industrially important biochemicals such as biofuels, organic acids, amino acids, sugar alcohols and biopolymers. In addition, representative secondary metabolites produced by E. coli will be systematically discussed and the successful strategies for strain improvements will be highlighted. Moreover, this review presents guidelines for future developments in the bio-based chemical production using E. coli as an industrial platform. Copyright © 2013 Elsevier Inc. All rights reserved.

  6. Lipid metabolism and potentials of biofuel and high added-value oil production in red algae.

    Sato, Naoki; Moriyama, Takashi; Mori, Natsumi; Toyoshima, Masakazu

    2017-04-01

    Biomass production is currently explored in microalgae, macroalgae and land plants. Microalgal biofuel development has been performed mostly in green algae. In the Japanese tradition, macrophytic red algae such as Pyropia yezoensis and Gelidium crinale have been utilized as food and industrial materials. Researches on the utilization of unicellular red microalgae such as Cyanidioschyzon merolae and Porphyridium purpureum started only quite recently. Red algae have relatively large plastid genomes harboring more than 200 protein-coding genes that support the biosynthetic capacity of the plastid. Engineering the plastid genome is a unique potential of red microalgae. In addition, large-scale growth facilities of P. purpureum have been developed for industrial production of biofuels. C. merolae has been studied as a model alga for cell and molecular biological analyses with its completely determined genomes and transformation techniques. Its acidic and warm habitat makes it easy to grow this alga axenically in large scales. Its potential as a biofuel producer is recently documented under nitrogen-limited conditions. Metabolic pathways of the accumulation of starch and triacylglycerol and the enzymes involved therein are being elucidated. Engineering these regulatory mechanisms will open a possibility of exploiting the full capability of production of biofuel and high added-value oil. In the present review, we will describe the characteristics and potential of these algae as biotechnological seeds.

  7. Nitrogen metabolism and protozoa production rate in cattle fed on diet containing protected protein

    Singh, G.P.; Gupta, B.N.

    1992-01-01

    Nitrogen metabolism and protozoa production rate using 14 C-choline as marker were studied on 9 adult male crossbred (Tharparker x Brown Swiss) rumen fistulated animals divided into 3 groups (A, B and C). All the animals were fed concentrate mixture and wheatstraw. However, groundnut cake (GNC) in concentrate mixture was untreated in group A, 50 per cent formaldehyde treated in group B and 100 per cent formaldehyde treated in group C. Although, DM intake was similar in these groups but water intake was significantly (P<0.05) higher in control group. Total-N, ammonia-N and blood urea were significantly lower in group B and C as compared to group A. Apparent CP digestibility was not affected by addition of formaldehyde treated GNC at 50 and 100 per cent levels. However, N balances increased significantly (P<0.05) due to addition of protected protein in diet. Protozoal pool as well as production rate were significantly (P<0.01) decreased due to formaldehyde treatment of GNC protein. Thus addition of formaldehyde treated GNC in diets decreased ammonia and protozoa production but increased N retention in groups B and C. (author). 27 refs., 3 tabs., 2 figs

  8. Metabolic flux analysis of the hydrogen production potential in Synechocystis sp. PCC6803

    Navarro, E. [Departamento de Lenguajes y Ciencias de la Computacion, Campus de Teatrinos, Universidad de Malaga, 29071 Malaga (Spain); Montagud, A.; Fernandez de Cordoba, P.; Urchueguia, J.F. [Instituto Universitario de Matematica Pura y Aplicada, Universidad Politecnica de Valencia, Camino de Vera 14, 46022 Valencia (Spain)

    2009-11-15

    Hydrogen is a promising energy vector; however, finding methods to produce it from renewable sources is essential to allow its wide-scale use. In that line, biological hydrogen production, although it is considered as a possible alternative, requires substantial improvements to overcome its present low yields. In that direction, genetic manipulation probably will play a central role and from that point of view metabolic flux analysis (MFA) constitutes an important tool to guide a priori most suitable genetic modifications oriented to a hydrogen yield increase. In this work MFA has been applied to analyze hydrogen photoproduction of Synechocystis sp. PCC6803. Flux analysis was carried out based on literature data and several basic fluxes were estimated in different growing conditions of the system. From this analysis, an upper limit for hydrogen photoproduction has been determined indicating a wide margin for improvement. MFA was also used to find a feasible operating space for hydrogen production, which avoids oxygen inhibition, one of the most important limitations to make hydrogen production cost effective. In addition, a set of biotechnological strategies are proposed that would be consistent with the performed mathematical analysis. (author)

  9. L-Cysteine Production in Escherichia coli Based on Rational Metabolic Engineering and Modular Strategy.

    Liu, Han; Fang, Guochen; Wu, Hui; Li, Zhimin; Ye, Qin

    2018-05-01

    L-cysteine is an amino acid with important physiological functions and has a wide range of applications in medicine, food, animal feed, and cosmetics industry. In this study, the L-cysteine synthesis in Escherichia coliEscherichia coli is divided into four modules: the transport module, sulfur module, precursor module, and degradation module. The engineered strain LH03 (overexpression of the feedback-insensitive cysE and the exporter ydeD in JM109) accumulated 45.8 mg L -1 of L-cysteine in 48 hr with yield of 0.4% g/g glucose. Further modifications of strains and culture conditions which based on the rational metabolic engineering and modular strategy improved the L-cysteine biosynthesis significantly. The engineered strain LH06 (with additional overexpression of serA, serC, and serB and double mutant of tnaA and sdaA in LH03) produced 620.9 mg L -1 of L-cysteine with yield of 6.0% g/g glucose, which increased the production by 12 times and the yield by 14 times more than those of LH03 in the original condition. In fed-batch fermentation performed in a 5-L reactor, the concentration of L-cysteine achieved 5.1 g L -1 in 32 hr. This work demonstrates that the combination of rational metabolic engineering and module strategy is a promising approach for increasing the L-cysteine production in E. coli. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Production and chemical composition of grasses and legumes cultivated in pure form, mixed or in consortium

    Antonio Augusto Cortiana Tambara

    2017-07-01

    Full Text Available The objective of this study was to assess the edible biomass and chemical composition of forages grown on pure form, as a grass mix, and in grass-legume consortia. The following species were tested: white oats (Avena sativa, black oats (Avena strigosa, ryegrass (Lolium multiflorum, forage peanut (Arachis pintoi, white clover (Trifolium repens, and red clover (Trifolium pratense. The experiment consisted of sixteen treatments arranged in a completely randomized design. The parameters measured were total dry matter (PMST, neutral detergent fiber (NDF, acid detergent fiber (ADF, and crude protein (CP. No significant differences in PMST were found among the consortia (p > 0.05. Only the pure cultivated white clover (p > 0.05 was comparable to the consortia in terms of biomass production. The three legumes had the lowest average NDF values (p > 0.05, based on their contributions to the total NDF content of the consortia along the cuts. The ADF content increased for all treatments during the cuts. The results indicate that in pasture, legumes increase protein content, and forage consortia increase both the pasture production and the grazing period. Their chemical composition is adequate for boosting livestock production in pastures.

  11. Catabolism of biomass-derived sugars in fungi and metabolic engineering as a tool for organic acid production

    Koivistoinen, O.

    2013-11-01

    gene ladB was identified and the deletion of the gene resulted in growth arrest on galactitol indicating that the enzyme is an essential part of the oxido-reductive galactose pathway in fungi. The last step of this pathway converts D-sorbitol to D-fructose by sorbitol dehydrogenase encoded by sdhA gene. Sorbitol dehydrogenase was found to be a medium chain dehydrogenase and transcription analysis suggested that the enzyme is involved in D-galactose and D-sorbitol catabolism. The thesis also demonstrates how the understanding of cell metabolism can be used to engineer yeast to produce glycolic acid. Glycolic acid is a chemical, which can be used for example in the cosmetic industry and as a precursor for biopolymers. Currently, glycolic acid is produced by chemical synthesis in a process requiring toxic formaldehyde and fossil fuels. Thus, a biochemical production route would be preferable from a sustainability point of view. Yeasts do not produce glycolic acid under normal conditions but it is a desired production host for acid production because of its natural tolerance to low pH conditions. As a proof of concept, pure model substrates, e.g. D-xylose and ethanol, were used as starting materials for glycolic acid production but the knowledge can be further applied to an expanded substrate range such as biomass derived sugars. Already the introduction of a heterologous glyoxylate reductase gene resulted in glycolic acid production in the yeasts S. cerevisiae and Kluyveromyces lactis. Further modifications of the glyoxylate cycle increased the production of glycolic acid and it was successfully produced in bioreactor cultivation. The challenge of biotechnology is to produce high value products from cheap raw materials in an economically feasible way. This thesis gives more basic understanding to the topic in the form of new information regarding L-rhamnose and D-galactose metabolism in eukaryotic microbes as well as provides an example on how cell metabolism can be

  12. Release of Phosphorus Forms from Cover Crop Residues in Agroecological No-Till Onion Production

    Rodolfo Assis de Oliveira

    Full Text Available ABSTRACT Cover crops grown alone or in association can take up different amounts of phosphorus (P from the soil and accumulate it in different P-forms in plant tissue. Cover crop residues with a higher content of readily decomposed forms may release P more quickly for the next onion crop. The aim of this study was to evaluate the release of P forms from residues of single and mixed cover crops in agroecological no-till onion (Allium cepa L. production. The experiment was conducted in Ituporanga, Santa Catarina (SC, Brazil, in an Inceptisol, with the following treatments: weeds, black oat (Avena sativa L., rye (Secale cereale L., oilseed radish (Raphanus sativus L., oilseed radish + black oat, and oilseed radish + rye. Cover crops were sown in April 2013. In July 2013, plant shoots were cut close to the soil surface and part of the material was placed in litterbags. The bags were placed on the soil surface and residues were collected at 0, 15, and 45 days after deposition (DAD. Residues were dried and ground and P in the plant tissue was determined through chemical fractionation. The release of P contained in the tissue of cover crops depends not only on total P content in the tissue, but also on the accumulation of P forms and the quality of the residue in decomposition. The highest accumulation of P in cover crops occurred in the soluble inorganic P fraction, which is the fraction of fastest release in plants. Black oat had the highest initial release rate of soluble inorganic P, which became equal to the release rate of other cover crop residues at 45 DAD. Weeds released only half the amount of soluble inorganic P in the same period, despite accumulating a considerable amount of P in their biomass. The mixtures of oilseed radish + rye and oilseed radish + black oat showed higher release of P associated with RNA at 45 DAD in comparison to the single treatments.

  13. Integrating biocompatible chemistry and manipulating cofactor partitioning in metabolically engineeredLactococcus lactisfor fermentative production of (3S)-acetoin

    Liu, Jianming; Solem, Christian; Jensen, Peter Ruhdal

    2016-01-01

    Biocompatible chemistry (BC), i.e. non-enzymatic chemical reactions compatible with living organisms, is increasingly used in conjunction with metabolically engineered microorganisms for producing compounds that do not usually occur naturally. Here we report production of one such compound, (3S......)-acetoin, a valuable precursor for chiral synthesis, using a metabolically engineered Lactococcus lactis strain growing under respiratory conditions with ferric iron serving as a BC component. The strain used has all competing product pathways inactivated, and an appropriate cofactor balance is achieved by fine...

  14. [Proceeding: Production rate, metabolic clearance rate and mean plasma concentration of cortisol in hyperthyroidism (author's transl)].

    Linquette, M; Lefebvre, J; Racadot, A; Cappoen, J P

    1975-01-01

    The adrenocortical function was studied in 23 patients with hyperthyroidism and compared with a group of 15 normal subjects. Parameters of adrenal function were determined with 1,2(3)H-cortisol. The half-life of cortisol is significantly shortened in hyperthyroidism, as compared to normal subjects (49,5 +/- 6,6 min vs 68,3 +/- 10,5 min) and metabolic clearance rate is increased (418,5 +/- 89,5 L/24 h vs 237,5 +/- 48,5 L/24 h, for normal subjects). The production rate of cortisol, calculated from specific and cumulate activities of THE and THF is increased in hyperthyroidism expressed as mg/24 h or mg/m2/24 h (respectively : 26,7 +/- 7,8 mg/24 h vs 15,7 +/- 3 mg/24 h and 16,9 +/- 4,6 mg/m2/24 h vs 9,5 +/- 1,8 mg/m2/24 h). The mean plasma concentration, calculated as the radio (see article) is not statiscally different in hyperthyroid and normal subjects (6,8 +/- 2,1 microg/100 ml vs 7,3 +/- 1,9 microg/100 ml). 7 patients were reinvestigated after treatment of thyrotoxicosis when they were clinically and biologically in euthyroid state. All the values were normalized, without statistically significant difference from control (T 1/2 = 65,4 +/- 18 min, Metb Cl. Rate : 255 +/- 64,5 L/24 h, production rate : 15,6 +/- 1,8 mg/24 h and 9 +/- 1,4 mg/m2/24 h. mean plasma concentration : 6,8 +/- 2,8 microg/100 ml). Shortened cortisol half life, increased metabolic clearance rate and production rate, and normal mean plasma concentration have been reported in hyperthyroidism (Peterson, Copinschi, Gallagher). These changes, secondary to thyroid hormone excess, are the consequences of increased hepatic catabolism of cortisol. The activity of 11 OH steroid deshydrogenase is increased, as demonstrated by increased ratio (see article) in normal subjects (0,001 less than p less than 0,005). There is a high proportion of 17 kéto metabolites (E, DHE, THE) whose feed-back effect is weak as compared to 17 OH metabolites (F, DHF, THF). The hypothalamo-hypophyso-adrenal system is

  15. Dysregulated metabolism contributes to oncogenesis

    Hirschey, Matthew D.; DeBerardinis, Ralph J.; Diehl, Anna Mae E.; Drew, Janice E.; Frezza, Christian; Green, Michelle F.; Jones, Lee W.; Ko, Young H.; Le, Anne; Lea, Michael A.; Locasale, Jason W.; Longo, Valter D.; Lyssiotis, Costas A.; McDonnell, Eoin; Mehrmohamadi, Mahya; Michelotti, Gregory; Muralidhar, Vinayak; Murphy, Michael P.; Pedersen, Peter L.; Poore, Brad; Raffaghello, Lizzia; Rathmell, Jeffrey C.; Sivanand, Sharanya; Vander Heiden, Matthew G.; Wellen, Kathryn E.

    2015-01-01

    Cancer is a disease characterized by unrestrained cellular proliferation. In order to sustain growth, cancer cells undergo a complex metabolic rearrangement characterized by changes in metabolic pathways involved in energy production and biosynthetic processes. The relevance of the metabolic transformation of cancer cells has been recently included in the updated version of the review “Hallmarks of Cancer”, where the dysregulation of cellular metabolism was included as an emerging hallmark. While several lines of evidence suggest that metabolic rewiring is orchestrated by the concerted action of oncogenes and tumor suppressor genes, in some circumstances altered metabolism can play a primary role in oncogenesis. Recently, mutations of cytosolic and mitochondrial enzymes involved in key metabolic pathways have been associated with hereditary and sporadic forms of cancer. Together, these results suggest that aberrant metabolism, once seen just as an epiphenomenon of oncogenic reprogramming, plays a key role in oncogenesis with the power to control both genetic and epigenetic events in cells. In this review, we discuss the relationship between metabolism and cancer, as part of a larger effort to identify a broad-spectrum of therapeutic approaches. We focus on major alterations in nutrient metabolism and the emerging link between metabolism and epigenetics. Finally, we discuss potential strategies to manipulate metabolism in cancer and tradeoffs that should be considered. More research on the suite of metabolic alterations in cancer holds the potential to discover novel approaches to treat it. PMID:26454069

  16. Fatty Liver Index and Lipid Accumulation Product Can Predict Metabolic Syndrome in Subjects without Fatty Liver Disease

    Yuan-Lung Cheng

    2017-01-01

    Full Text Available Background. Fatty liver index (FLI and lipid accumulation product (LAP are indexes originally designed to assess the risk of fatty liver and cardiovascular disease, respectively. Both indexes have been proven to be reliable markers of subsequent metabolic syndrome; however, their ability to predict metabolic syndrome in subjects without fatty liver disease has not been clarified. Methods. We enrolled consecutive subjects who received health check-up services at Taipei Veterans General Hospital from 2002 to 2009. Fatty liver disease was diagnosed by abdominal ultrasonography. The ability of the FLI and LAP to predict metabolic syndrome was assessed by analyzing the area under the receiver operating characteristic (AUROC curve. Results. Male sex was strongly associated with metabolic syndrome, and the LAP and FLI were better than other variables to predict metabolic syndrome among the 29,797 subjects. Both indexes were also better than other variables to detect metabolic syndrome in subjects without fatty liver disease (AUROC: 0.871 and 0.879, resp., and the predictive power was greater among women. Conclusion. Metabolic syndrome increases the cardiovascular disease risk. The FLI and LAP could be used to recognize the syndrome in both subjects with and without fatty liver disease who require lifestyle modifications and counseling.

  17. Solar photocatalytic oxidation of recalcitrant natural metabolic by-products of amoxicillin biodegradation

    João Pereira; Ana Reis; Vera Homem; José Silva; Arminda Alves; Maria Teresa Borges; Rui Boaventura; Vítor Vilar; Olga Pastor Nunes

    2014-01-01

    The contamination of the aquatic environment by non-metabolized and metabolized antibiotic residues has brought the necessity of alternative treatment steps to current water decontamination technologies. This work assessed the feasibility of using a multi-stage treatment system for amoxicillin (AMX) spiked solutions combining: i) a biological treatment process using an enriched culture to metabolize AMX, with ii) a solar photocatalytic system to achieve the removal of the metabolized transfor...

  18. Forms and methods of stimulation of innovative activities in the restructuring of production program

    I. I. Emtcova

    2016-01-01

    Full Text Available In the Russian economy not every business entity, implements innovative business activities. The situation generated by the complexity of perception and practical transition to an innovative economic system. On the development of innovative activities affects the overall condition of the economy, condition of material production. The research demonstrates that resource potential of innovative activities in recent years had a tendency towards absolute quantitative reduction and quality deterioration. The decrease in the level and quality of resource provision of innovative activity due to the lack of necessary financial resources. Currently, innovation has become the primary means of increasing the profit of economic entities at the expense of better meet market demand, reduce production costs compared to competitors. Given the complexity of businesses, there is a need of the state stimulation of innovative activity, which is carried out the main directions, forms and methods. In the system of direct effects of the state on business innovation is the stimulation of development of Technopark structures. Creating the most favourable conditions for innovative enterprises, the provision of various services is their main goal. For the food processing industry currently, the largest share in the investments in the investment activities have their own sources of funding, including the use of depreciation. To Finance industry-wide, cross-sectoral and regional scientific and technical problems you can create extra-budgetary funds for financing R & d and innovation support. To encourage regional interests, one of which is that innovation is available to local authorities. In the financial provision of innovative activity is given credit. A Bank loan allows you to increase the efficiency of innovation activity. The article concludes that these measures to stimulate innovative-innovative activity can effectively influence the activity of the company: will

  19. Two phosphoenolpyruvate carboxykinases coexist in the Crassulacean Acid Metabolism plant Ananas comosus. Isolation and characterization of the smaller 65 kDa form.

    Martín, Mariana; Rius, Sebastián Pablo; Podestá, Florencio Esteban

    2011-06-01

    Two phosphoenolpyruvate carboxykinase (PEPCK, EC 4.1.1.49) isoforms of 74 and 65 kDa were found to coexist in vivo in pineapple leaves, a constitutive Crassulacean Acid Metabolism plant. The 65 kDa form was not the result of proteolytic cleavage of the larger form since extraction methods reported to prevent PEPCK proteolysis in other plant tissues failed to yield a single immunoreactive PEPCK polypeptide in leaf extracts. In this work, the smaller form of 65 kDa was purified to homogeneity and physically and kinetically characterized and showed parameters compatible with a fully active enzyme. The specific activity was nearly twice higher for decarboxylation of oxaloacetate when compared to carboxylation of phosphoenolpyruvate. Kinetic parameters fell within the range of those estimated for other plant PEPCKs. Its activity was affected by several metabolites, as shown by inhibition by 3-phosphoglycerate, citrate, malate, fructose-1,6-bisphosphate, l-asparagine and activation of the decarboxylating activity by succinate. A break in the Arrhenius plot at about 30°C indicates that PEPCK structure is responsive to changes in temperature. The results indicate that pineapple leaves contain two PEPCK forms. The biochemical characterization of the smaller isoform performed in this work suggests that it could participate in both carbon and nitrogen metabolism in vivo by acting as a decarboxylase. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  20. Metabolic engineering of lactic acid bacteria for the production of nutraceuticals

    Hugenholtz, J.; Sybesma, W.; Groot, M.N.; Wisselink, W.; Ladero, V.; Burgess, K.; Sinderen, van D.; Piard, J.C.; Eggink, G.; Smid, E.J.; Savoy, G.; Sesma, F.; Jansen, T.; Hols, P.; Kleerebezem, M.

    2002-01-01

    Lactic acid bacteria display a relatively simple and well-described metabolism where the sugar source is converted mainly to lactic acid. Here we will shortly describe metabolic engineering strategies on the level of sugar metabolism, that lead to either the efficient re-routing of the lactococcal

  1. Camelina sativa: An ideal platform for the metabolic engineering and field production of industrial lipids.

    Bansal, Sunil; Durrett, Timothy P

    2016-01-01

    Triacylglycerols (TAG) containing modified fatty acids with functionality beyond those found in commercially grown oil seed crops can be used as feedstocks for biofuels and bio-based materials. Over the years, advances have been made in transgenically engineering the production of various modified fatty acids in the model plant Arabidopsis thaliana. However, the inability to produce large quantities of transgenic seed has limited the functional testing of the modified oil. In contrast, the emerging oil seed crop Camelina sativa possesses important agronomic traits that recommend it as an ideal production platform for biofuels and industrial feedstocks. Camelina possesses low water and fertilizer requirements and is capable of yields comparable to other oil seed crops, particularly under stress conditions. Importantly, its relatively short growing season enables it to be grown as part of a double cropping system. In addition to these valuable agronomic features, Camelina is amenable to rapid metabolic engineering. The development of a simple and effective transformation method, combined with the availability of abundant transcriptomic and genomic data, has allowed the generation of transgenic Camelina lines capable of synthesizing high levels of unusual lipids. In some cases these levels have surpassed what was achieved in Arabidopsis. Further, the ability to use Camelina as a crop production system has allowed for the large scale growth of transgenic oil seed crops, enabling subsequent physical property testing. The application of new techniques such as genome editing will further increase the suitability of Camelina as an ideal platform for the production of biofuels and bio-materials. Copyright © 2015 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

  2. *NO and oxyradical metabolism in new cell lines of rat brain capillary endothelial cells forming the blood-brain barrier.

    Blasig, I E; Giese, H; Schroeter, M L; Sporbert, A; Utepbergenov, D I; Buchwalow, I B; Neubert, K; Schönfelder, G; Freyer, D; Schimke, I; Siems, W E; Paul, M; Haseloff, R F; Blasig, R

    2001-09-01

    To investigate the relevance of *NO and oxyradicals in the blood-brain barrier (BBB), differentiated and well-proliferating brain capillary endothelial cells (BCEC) are required. Therefore, rat BCEC (rBCEC) were transfected with immortalizing genes. The resulting lines exhibited endothelial characteristics (factor VIII, angiotensin-converting enzyme, high prostacyclin/thromboxane release rates) and BBB markers (gamma-glutamyl transpeptidase, alkaline phosphatase). The control line rBCEC2 (mock transfected) revealed fibroblastoid morphology, less factor VIII, reduced gamma-glutamyl transpeptidase, weak radical defence, low prostanoid metabolism, and limited proliferation. Lines transfected with immortalizing genes (especially rBCEC4, polyoma virus large T antigen) conserved primary properties: epitheloid morphology, subcultivation with high proliferation rate under pure culture conditions, and powerful defence against reactive oxygen species (Mn-, Cu/Zn-superoxide dismutase, catalase, glutathione peroxidase, glutathione) effectively controlling radical metabolism. Only 100 microM H2O2 overcame this defence and stimulated the formation of eicosanoids similarly as in primary cells. Some BBB markers were expressed to a lower degree; however, cocultivation with astrocytes intensified these markers (e.g., alkaline phosphatase) and paraendothelial tightness, indicating induction of BBB properties. Inducible NO synthase was induced by a cytokine plus lipopolysaccharide mixture in all lines and primary cells, resulting in *NO release. Comparing the cell lines obtained, rBCEC4 are stable immortalized and reveal the best conservation of properties from primary cells, including enzymes producing or decomposing reactive species. These cells can be subcultivated in large amounts and, hence, they are suitable to study the role of radical metabolism in the BBB and in the cerebral microvasculature. Copyright 2001 Academic Press.

  3. Vanillin production using metabolically engineered Escherichia coli under non-growing conditions.

    Barghini, Paolo; Di Gioia, Diana; Fava, Fabio; Ruzzi, Maurizio

    2007-04-16

    Vanillin is one of the most important aromatic flavour compounds used in the food and cosmetic industries. Natural vanillin is extracted from vanilla beans and is relatively expensive. Moreover, the consumer demand for natural vanillin highly exceeds the amount of vanillin extracted by plant sources. This has led to the investigation of other routes to obtain this flavour such as the biotechnological production from ferulic acid. Studies concerning the use of engineered recombinant Escherichia coli cells as biocatalysts for vanillin production are described in the literature, but yield optimization and biotransformation conditions have not been investigated in details. Effect of plasmid copy number in metabolic engineering of E. coli for the synthesis of vanillin has been evaluated by the use of genes encoding feruloyl-CoA synthetase and feruloyl hydratase/aldolase from Pseudomonas fluorescens BF13. The higher vanillin production yield was obtained using resting cells of E. coli strain JM109 harbouring a low-copy number vector and a promoter exhibiting a low activity to drive the expression of the catabolic genes. Optimization of the bioconversion of ferulic acid to vanillin was accomplished by a response surface methodology. The experimental conditions that allowed us to obtain high values for response functions were 3.3 mM ferulic acid and 4.5 g/L of biomass, with a yield of 70.6% and specific productivity of 5.9 micromoles/g x min after 3 hours of incubation. The final concentration of vanillin in the medium was increased up to 3.5 mM after a 6-hour incubation by sequential spiking of 1.1 mM ferulic acid. The resting cells could be reused up to four times maintaining the production yield levels over 50%, thus increasing three times the vanillin obtained per gram of biomass. Ferulic acid can be efficiently converted to vanillin, without accumulation of undesirable vanillin reduction/oxidation products, using E. coli JM109 cells expressing genes from the ferulic

  4. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin.

    Fleige, Christian; Meyer, Florian; Steinbüchel, Alexander

    2016-06-01

    The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation resulted in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement was achieved with constitutive expression of the vanillin anabolism genes ech and fcs, coding for the enzymes feruloyl-coenzyme A (CoA) synthetase (fcs) and enoyl-CoA hydratase/aldolase (ech). The transcription of both genes was shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin was efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase was eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, was obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter was achieved, at the expense of a lower molar yield, by using an improved feeding strategy. This is the highest reported vanillin concentration reached in microbial fermentation processes without extraction of the product. Furthermore, the vanillin was produced almost without by-products, with a molar yield that nearly approached the theoretical maximum. Much effort has been put into optimization of the biotechnological production of natural vanillin. The demand for this compound is growing due to increased consumer concerns regarding chemically produced food additives. Since this compound is toxic to most

  5. Vanillin production using metabolically engineered Escherichia coli under non-growing conditions

    Fava Fabio

    2007-04-01

    Full Text Available Abstract Background Vanillin is one of the most important aromatic flavour compounds used in the food and cosmetic industries. Natural vanillin is extracted from vanilla beans and is relatively expensive. Moreover, the consumer demand for natural vanillin highly exceeds the amount of vanillin extracted by plant sources. This has led to the investigation of other routes to obtain this flavour such as the biotechnological production from ferulic acid. Studies concerning the use of engineered recombinant Escherichia coli cells as biocatalysts for vanillin production are described in the literature, but yield optimization and biotransformation conditions have not been investigated in details. Results Effect of plasmid copy number in metabolic engineering of E. coli for the synthesis of vanillin has been evaluated by the use of genes encoding feruloyl-CoA synthetase and feruloyl hydratase/aldolase from Pseudomonas fluorescens BF13. The higher vanillin production yield was obtained using resting cells of E. coli strain JM109 harbouring a low-copy number vector and a promoter exhibiting a low activity to drive the expression of the catabolic genes. Optimization of the bioconversion of ferulic acid to vanillin was accomplished by a response surface methodology. The experimental conditions that allowed us to obtain high values for response functions were 3.3 mM ferulic acid and 4.5 g/L of biomass, with a yield of 70.6% and specific productivity of 5.9 μmoles/g × min after 3 hours of incubation. The final concentration of vanillin in the medium was increased up to 3.5 mM after a 6-hour incubation by sequential spiking of 1.1 mM ferulic acid. The resting cells could be reused up to four times maintaining the production yield levels over 50%, thus increasing three times the vanillin obtained per gram of biomass. Conclusion Ferulic acid can be efficiently converted to vanillin, without accumulation of undesirable vanillin reduction/oxidation products

  6. Net community production and metabolic balance at the oligotrophic ocean site, station ALOHA

    le B. Williams, Peter J.; Morris, Paul J.; Karl, David M.

    2004-11-01

    To test the hypothesis that in oligotrophic areas of the ocean respiration exceeds production, a 12-month study was undertaken of in vitro-determined net oxygen production and consumption in the top 150 m of the water column at the extreme oligotrophic site, Station ALOHA, in the North Pacific subtropical gyre. Throughout the year the water column was observed to be in metabolic deficit, the calculated cumulative shortfall being 9±1.7 mol O2 m-2 a-1 (approximately 100 g C m-2 a-1), an amount equivalent to 40% of measured production (annual estimated rates of production and consumption were, respectively, 22 and 31 mol O2 m-2 a-1). We consider three possible explanations for the observed deficit: the in vitro oxygen rate measurements, in themselves, are fundamentally flawed and should be discounted, the observations are correct and the observed deficit is a true account of the balance of oxygen (and organic carbon) at Station ALOHA, or the observations are correct as they stand, but need not be interpreted as organic carbon imbalance for that ecosystem. We find no error unique to the oxygen rate measurements themselves. We find also no evidence that the associated organic carbon deficit can be sustained over the long-term by internal organic reserves or by external subsidy. Accordingly we accept the geochemical findings that calculated in situ oxygen flux requires the euphotic zone of the water column at this site to be slightly (circa 2 mol C m-2 a-1) autotrophic, in contrast to the simple analysis of our observations which gives a net heterotrophic water column. We discuss a number of processes that may give rise to the observed discrepancy. In part it may derive from the difficulty of reproducing the variations in the light field experienced by an algal cell due to vertical advection. It may also derive from the intermittency of production. This latter effect would manifest itself in the following manner. Because of its universal distribution in the food web

  7. Peasant territoriality in the South of Argentina. Productive and labour changes as forms of resistance

    Mónica Bendini

    2014-03-01

    Full Text Available In a context of territorial expansion of capital and social changes in rural life, the article addresses the issue of the persistence of peasant producers in northern Patagonia (Argentina and the development of various adaptive strategies as a form of resistance expulsion as producers. The changes are researched not only in households but also in the areas of life and work of those producers who call themselves "crianceros" in two regions of extensive livestock export oriented, wool and mohair hair. The cases show that the territorial expansion of capital involves social dynamics that go beyond productive and institutional transformations. Family strategies are more complex and also generate changes in other levels of rurality, in the mobility, in the habitat, in the activities of the countryside and towns, in the horizons of rural life. 

  8. The use of hexacyanoferrates in different forms to reduce radiocaesium contamination of animal products in Russia

    Ratnikov, A.N.; Vasiliev, A.V.; Alexakhin, R.M.; Krasnova, E.G.; Pasternak, A.D.; Howard, B.J.; Hove, K.; Strand, P.

    1998-01-01

    Hexacyanoferrates have been identified as highly effective radiocaesium binders which effectively reduce radiocaesium uptake and transfer to milk and meat. In Russia a hexacyanoferrate called ferrocyn has been produced for use as a countermeasure. In 1989-1992, experiments were undertaken in Russia to study the effectiveness of four different ferrocyn materials as 137 Cs binders, their potential toxicity, effect on production rates of cow milk, effect on animal health and ease of implementation in routine agricultural practice. Four different ferrocyn delivery forms have been used: 98% pure powder, sustained release rumen boli (15% ferrocyn), salt licks (10% ferrocyn) and sawdust with 10% ferrocyn adsorbed (bifege). In initial experiments with different cows, sheep and pigs these four ferrocyn materials were effective in reducing radiocaesium transfer to animal products. Daily administration of ferrocyn powder at a rate of 3-5 g per cow reduced 137 Cs transfer by up to 90% in milk. One single administration of three boli per cow (containing 30 g ferrocyn per boli) reduced 137 Cs transfer by 50-75% for a period of 2 months. Salt licks containing 10% ferrocyn (0.22 kg ferrocyn per 2.2 kg briquette provided once) reduced transfer of 137 Cs up to twofold for up to 10 days whilst bifege, given at a rate of 30-60 g day -1 (3-6 g day -1 ferrocyn), reduced 137 Cs transfer by 90-95%. However, large-scale application of these ferrocyn materials on collective and private farms in agricultural trials in 1994 resulted in a lower effectiveness. Therefore, in 1996 a comparative assessment of the application of the four ferrocyn forms was made under carefully controlled conditions. The results fully validated the previous experimental data, and showed the importance of meeting recommended procedures for treatment, particularly when hexacyanoferrates are administered on a day-to-day basis

  9. Process for the removal of acid forming gases from exhaust gases and production of phosphoric acid

    Chang, Shih-Ger; Liu, David K.

    1992-01-01

    Exhaust gases are treated to remove NO or NO.sub.x and SO.sub.2 by contacting the gases with an aqueous emulsion or suspension of yellow phosphorous preferably in a wet scrubber. The addition of yellow phosphorous in the system induces the production of O.sub.3 which subsequently oxidizes NO to NO.sub.2. The resulting NO.sub.2 dissolves readily and can be reduced to form ammonium ions by dissolved SO.sub.2 under appropriate conditions. In a 20 acfm system, yellow phosphorous is oxidized to yield P.sub.2 O.sub.5 which picks up water to form H.sub.3 PO.sub.4 mists and can be collected as a valuable product. The pressure is not critical, and ambient pressures are used. Hot water temperatures are best, but economics suggest about 50.degree. C. The amount of yellow phosphorus used will vary with the composition of the exhaust gas, less than 3% for small concentrations of NO, and 10% or higher for concentrations above say 1000 ppm. Similarly, the pH will vary with the composition being treated, and it is adjusted with a suitable alkali. For mixtures of NO.sub.x and SO.sub.2, alkalis that are used for flue gas desulfurization are preferred. With this process, better than 90% of SO.sub.2 and NO in simulated flue gas can be removed. Stoichiometric ratios (P/NO) ranging between 0.6 and 1.5 were obtained.

  10. Production of soybean isoflavone genistein in non-legume plants via genetically modified secondary metabolism pathway.

    Liu, Rongrong; Hu, Yuanlei; Li, Jialin; Lin, Zhongping

    2007-01-01

    Genetic modification of secondary metabolic pathways to produce desirable natural products is an attractive approach in plant biotechnology. In our study, we attempted to produce a typical soybean isoflavone genistein, a well-known health-promoting metabolite, in non-legume plants via genetic engineering. Both overexpression and antisense suppression strategies were used to manipulate the expression of several genes encoding key enzymes in the flavonoids/isoflavonoids pathway in transgenic tobacco, lettuce, and petunia. Introducing soybean isoflavone synthase (IFS) into these plants, which naturally do not produce isoflavonoids due to a lack of this leguminous enzyme, resulted in genistein biosynthesis in tobacco petals, petunia leaves and petals, and lettuce leaves. In tobacco, when flavanone 3-hydroxylase (F3H) expression was suppressed by its antisense gene while soybean IFS was overexpressed at the same time, genistein yield increased prominently. In addition, overexpression of phenylalanine ammonia-lyase (PAL) also led to an enhanced genistein production in tobacco petals and lettuce leaves in the presence of IFS than in the plants that overexpressed only IFS.

  11. Enhancing GDP-fucose production in recombinant Escherichia coli by metabolic pathway engineering.

    Zhai, Yafei; Han, Donglei; Pan, Ying; Wang, Shuaishuai; Fang, Junqiang; Wang, Peng; Liu, Xian-wei

    2015-02-01

    Guanosine 5'-diphosphate (GDP)-fucose is the indispensible donor substrate for fucosyltransferase-catalyzed synthesis of fucose-containing biomolecules, which have been found involving in various biological functions. In this work, the salvage pathway for GDP-fucose biosynthesis from Bacterioides fragilis was introduced into Escherichia coli. Besides, the biosynthesis of guanosine 5'-triphosphate (GTP), an essential substrate for GDP-fucose biosynthesis, was enhanced via overexpression of enzymes involved in the salvage pathway of GTP biosynthesis. The production capacities of metabolically engineered strains bearing different combinations of recombinant enzymes were compared. The shake flask fermentation of the strain expressing Fkp, Gpt, Gmk and Ndk obtained the maximum GDP-fucose content of 4.6 ± 0.22 μmol/g (dry cell mass), which is 4.2 fold that of the strain only expressing Fkp. Through fed-batch fermentation, the GDP-fucose content further rose to 6.6 ± 0.14 μmol/g (dry cell mass). In addition to a better productivity than previous fermentation processes based on the de novo pathway for GDP-fucose biosynthesis, the established schemes in this work also have the advantage to be a potential avenue to GDP-fucose analogs encompassing chemical modification on the fucose residue. Copyright © 2014 Elsevier Inc. All rights reserved.

  12. Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells

    Fariborz Bahrami

    2014-01-01

    Full Text Available Interleukin-8 (IL-8 plays key roles in both chronic inflammatory diseases and tumor modulation. We previously observed that IL-8 secretion and function can be modulated by nucleotide (P2 receptors. Here we investigated whether IL-8 release by intestinal epithelial HT-29 cells, a cancer cell line, is modulated by extracellular nucleotide metabolism. We first identified that HT-29 cells regulated adenosine and adenine nucleotide concentration at their surface by the expression of the ectoenzymes NTPDase2, ecto-5′-nucleotidase, and adenylate kinase. The expression of the ectoenzymes was evaluated by RT-PCR, qPCR, and immunoblotting, and their activity was analyzed by RP-HPLC of the products and by detection of Pi produced from the hydrolysis of ATP, ADP, and AMP. In response to poly (I:C, with or without ATP and/or ADP, HT-29 cells released IL-8 and this secretion was modulated by the presence of NTPDase2 and adenylate kinase. Taken together, these results demonstrate the presence of 3 ectoenzymes at the surface of HT-29 cells that control nucleotide levels and adenosine production (NTPDase2, ecto-5′-nucleotidase and adenylate kinase and that P2 receptor-mediated signaling controls IL-8 release in HT-29 cells which is modulated by the presence of NTPDase2 and adenylate kinase.

  13. Design of process parameters for the incremental tube forming (ITF) by FEM to control product properties

    Nazari, Esmaeil; Löbbe, Christian; Gallus, Stefan; Izadyar, S. Ahmad; Tekkaya, A. Erman

    2018-05-01

    The incremental tube forming (ITF) is a process combination of the kinematic tube bending and spinning to shape high strength and tailored tubes with variable diameters and thicknesses. In contrast to conventional bending methods, the compressive stress superposition by the spinning process facilitates low bending stresses, so that geometrical errors are avoided and the shape accuracy is improved. The study reveals the interaction of plastic strains of the rolling and bending process through an explicit FEM investigation. For this purpose, the three-dimensional machine set-up is discretized and modeled in terms of the fully disclosed spinning process during the gradual deflection of the tube end for bending. The analysis shows that, depending on the forming tool shape, the stress superposition is accompanied by high plastic strains. Furthermore, this phenomenon is explained by the three dimensional normal and shear strains during the incremental spinning. Analyzing the strains history also shows a nonlinearity between the strains by bending and spinning. It is also shown that process parameters like rotational velocity of the spinning rolls have a huge influence on the deformation pattern. Finally, the method is used for the manufacturing of an example product, which reveals the high process flexibility. In one clamp a component with a graded wall thickness and outside diameter along the longitudinal axis is produced.

  14. Biofilm-forming capacity in biogenic amine-producing bacteria isolated from dairy products.

    Maria eDiaz

    2016-05-01

    Full Text Available Biofilms on the surface of food industry equipment are reservoirs of potentially food-contaminating bacteria - both spoilage and pathogenic. However, the capacity of biogenic amine (BA-producers to form biofilms has remained largely unexamined. BAs are low molecular weight, biologically active compounds that in food can reach concentrations high enough to be a toxicological hazard. Fermented foods, especially some types of cheese, accumulate the highest BA concentrations of all. The present work examines the biofilm-forming capacity of 56 BA-producing strains belonging to three genera and 10 species (12 Enterococcus faecalis, 6 Enterococcus faecium, 6 Enterococcus durans, 1 Enterococcus hirae, 12 Lactococcus lactis, 7 Lactobacillus vaginalis, 2 Lactobacillus curvatus, 2 Lactobacillus brevis, 1 Lactobacillus reuteri and 7 Lactobacillus parabuchneri, all isolated from dairy products. Strains of all the tested species - except for L. vaginalis - were able to produce biofilms on polystyrene and adhered to stainless steel. However, the biomass produced in biofilms was strain-dependent. These results suggest that biofilms may provide a route via which fermented foods can become contaminated by BA-producing microorganisms.

  15. Matrix production and organization by endothelial colony forming cells in mechanically strained engineered tissue constructs.

    Nicky de Jonge

    Full Text Available AIMS: Tissue engineering is an innovative method to restore cardiovascular tissue function by implanting either an in vitro cultured tissue or a degradable, mechanically functional scaffold that gradually transforms into a living neo-tissue by recruiting tissue forming cells at the site of implantation. Circulating endothelial colony forming cells (ECFCs are capable of differentiating into endothelial cells as well as a mesenchymal ECM-producing phenotype, undergoing Endothelial-to-Mesenchymal-transition (EndoMT. We investigated the potential of ECFCs to produce and organize ECM under the influence of static and cyclic mechanical strain, as well as stimulation with transforming growth factor β1 (TGFβ1. METHODS AND RESULTS: A fibrin-based 3D tissue model was used to simulate neo-tissue formation. Extracellular matrix organization was monitored using confocal laser-scanning microscopy. ECFCs produced collagen and also elastin, but did not form an organized matrix, except when cultured with TGFβ1 under static strain. Here, collagen was aligned more parallel to the strain direction, similar to Human Vena Saphena Cell-seeded controls. Priming ECFC with TGFβ1 before exposing them to strain led to more homogenous matrix production. CONCLUSIONS: Biochemical and mechanical cues can induce extracellular matrix formation by ECFCs in tissue models that mimic early tissue formation. Our findings suggest that priming with bioactives may be required to optimize neo-tissue development with ECFCs and has important consequences for the timing of stimuli applied to scaffold designs for both in vitro and in situ cardiovascular tissue engineering. The results obtained with ECFCs differ from those obtained with other cell sources, such as vena saphena-derived myofibroblasts, underlining the need for experimental models like ours to test novel cell sources for cardiovascular tissue engineering.

  16. Capsule Production and Glucose Metabolism Dictate Fitness during Serratia marcescens Bacteremia.

    Anderson, Mark T; Mitchell, Lindsay A; Zhao, Lili; Mobley, Harry L T

    2017-05-23

    Serratia marcescens is an opportunistic pathogen that causes a range of human infections, including bacteremia, keratitis, wound infections, and urinary tract infections. Compared to other members of the Enterobacteriaceae family, the genetic factors that facilitate Serratia proliferation within the mammalian host are less well defined. An in vivo screen of transposon insertion mutants identified 212 S. marcescens fitness genes that contribute to bacterial survival in a murine model of bloodstream infection. Among those identified, 11 genes were located within an 18-gene cluster encoding predicted extracellular polysaccharide biosynthesis proteins. A mutation in the wzx gene contained within this locus conferred a loss of fitness in competition infections with the wild-type strain and a reduction in extracellular uronic acids correlating with capsule loss. A second gene, pgm , encoding a phosphoglucomutase exhibited similar capsule-deficient phenotypes, linking central glucose metabolism with capsule production and fitness of Serratia during mammalian infection. Further evidence of the importance of central metabolism was obtained with a pfkA glycolytic mutant that demonstrated reduced replication in human serum and during murine infection. An MgtB magnesium transporter homolog was also among the fitness factors identified, and an S. marcescens mgtB mutant exhibited decreased growth in defined medium containing low concentrations of magnesium and was outcompeted ~10-fold by wild-type bacteria in mice. Together, these newly identified genes provide a more complete understanding of the specific requirements for S. marcescens survival in the mammalian host and provide a framework for further investigation of the means by which S. marcescens causes opportunistic infections. IMPORTANCE Serratia marcescens is a remarkably prolific organism that replicates in diverse environments, including as an opportunistic pathogen in human bacteremia. The genetic requirements for

  17. Genetic parameters of product quality and hepatic metabolism in fattened mule ducks.

    Marie-Etancelin, C; Basso, B; Davail, S; Gontier, K; Fernandez, X; Vitezica, Z G; Bastianelli, D; Baéza, E; Bernadet, M-D; Guy, G; Brun, J-M; Legarra, A

    2011-03-01

    Genetic parameters of traits related to hepatic lipid metabolism, carcass composition, and product quality of overfed mule ducks were estimated on both parental lines of this hybrid: the common duck line for the maternal side and the Muscovy line for the paternal side. The originality of the statistical model was to include simultaneously the additive genetic effect of the common ducks and that of the Muscovy ducks, revealing a greater genetic determinism in common than in Muscovy. Plasma metabolic indicators (glucose, triglyceride, and cholesterol contents) were heritable, in particular at the end of the overfeeding period, and heritabilities increased with the overfeeding stage. Carcass composition traits were highly heritable in the common line, with values ranging from 0.15 for liver weight, 0.21 for carcass weight, and 0.25 for abdominal fat weight to 0.32 for breast muscle weight. Heritabilities of technological outputs were greater for the fatty liver (0.19 and 0.08, respectively, on common and Muscovy sides for liver melting rate) than for the pectoralis major muscle (between 0.02 and 0.05 on both parental sides for cooking losses). Fortunately, the processing industry is mainly facing problems in liver quality, such as too high of a melting rate, than in meat quality. The meat quality appraisal criteria (such as texture and cooking losses), usually dependent on pH and the rate of decline of pH, were also very lowly heritable. This study demonstrated that genetic determinism of meat quality and ability of overfeeding is not similar in the common population and in the Muscovy population; traits related to fattening, muscle development, and BW have heritability values from 2 to 4 times greater on the common line than on the Muscovy line, which is relevant for considering different selection strategies.

  18. Capsule Production and Glucose Metabolism Dictate Fitness during Serratia marcescens Bacteremia

    Mark T. Anderson

    2017-05-01

    Full Text Available Serratia marcescens is an opportunistic pathogen that causes a range of human infections, including bacteremia, keratitis, wound infections, and urinary tract infections. Compared to other members of the Enterobacteriaceae family, the genetic factors that facilitate Serratia proliferation within the mammalian host are less well defined. An in vivo screen of transposon insertion mutants identified 212 S. marcescens fitness genes that contribute to bacterial survival in a murine model of bloodstream infection. Among those identified, 11 genes were located within an 18-gene cluster encoding predicted extracellular polysaccharide biosynthesis proteins. A mutation in the wzx gene contained within this locus conferred a loss of fitness in competition infections with the wild-type strain and a reduction in extracellular uronic acids correlating with capsule loss. A second gene, pgm, encoding a phosphoglucomutase exhibited similar capsule-deficient phenotypes, linking central glucose metabolism with capsule production and fitness of Serratia during mammalian infection. Further evidence of the importance of central metabolism was obtained with a pfkA glycolytic mutant that demonstrated reduced replication in human serum and during murine infection. An MgtB magnesium transporter homolog was also among the fitness factors identified, and an S. marcescens mgtB mutant exhibited decreased growth in defined medium containing low concentrations of magnesium and was outcompeted ~10-fold by wild-type bacteria in mice. Together, these newly identified genes provide a more complete understanding of the specific requirements for S. marcescens survival in the mammalian host and provide a framework for further investigation of the means by which S. marcescens causes opportunistic infections.

  19. The Essential Role of Cholesterol Metabolism in the Intracellular Survival of Mycobacterium leprae Is Not Coupled to Central Carbon Metabolism and Energy Production.

    Marques, Maria Angela M; Berrêdo-Pinho, Marcia; Rosa, Thabatta L S A; Pujari, Venugopal; Lemes, Robertha M R; Lery, Leticia M S; Silva, Carlos Adriano M; Guimarães, Ana Carolina R; Atella, Georgia C; Wheat, William H; Brennan, Patrick J; Crick, Dean C; Belisle, John T; Pessolani, Maria Cristina V

    2015-12-01

    Mycobacterium leprae induces the formation of lipid droplets, which are recruited to pathogen-containing phagosomes in infected macrophages and Schwann cells. Cholesterol is among the lipids with increased abundance in M. leprae-infected cells, and intracellular survival relies on cholesterol accumulation. The present study investigated the capacity of M. leprae to acquire and metabolize cholesterol. In silico analyses showed that oxidation of cholesterol to cholest-4-en-3-one (cholestenone), the first step of cholesterol degradation catalyzed by the enzyme 3β-hydroxysteroid dehydrogenase (3β-HSD), is apparently the only portion of the cholesterol catabolic pathway seen in Mycobacterium tuberculosis preserved by M. leprae. Incubation of bacteria with radiolabeled cholesterol confirmed the in silico predictions. Radiorespirometry and lipid analyses performed after incubating M. leprae with [4-(14)C]cholesterol or [26-(14)C]cholesterol showed the inability of this pathogen to metabolize the sterol rings or the side chain of cholesterol as a source of energy and carbon. However, the bacteria avidly incorporated cholesterol and, as expected, converted it to cholestenone both in vitro and in vivo. Our data indicate that M. leprae has lost the capacity to degrade and utilize cholesterol as a nutritional source but retains the enzyme responsible for its oxidation to cholestenone. Thus, the essential role of cholesterol metabolism in the intracellular survival of M. leprae is uncoupled from central carbon metabolism and energy production. Further elucidation of cholesterol metabolism in the host cell during M. leprae infection will establish the mechanism by which this lipid supports M. leprae intracellular survival and will open new avenues for novel leprosy therapies. Our study focused on the obligate intracellular pathogen Mycobacterium leprae and its capacity to metabolize cholesterol. The data make an important contribution for those interested in understanding the

  20. Enhanced volatile fatty acids production from anaerobic fermentation of food waste: A mini-review focusing on acidogenic metabolic pathways.

    Zhou, Miaomiao; Yan, Binghua; Wong, Jonathan W C; Zhang, Yang

    2018-01-01

    Recently, efficient disposal of food waste (FW) with potential resource recovery has attracted great attentions. Due to its easily biodegradable nature, rich nutrient availability and high moisture content, FW is regarded as favorable substrate for anaerobic digestion (AD). Both waste disposal and energy recovery can be fulfilled during AD of FW. Volatile fatty acids (VFAs) which are the products of the first-two stages of AD, are widely applied in chemical industry as platform chemicals recently. Concentration and distribution of VFAs is the result of acidogenic metabolic pathways, which can be affected by the micro-environment (e.g. pH) in the digester. Hence, the clear elucidation of the acidogenic metabolic pathways is essential for optimization of acidogenic process for efficient product recovery. This review summarizes major acidogenic metabolic pathways and regulating strategies for enhancing VFAs recovery during acidogenic fermentation of FW. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Abnormal iron metabolism and oxidative stress in mice expressing a mutant form of the ferritin light polypeptide gene

    Barbeito, Ana G.; Garringer, Holly J.; Baraibar, Martin A.; Gao, Xiaoying; Arredondo, Miguel; Núñez, Marco T.; Smith, Mark A.; Ghetti, Bernardino; Vidal, Ruben

    2009-01-01

    Insertional mutations in exon 4 of the ferritin light chain (FTL) gene are associated with hereditary ferritinopathy (HF) or neuroferritinopathy, an autosomal dominant neurodegenerative disease characterized by progressive impairment of motor and cognitive functions. To determine the pathogenic mechanisms by which mutations in FTL lead to neurodegeneration, we investigated iron metabolism and markers of oxidative stress in the brain of transgenic (Tg) mice that express the mutant human FTL498-499InsTC cDNA. Compared with wild-type mice, brain extracts from Tg (FTL-Tg) mice showed an increase in the cytoplasmic levels of both FTL and ferritin heavy chain polypeptides, a decrease in the protein and mRNA levels of transferrin receptor-1, and a significant increase in iron levels. Transgenic mice also showed the presence of markers for lipid peroxidation, protein carbonyls, and nitrone–protein adducts in the brain. However, gene expression analysis of iron management proteins in the liver of Tg mice indicates that the FTL-Tg mouse liver is iron deficient. Our data suggest that disruption of iron metabolism in the brain has a primary role in the process of neurodegeneration in HF and that the pathogenesis of HF is likely to result from a combination of reduction in iron storage function and enhanced toxicity associated with iron-induced ferritin aggregates in the brain. PMID:19519778

  2. A unified form of exact-MSR codes via product-matrix frameworks

    Lin, Sian Jheng

    2015-02-01

    Regenerating codes represent a class of block codes applicable for distributed storage systems. The [n, k, d] regenerating code has data recovery capability while possessing arbitrary k out of n code fragments, and supports the capability for code fragment regeneration through the use of other arbitrary d fragments, for k ≤ d ≤ n - 1. Minimum storage regenerating (MSR) codes are a subset of regenerating codes containing the minimal size of each code fragment. The first explicit construction of MSR codes that can perform exact regeneration (named exact-MSR codes) for d ≥ 2k - 2 has been presented via a product-matrix framework. This paper addresses some of the practical issues on the construction of exact-MSR codes. The major contributions of this paper include as follows. A new product-matrix framework is proposed to directly include all feasible exact-MSR codes for d ≥ 2k - 2. The mechanism for a systematic version of exact-MSR code is proposed to minimize the computational complexities for the process of message-symbol remapping. Two practical forms of encoding matrices are presented to reduce the size of the finite field.

  3. A unified form of exact-MSR codes via product-matrix frameworks

    Lin, Sian Jheng; Chung, Weiho; Han, Yunghsiangsam; Al-Naffouri, Tareq Y.

    2015-01-01

    Regenerating codes represent a class of block codes applicable for distributed storage systems. The [n, k, d] regenerating code has data recovery capability while possessing arbitrary k out of n code fragments, and supports the capability for code fragment regeneration through the use of other arbitrary d fragments, for k ≤ d ≤ n - 1. Minimum storage regenerating (MSR) codes are a subset of regenerating codes containing the minimal size of each code fragment. The first explicit construction of MSR codes that can perform exact regeneration (named exact-MSR codes) for d ≥ 2k - 2 has been presented via a product-matrix framework. This paper addresses some of the practical issues on the construction of exact-MSR codes. The major contributions of this paper include as follows. A new product-matrix framework is proposed to directly include all feasible exact-MSR codes for d ≥ 2k - 2. The mechanism for a systematic version of exact-MSR code is proposed to minimize the computational complexities for the process of message-symbol remapping. Two practical forms of encoding matrices are presented to reduce the size of the finite field.

  4. Effect of mustard gas hydrolysis products on the development of water-bloom forming cyanobacteria

    Zaytseva Tatyana

    2017-03-01

    Full Text Available Mustard gas and its hydrolysis products (MGHP belong to stable organochlorine compounds with high toxicity and broad spectrum of activity. Since the Second World War many aquatic ecosystems including the Baltic and the Adriatic Sea as well as the coastal waters of Japan, the USA, the UK, Australia have been contaminated with mustard gas due to the dumping of chemical weapon. Mustard gas and its hydrolysis products have a negative impact on aquatic life including microbiota. The aim of this work was to define the effect of MGHP on the growth, photosynthetic activity and synthesis of secondary metabolites by water-bloom forming cyanobacteria Trichormus variabilis, Aphanizomenon flos-aquae, Microcystis aeruginosa, Nodularia spumigena. Microbiological, chromatographic, spectrophotometric methods were used. The growth inhibition test with MGHP on cyanobacteria showed influence on the concentration EC50 within the range of 5.5 – 11.2 mg of organochlorine compounds (ОCC per liter. The synthesis of chlorophyll a was also decreased. It was shown that the chlorophyll synthesis was more sensitive to MGHP than the growth of cyanobacteria. NGHP induced enhanced excretion of exopolysaccharides. Low concentration of MGHP – 0.3 mg OCC/l - promoted the growth of toxigenic cyanobacterium Microcystis aeruginosa and increased microcystin-LR concentration in the environment. enhanced excretion of such metabolites as polysaccharides and cyanotoxins has a serious negative impact on water pollution due to MGHP.

  5. A comparative study on the raft chemical properties of various alginate antacid raft-forming products.

    Dettmar, Peter W; Gil-Gonzalez, Diana; Fisher, Jeanine; Flint, Lucy; Rainforth, Daniel; Moreno-Herrera, Antonio; Potts, Mark

    2018-01-01

    Research to measure the chemical characterization of alginate rafts for good raft performance and ascertain how formulation can affect chemical parameters. A selection of alginate formulations was investigated all claiming to be proficient raft formers with significance between products established and ranked. Procedures were selected which demonstrated the chemical characterization allowing rafts to effectively impede the reflux into the esophagus or in severe cases to be refluxed preferentially into the esophagus and exert a demulcent effect, with focus of current research on methods which complement previous studies centered on physical properties. The alginate content was analyzed by a newly developed HPLC method. Methods were used to determine the neutralization profile and the acid neutralization within the raft determined along with how raft structure affects neutralization. Alginate content of Gaviscon Double Action (GDA) within the raft was significantly superior (p raft acid neutralization capacity were GDA and Rennie Duo, the latter product not being a raft former. Raft structure was key and GDA had the right level of porosity to allow for longer duration of neutralization. Alginate formulations require three chemical reactions to take place simultaneously: transformation to alginic acid, sodium carbonate reacting to form carbon dioxide, calcium releasing free calcium ions to bind with alginic acid providing strength to raft formation. GDA was significantly superior (p <.0001) to all other comparators.

  6. Filterability of corrosion products formed between carbon steel and water. Influence of temperature and oxygen content

    Kelen, T.; Falk, I.

    1975-09-01

    A laboratory investigation has been made for the purpose of studying the influence of temperature and oxygen content on the filterability of corrosion products formed between carbon-steel and water. The experiments were performed in a high temperature loop where the water is initially heated in a pre-heater, then cooled and finally filtered. The corrosion products were transferred to thewater from a carbon-steel surface that had previously been neutron activated and the amount of iron present was determined from measurements of the γ-radiation emitted by Fe-59. Filterability was then computed as the ratio between the total amount of iron in the water phase and the amount of iron retained on the filter. The investigation covers a series of experiments at filtering temperatures of 20, 90 and 160 dec G, pre-heater temperatures up to 300 deg C and oxygen contents of 10 and 300 ppb O 2 . In addition the extent of iron deposition in the pre-heater and heat regulator has been determined after each series of experiments. Filterability exhibited a pronounced dependence upon both the filter and pre-heater temperatures and also upon the oxygen content. Among the conclusions to which the results lead is the observation that a strict comparison of filterability values for the fraction of corrosion products in cooled water samples is impossible when these are taken from 1) different sections of a high temperature system 2) a single sampling point while the system is being run up 3) two separate systems (e.g. steam boilers) operated at different temperatures 4) two separate systems operated at different oxygen contents. It accordingly appears advizable to restrict the use of cold-filtered samples from conventional steam-raising plants to the comparison of values relating to a single sampling point under constant operating conditions. (author)

  7. Multi-omic profiling of EPO-producing CHO cell panel reveals metabolic adaptation to heterologous protein production

    Ley, Daniel; Kazemi Seresht, Ali; Engmark, Mikael

    The Chinese hamster ovary (CHO) cell line is the predominant mammalian cell factory for production of therapeutic glycoproteins. In this work, we aimed to study bottlenecks in the secretory pathway associated with the production of human erythropoietin (EPO) in CHO cells. In connection to this, we...... discovered indications of metabolic adaptation of the amino acid catabolism in favor of heterologous protein production. We established a panel of stably EPO expressing CHO-K1 clones spanning a 25-fold productivity range and characterized the clones in batch and chemostat cultures. For this, we employed...... a multi-omic physiological characterization including metabolic foot printing of amino acids, metabolite fingerprinting of glycolytic intermediates, NAD(P)H-/NAD(P)+ and adenosine nucleotide phosphates. We used qPCR, qRT-PCR, western blots and Affymetrix CHO microarrays to assess EPO gene copy numbers...

  8. Water deprivation induces appetite and alters metabolic strategy in Notomys alexis: unique mechanisms for water production in the desert.

    Takei, Yoshio; Bartolo, Ray C; Fujihara, Hiroaki; Ueta, Yoichi; Donald, John A

    2012-07-07

    Like many desert animals, the spinifex hopping mouse, Notomys alexis, can maintain water balance without drinking water. The role of the kidney in producing a small volume of highly concentrated urine has been well-documented, but little is known about the physiological mechanisms underpinning the metabolic production of water to offset obligatory water loss. In Notomys, we found that water deprivation (WD) induced a sustained high food intake that exceeded the pre-deprivation level, which was driven by parallel changes in plasma leptin and ghrelin and the expression of orexigenic and anorectic neuropeptide genes in the hypothalamus; these changed in a direction that would stimulate appetite. As the period of WD was prolonged, body fat disappeared but body mass increased gradually, which was attributed to hepatic glycogen storage. Switching metabolic strategy from lipids to carbohydrates would enhance metabolic water production per oxygen molecule, thus providing a mechanism to minimize respiratory water loss. The changes observed in appetite control and metabolic strategy in Notomys were absent or less prominent in laboratory mice. This study reveals novel mechanisms for appetite regulation and energy metabolism that could be essential for desert rodents to survive in xeric environments.

  9. Metabolism of fission products. I. The metabolism of the radioactive ashes obtained from the No. 5 Fukuryu Maru

    Kikuchi, T; Wakisaky, G; Kono, T; Hiroshi, G; Akagi, H; Yamamasu, T; Sugawa, I

    1954-01-01

    When the radioactive ashes were administered by mouth, the radioisotopes which were chiefly absorbed were alkaline earths, and were deposited mainly in the bones. When, after the removal of the alkaline earths, the radioisotopes contained in the radioactive ashes were administered by mouth in the form of chloride or citrate, the radioisotopes chiefly absorbed were heavy metals such as Ru and Rh.

  10. Formation of seed productivity in lettuce Lactuca sativa var. secalina L. depending on the form of inflorescence habitus

    Н. В. Лещук

    2009-12-01

    Full Text Available It is explained the essence of the formation of seed production of lettuce Lactuca sativa var. secalina L. depending on the form of the inflorescence habitus, which formed during the different areas of plant nutrition. It is substantiated the mechanism of seed formation on different parts of the inflorescence of female plant with identification of it’s sowing quality. It is developed the agrotehnologcal measures aimed at improving the productivity of lettuce seed by overcoming its matrical heteroquality

  11. Formation of seed productivity in lettuce Lactuca sativa var. secalina L. depending on the form of inflorescence habitus

    Н. В. Лещук

    2009-01-01

    It is explained the essence of the formation of seed production of lettuce Lactuca sativa var. secalina L. depending on the form of the inflorescence habitus, which formed during the different areas of plant nutrition. It is substantiated the mechanism of seed formation on different parts of the inflorescence of female plant with identification of it’s sowing quality. It is developed the agrotehnologcal measures aimed at improving the productivity of lettuce seed by overcoming its matrical he...

  12. Renewable energy from Cyanobacteria: energy production optimization by metabolic pathway engineering.

    Quintana, Naira; Van der Kooy, Frank; Van de Rhee, Miranda D; Voshol, Gerben P; Verpoorte, Robert

    2011-08-01

    The need to develop and improve sustainable energy resources is of eminent importance due to the finite nature of our fossil fuels. This review paper deals with a third generation renewable energy resource which does not compete with our food resources, cyanobacteria. We discuss the current state of the art in developing different types of bioenergy (ethanol, biodiesel, hydrogen, etc.) from cyanobacteria. The major important biochemical pathways in cyanobacteria are highlighted, and the possibility to influence these pathways to improve the production of specific types of energy forms the major part of this review.

  13. Characterization of degradation products of amorphous and polymorphic forms of clopidogrel bisulphate under solid state stress conditions

    Raijada, Dhara K; Prasad, Bhagwat; Paudel, Amrit

    2010-01-01

    The present study deals with the stress degradation studies on amorphous and polymorphic forms of clopidogrel bisulphate. The objective was to characterize the degradation products and postulate mechanism of decomposition of the drug under solid state stress conditions. For that, amorphous form, ...

  14. Metabolic engineering of Escherichia coli to enhance shikimic acid production from sorbitol.

    Liu, Xianglei; Lin, Jun; Hu, Haifeng; Zhou, Bin; Zhu, Baoquan

    2014-09-01

    Shikimic acid (SA) is the key synthetic material of Oseltamivir, which is an effective drug for the prevention and treatment of influenza. In this study, to block the downstream metabolic pathway of SA, the shikimate kinase isoenzyme genes aroK and aroL were deleted by Red recombination. Moreover, the key enzyme genes aroG, aroB, tktA and aroE of SA pathway were co-expressed by constructing the recombinant vector pETDuet-GBAE. As a result, SA production of E. coli BW25113 (∆aroL/aroK, DE3)/pETDuet-GBAE reached 1,077.6 mg/l when low amounts of sorbitol (5 g/l) were fed in shake flasks. The yield was 3.7 times that when glucose was used (P sorbitol was an optimized carbon source for the high efficient accumulation of SA for the first time, which was applicable to use in the industry for high yields and low consumption.

  15. Exposure to gemfibrozil and atorvastatin affects cholesterol metabolism and steroid production in zebrafish (Danio rerio).

    Al-Habsi, Aziz A; Massarsky, Andrey; Moon, Thomas W

    2016-09-01

    The commonly used lipid-lowering pharmaceuticals gemfibrozil (GEM) and atorvastatin (ATV) are detected in the aquatic environment; however, their potential effects on non-target fish species are yet to be fully understood. This study examined the effects of GEM and/or ATV on female and male adult zebrafish after a 30d dietary exposure. The exposure led to changes in several biochemical parameters, including reduction in cholesterol, triglycerides, cortisol, testosterone, and estradiol. Changes in cholesterol and triglycerides were also associated with changes in transcript levels of key genes involved with cholesterol and lipid regulation, including SREBP2, HMGCR1, PPARα, and SREBP1. We also noted higher CYP3A65 and atrogin1 mRNA levels in drug-treated male fish. Sex differences were apparent in some of the examined parameters at both biochemical and molecular levels. This study supports these drugs affecting cholesterol metabolism and steroid production in adult zebrafish. We conclude that the reduction in cortisol may impair the ability of these fish to mount a suitable stress response, whereas the reduction of sex steroids may negatively affect reproduction. Copyright © 2015 Elsevier Inc. All rights reserved.

  16. Na+/K+-ATPase interaction with methylglyoxal as reactive metabolic side product.

    Svrckova, Marika; Zatloukalova, Martina; Dvorakova, Petra; Coufalova, Dominika; Novak, David; Hernychova, Lenka; Vacek, Jan

    2017-07-01

    Proteins are subject to oxidative modification and the formation of adducts with a broad spectrum of reactive species via enzymatic and non-enzymatic mechanisms. Here we report that in vitro non-enzymatic methylglyoxal (MGO) binding causes the inhibition and formation of MGO advanced glycation end-products (MAGEs) in Na + /K + -ATPase (NKA). Concretely, MGO adducts with NKA amino acid residues (mainly Arg) and N ε -(carboxymethyl)lysine (CML) formation were found. MGO is not only an inhibitor for solubilized NKA (IC 50 =91±16μM), but also for reconstituted NKA in the lipid bilayer environment, which was clearly demonstrated using a DPPC/DPPE liposome model in the presence or absence of the NKA-selective inhibitor ouabain. High-resolution mass spectrometric analysis of a tryptic digest of NKA isolated from pig (Sus scrofa) kidney indicates that the intracellular α-subunit is naturally (post-translationally) modified by MGO in vivo. In contrast to this, the β-subunit could only be modified by MGO artificially, and the transmembrane part of the protein did not undergo MGO binding under the experimental setup used. As with bovine serum albumin, serving as the water-soluble model, we also demonstrated a high binding capacity of MGO to water-poorly soluble NKA using a multi-spectral methodology based on electroanalytical, immunochemical and fluorimetric tools. In addition, a partial suppression of the MGO-mediated inhibitory effect could be observed in the presence of aminoguanidine (pimagedine), a glycation suppressor and MGO-scavenger. All the results here were obtained with the X-ray structure of NKA in the E1 conformation (3WGV) and could be used in the further interpretation of the functionality of this key enzyme in the presence of highly-reactive metabolic side-products, glycation agents and generally under oxidative stress conditions. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. Natural products, an important resource for discovery of multitarget drugs and functional food for regulation of hepatic glucose metabolism.

    Li, Jian; Yu, Haiyang; Wang, Sijian; Wang, Wei; Chen, Qian; Ma, Yanmin; Zhang, Yi; Wang, Tao

    2018-01-01

    Imbalanced hepatic glucose homeostasis is one of the critical pathologic events in the development of metabolic syndromes (MSs). Therefore, regulation of imbalanced hepatic glucose homeostasis is important in drug development for MS treatment. In this review, we discuss the major targets that regulate hepatic glucose homeostasis in human physiologic and pathophysiologic processes, involving hepatic glucose uptake, glycolysis and glycogen synthesis, and summarize their changes in MSs. Recent literature suggests the necessity of multitarget drugs in the management of MS disorder for regulation of imbalanced glucose homeostasis in both experimental models and MS patients. Here, we highlight the potential bioactive compounds from natural products with medicinal or health care values, and focus on polypharmacologic and multitarget natural products with effects on various signaling pathways in hepatic glucose metabolism. This review shows the advantage and feasibility of discovering multicompound-multitarget drugs from natural products, and providing a new perspective of ways on drug and functional food development for MSs.

  18. Net primary production of forest-forming species in climatic gradients of Eurasia

    V. A. Usoltsev

    2018-04-01

    Full Text Available When using biomass and net primary production (NPP databases compiled by the authors for 6 forest-forming species in a number of 6694 and 2192 sample plots correspondingly, a system of regression models of their NPP is designed and some species-specific regularities of NPP distribution in two climatic gradients (natural zonality and climate continentality are stated. It is found that according to a zonal gradient, aboveground and total NPP in 2-needled pine and spruce-fir forests are monotonically increasing in the direction from the northern to the southern tip of the continent, while larch and birch have the maximum in the southern moderate, and aspen and poplar – in the northern moderate zone, but oak forests do not show any significant pattern. Within a single zonal belt, the aboveground and total NPP of coniferous and deciduous are monotonically decreasing in direction from the Atlantic and Pacific coasts to the continentality pole in Yakutia. The understory NPP of all the species, except oak, monotonically increase towards the subequatorial zone. For oak forests, any clear regularity is not revealed. Within a single zonal belt, when approaching continentality pole, Pinus and Quercus NPP monotonically decreases and in other species, increases. Species-specific patterns in changing the relative indices of NPP (forest stand underground NPP to aboveground one and forest understory NPP to total forest stand one in gradients of the natural zonality and climate continentality are established.

  19. Use of Emanation Thermal Analysis in the characterization of nuclear waste forms and their alteration products

    Balek, V; Malek, Z.; Banba, T.; Mitamura, H.; Vance, E.R.

    1999-01-01

    Emanation Thermal Analysis (ETA) was used for the characterization of thermal behavior of two nuclear waste glasses, basalt volcanic glass and perovskite ceramics before and after hydrolytic treatment. The release of radon, formed by the spontaneous α-decay of 228 Th and 224 Ra and incorporated into samples to a maximum depth of 100 nm from the surface due to the recoil, was measured during heating of the samples from 20 to 1200degC and subsequent cooling. Temperatures of the annealing of surface roughness, micro-cracks and other defects, produced by manufacture and/or by subsequent treatment of glass and ceramic samples, were determined using the ETA. Microstructure changes of glass corrosion accompanying their dehydration and thermal decomposition were characterized by the radon release rate changes. The effect of hydrolytic alteration on the thermal behavior of the nuclear waste glass was revealed by ETA in an early corrosion stage. In the alteration product of the perovskite ceramics the diffusion mobility of radon was assessed in the temperature range 1000-1200degC. The thermal stability of radiation-induced defects in perovskite ceramic powder bombarded by He + ions to doses of 10 14 and 10 16 ions/cm 2 was determined by means of ETA. (author)

  20. 25 CFR 309.22 - What are examples of painting and other fine art forms that are Indian products?

    2010-04-01

    ... 25 Indians 2 2010-04-01 2010-04-01 false What are examples of painting and other fine art forms that are Indian products? 309.22 Section 309.22 Indians INDIAN ARTS AND CRAFTS BOARD, DEPARTMENT OF THE INTERIOR PROTECTION OF INDIAN ARTS AND CRAFTS PRODUCTS § 309.22 What are examples of painting and other...

  1. Diagnostic value of daily fluctuations in the free form of testosterone and cortisol in men with obesity and metabolic syndrome under the age of 50 years

    E. A. Kuznetsova

    2016-01-01

    Full Text Available The important pathogenetic link of obesity and metabolic syndrome (MS in men are disorders of testosterone production in the testes and adrenal adrenal cortisol production. Тestosterone deficiency and functional hypercortisolism have a mutual influence on each other. Comprehensive assessment of testosterone and cortisol balance and identify their daily variability in the saliva may improve the diagnosis of hormonal disorders in men with obesity and MS. In the present study, by comparative and correlation analysis assessed the circadian rhythm of cortisol and testosterone production in young and middle age men, finding the relationship between the concentration of these hormones in the morning and evening portions of saliva with the changes of anthropometric, hemodynamic and metabolic parameters. The study involved 35 patients with MS, 16 – with overweight and obesity without the MS and 19 – with normal body weight. In men with obesity and MS found a violation of the circadian rhythm of testosterone production. Evening saliva testosterone showed a close correlation with the parameters of the MSome and clinical symptoms of androgen deficiency, as well as a more pronounced decrease with age. Circadian rhythm of cortisol production was not violated, but cortisol was significantly increased in the evening portion of saliva in patients with obesity and MS. MS was associated not only with an increase in the concentration of free salivary cortisol in the evening hours, but with lower cortisol levels in the batch of saliva collected in the morning, which is consistent with other studies.

  2. Development of a data acquisition system in form of an apparatus which is specially designed for the study of the cerebral metabolism employing radioactive tracers

    Ponchon, J.

    1988-03-01

    The subject of this thesis is the development of a data acquisition system in form of an apparatus which is specially designed for the study of cerebral metabolism employing radioactive tracers. Currently, the test is based on the continuous inhalation of gas identified with oxygen 15. After an inhalation period of 6 to 8 minutes the local brain radioactivity attains an equilibrium which corresponds to the parameters of brain metabolism. The constant measurement of the radioactivity facilitates the monitoring of the evolution of these parameters. The apparatus consists of an assemby of lead collimators in which are lodged 26 gamma ray detectors (INa probes), each one of which scans a particular brain region. For ease of use and the provision of numerous possibilities, each detector has been assigned a multichannel pulse height analyser which has been developed by ourselves in order to correspond to and to comply with the demands of our tests. They are equipped with a device which automatically corrects for dead time thereby permitting the analysis of random impulses of a rate greater than 50.000 per second with a loss of less than 1%. The multichannel analysers are connected to a central computer which by means of suitably adapted software allows the calibration of different channel of measurement, controls, the choice of the zone of interest to be integrated and the presentation of integrated results in form of graphs [fr

  3. In-silico-driven metabolic engineering of Pseudomonas putida for enhanced production of poly-hydroxyalkanoates

    Poblete-Castro, I.; Binger, D.; Rodrigues, A.; Becker, J.; Martins Dos Santos, V.A.P.; Wittmann, C.

    2013-01-01

    Here, we present systems metabolic engineering driven by in-silico modeling to tailor Pseudomonas putida for synthesis of medium chain length PHAs on glucose. Using physiological properties of the parent wild type as constraints, elementary flux mode analysis of a large-scale model of the metabolism

  4. Enhancing sesquiterpene production in Saccharomyces cerevisiae through in silico driven metabolic engineering

    Asadollahi, Mohammadali; Maury, Jerome; Patil, Kiran Raosaheb

    2009-01-01

    A genome-scale metabolic model was used to identify new target genes for enhanced biosynthesis of sesquiterpenes in the yeast Saccharomyces cerevisiae. The effect of gene deletions on the flux distributions in the metabolic model of S. cerevisiae was assessed using OptGene as the modeling framework...

  5. Convexity Conditions and the Legendre-Fenchel Transform for the Product of Finitely Many Positive Definite Quadratic Forms

    Zhao Yunbin

    2010-01-01

    While the product of finitely many convex functions has been investigated in the field of global optimization, some fundamental issues such as the convexity condition and the Legendre-Fenchel transform for the product function remain unresolved. Focusing on quadratic forms, this paper is aimed at addressing the question: When is the product of finitely many positive definite quadratic forms convex, and what is the Legendre-Fenchel transform for it? First, we show that the convexity of the product is determined intrinsically by the condition number of so-called 'scaled matrices' associated with quadratic forms involved. The main result claims that if the condition number of these scaled matrices are bounded above by an explicit constant (which depends only on the number of quadratic forms involved), then the product function is convex. Second, we prove that the Legendre-Fenchel transform for the product of positive definite quadratic forms can be expressed, and the computation of the transform amounts to finding the solution to a system of equations (or equally, finding a Brouwer's fixed point of a mapping) with a special structure. Thus, a broader question than the open 'Question 11' in Hiriart-Urruty (SIAM Rev. 49, 225-273, 2007) is addressed in this paper.

  6. Interpreting expression data with metabolic flux models: predicting Mycobacterium tuberculosis mycolic acid production.

    Caroline Colijn

    2009-08-01

    Full Text Available Metabolism is central to cell physiology, and metabolic disturbances play a role in numerous disease states. Despite its importance, the ability to study metabolism at a global scale using genomic technologies is limited. In principle, complete genome sequences describe the range of metabolic reactions that are possible for an organism, but cannot quantitatively describe the behaviour of these reactions. We present a novel method for modeling metabolic states using whole cell measurements of gene expression. Our method, which we call E-Flux (as a combination of flux and expression, extends the technique of Flux Balance Analysis by modeling maximum flux constraints as a function of measured gene expression. In contrast to previous methods for metabolically interpreting gene expression data, E-Flux utilizes a model of the underlying metabolic network to directly predict changes in metabolic flux capacity. We applied E-Flux to Mycobacterium tuberculosis, the bacterium that causes tuberculosis (TB. Key components of mycobacterial cell walls are mycolic acids which are targets for several first-line TB drugs. We used E-Flux to predict the impact of 75 different drugs, drug combinations, and nutrient conditions on mycolic acid biosynthesis capacity in M. tuberculosis, using a public compendium of over 400 expression arrays. We tested our method using a model of mycolic acid biosynthesis as well as on a genome-scale model of M. tuberculosis metabolism. Our method correctly predicts seven of the eight known fatty acid inhibitors in this compendium and makes accurate predictions regarding the specificity of these compounds for fatty acid biosynthesis. Our method also predicts a number of additional potential modulators of TB mycolic acid biosynthesis. E-Flux thus provides a promising new approach for algorithmically predicting metabolic state from gene expression data.

  7. Metabolic activity, urease production, antibiotic resistance and virulence in dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus

    Vandecandelaere, Ilse; Van Nieuwerburgh, Filip; Deforce, Dieter

    2017-01-01

    In this paper, the metabolic activity in single and dual species biofilms of Staphylococcus epidermidis and Staphylococcus aureus isolates was investigated. Our results demonstrated that there was less metabolic activity in dual species biofilms compared to S. aureus biofilms. However, this was not observed if S. aureus and S. epidermidis were obtained from the same sample. The largest effect on metabolic activity was observed in biofilms of S. aureus Mu50 and S. epidermidis ET-024. A transcriptomic analysis of these dual species biofilms showed that urease genes and genes encoding proteins involved in metabolism were downregulated in comparison to monospecies biofilms. These results were subsequently confirmed by phenotypic assays. As metabolic activity is related to acid production, the pH in dual species biofilms was slightly higher compared to S. aureus Mu50 biofilms. Our results showed that S. epidermidis ET-024 in dual species biofilms inhibits metabolic activity of S. aureus Mu50, leading to less acid production. As a consequence, less urease activity is required to compensate for low pH. Importantly, this effect was biofilm-specific. Also S. aureus Mu50 genes encoding virulence-associated proteins (Spa, SplF and Dps) were upregulated in dual species biofilms compared to monospecies biofilms and using Caenorhabditis elegans infection assays, we demonstrated that more nematodes survived when co-infected with S. epidermidis ET-024 and S. aureus mutants lacking functional spa, splF or dps genes, compared to nematodes infected with S. epidermidis ET-024 and wild- type S. aureus. Finally, S. epidermidis ET-024 genes encoding resistance to oxacillin, erythromycin and tobramycin were upregulated in dual species biofilms and increased resistance was subsequently confirmed. Our data indicate that both species in dual species biofilms of S. epidermidis and S. aureus influence each other’s behavior, but additional studies are required necessary to elucidate the exact

  8. Reconstruction of a metabolic regulatory network in Escherichia coli for purposeful switching from cell growth mode to production mode in direct GABA fermentation from glucose.

    Soma, Yuki; Fujiwara, Yuri; Nakagawa, Takuya; Tsuruno, Keigo; Hanai, Taizo

    2017-09-01

    γ-aminobutyric acid (GABA) is a drug and functional food additive and is used as a monomer for producing the biodegradable plastic, polyamide 4. Recently, direct GABA fermentation from glucose has been developed as an alternative to glutamate-based whole cell bioconversion. Although total productivity in fermentation is determined by the specific productivity and cell amount responsible for GABA production, the optimal metabolic state for GABA production conflicts with that for bacterial cell growth. Herein, we demonstrated metabolic state switching from the cell growth mode based on the metabolic pathways of the wild type strain to a GABA production mode based on a synthetic metabolic pathway in Escherichia coli through rewriting of the metabolic regulatory network and pathway engineering. The GABA production mode was achieved by multiple strategies such as conditional interruption of the TCA and glyoxylate cycles, engineering of GABA production pathway including a bypass for precursor metabolite supply, and upregulation of GABA transporter. As a result, we achieved 3-fold improvement in total GABA production titer and yield (4.8g/L, 49.2% (mol/mol glucose)) in batch fermentation compared to the case without metabolic state switching (1.6g/L, 16.4% (mol/mol glucose)). This study reports the highest GABA production performance among previous reports on GABA fermentation from glucose using engineered E. coli. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  9. Metabolic engineering of Escherichia coli for ethanol production without foreign genes

    Kim, Youngnyun

    Worldwide dependence on finite petroleum-based energy necessitates alternative energy sources that can be produced from renewable resources. A successful example of an alternative transportation fuel is bioethanol, produced by microorganisms, from corn starch that is blended with gasoline. However, corn, currently the main feedstock for bioethanol production, also occupies a significant role in human food and animal feed chains. As more corn is diverted to bioethanol, the cost of corn is expected to increase with an increase in the price of food, feed and ethanol. Using lignocellulosic biomass for ethanol production is considered to resolve this problem. However, this requires a microbial biocatalyst that can ferment hexoses and pentoses to ethanol. Escherichia coli is an efficient biocatalyst that can use all the monomeric sugars in lignocellulose, and recombinant derivatives of E. coli have been engineered to produce ethanol as the major fermentation product. In my study, ethanologenic E. coli strains were isolated from a ldhA-, pflB- derivative without introduction of foreign genes. These isolates grew anaerobically and produced ethanol as the main fermentation product. The mutation responsible for anaerobic growth and ethanol production was mapped in the lpdA gene and the mutation was identified as E354K in three of the isolates tested. Another three isolates carried an lpdA mutation, H352Y. Enzyme kinetic studies revealed that the mutated form of the dihydrolipoamide dehydrogenase (LPD) encoded by the lpdA was significantly less sensitive to NADH inhibition than the native LPD. This reduced NADH sensitivity of the mutated LPD was translated into lower sensitivity to NADH of the pyruvate dehydrogenase complex in strain SE2378. The net yield of 4 moles of NADH and 2 moles of acetyl-CoA per mole of glucose produced by a combination of glycolysis and PDH provided a logical basis to explain the production of 2 moles of ethanol per glucose. The development of E

  10. Optimization of 64Cu production for Radio-metabolic therapy and for PET

    Groppi, F.; Bonardi, M.; Birattari, C.; Gini, L.; Guariglia, S.; Mainardi, C.; Menapace, E.

    2002-01-01

    The 6 4C u is a very interesting radioisotope for nuclear biomedical applications, due to its physical characteristics: a) it decay β+ and β- with maximum energy equal to 653 keV and 578 keV respectively, that correspond to maximum range of β+ in soft tissue ∼ 2.7 mm and average range ∼ 1 mm. In this way it is possible to give a local dose in the region of interest and to have a high resolution in the PET images; b) it has an half life of 12.7 h, that is suitable for medical applications, reducing the exposure of personnel and of the patients and the problem of waste discharge; c) besides the gamma of annihilation (36%), it has only one γ emission at 1345.84 keV (0.473%) with low intensity: this reduces the dose to healthy tissue and to medical personnel. For these reasons 6 4C u can be considered a dual purpose radionuclide: it can be used both to localise and measure the tumour mass through the PET tomography and for the metabolic therapy of the tumour itself. Cu(2.) forms a class of stable classical complexes and chelates with ditio carbamates (DTC), like ethylmethyl-DTC and tiosemicarbazonates, like pyruvaldehyde-bis-(N 4 -methyl semicarbazone (PTSM) and acetyl-bis-(N 4 -methyl semicarbazone) (ATSM), and finally with aza macrocyclic chelants (cyclens, cyclams, sarcofands), like DOTA, DOTP and SarAr). The behaviour of some of them has been already investigated in both cell cultures, rats and humans since the end of the 90-ties

  11. Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica

    Komi Nambou

    2015-12-01

    Full Text Available Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species.

  12. Cesium release from ceramic waste form materials in simulated canister corrosion product containing solutions

    Vittorio, Luca; Drabarek, Elizabeth; Chronis, Harriet; Griffith, Christopher S

    2004-07-01

    It has previously been demonstrated that immobilization of Cs{sup +} and/or Sr{sup 2+} sorbed on hexagonal tungsten oxide bronze (HTB) adsorbent materials can be achieved by heating the materials in air at temperatures in the range 500 - 1300 deg C. Highly crystalline powdered HTB materials formed by heating at 800 deg C show leach characteristics comparable to Cs-containing hot-pressed hollandites in the pH range from 0 to 12. As a very harsh leaching test, and also to model in a basic manner, leaching in the presence of canister corrosion products in oxidising environments, leaching of the bronzoid phases has been undertaken in Fe(NO{sub 3}){sub 3} solutions of increasing concentration. This is done in comparison with Cs -hollandite materials in order to compare the leaching characteristics of these two materials under such conditions. Both the Cs-loaded bronze and hollandite materials leach severely in Fe(NO{sub 3}){sub 3} losing virtually all of the immobilized Cs in a period of four days at 150 deg C. Total release of Cs and conversion of hollandite to titanium and iron titanium oxides begins to be observed at relatively low concentrations and is virtually complete after four days reaction in 0.5 mol/L Fe(NO{sub 3}){sub 3}. In the case of the bronze, all of the Cs is also extracted but the HTB structure is preserved. The reaction presumably involves an ion-exchange mechanism and iron oxide with a spinel structure is also observed at high Fe concentrations. (authors)

  13. Cesium release from ceramic waste form materials in simulated canister corrosion product containing solutions

    Vittorio, Luca; Drabarek, Elizabeth; Chronis, Harriet; Griffith, Christopher S.

    2004-01-01

    It has previously been demonstrated that immobilization of Cs + and/or Sr 2+ sorbed on hexagonal tungsten oxide bronze (HTB) adsorbent materials can be achieved by heating the materials in air at temperatures in the range 500 - 1300 deg C. Highly crystalline powdered HTB materials formed by heating at 800 deg C show leach characteristics comparable to Cs-containing hot-pressed hollandites in the pH range from 0 to 12. As a very harsh leaching test, and also to model in a basic manner, leaching in the presence of canister corrosion products in oxidising environments, leaching of the bronzoid phases has been undertaken in Fe(NO 3 ) 3 solutions of increasing concentration. This is done in comparison with Cs -hollandite materials in order to compare the leaching characteristics of these two materials under such conditions. Both the Cs-loaded bronze and hollandite materials leach severely in Fe(NO 3 ) 3 losing virtually all of the immobilized Cs in a period of four days at 150 deg C. Total release of Cs and conversion of hollandite to titanium and iron titanium oxides begins to be observed at relatively low concentrations and is virtually complete after four days reaction in 0.5 mol/L Fe(NO 3 ) 3 . In the case of the bronze, all of the Cs is also extracted but the HTB structure is preserved. The reaction presumably involves an ion-exchange mechanism and iron oxide with a spinel structure is also observed at high Fe concentrations. (authors)

  14. Towards a sustainable bio-based economy: Redirecting primary metabolism to new products with plant synthetic biology.

    Shih, Patrick M

    2018-08-01

    Humans have domesticated many plant species as indispensable sources of food, materials, and medicines. The dawning era of synthetic biology represents a means to further refine, redesign, and engineer crops to meet various societal and industrial needs. Current and future endeavors will utilize plants as the foundation of a bio-based economy through the photosynthetic production of carbohydrate feedstocks for the microbial fermentation of biofuels and bioproducts, with the end goal of decreasing our dependence on petrochemicals. As our technological capabilities improve, metabolic engineering efforts may expand the utility of plants beyond sugar feedstocks through the direct production of target compounds, including pharmaceuticals, renewable fuels, and commodity chemicals. However, relatively little work has been done to fully realize the potential in redirecting central carbon metabolism in plants for the engineering of novel bioproducts. Although our ability to rationally engineer and manipulate plant metabolism is in its infancy, I highlight some of the opportunities and challenges in applying synthetic biology towards engineering plant primary metabolism. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Effects of weekly regrouping of prepartum dairy cows on metabolic, health, reproductive, and productive parameters.

    Silva, P R B; Moraes, J G N; Mendonça, L G D; Scanavez, A A; Nakagawa, G; Fetrow, J; Endres, M I; Chebel, R C

    2013-07-01

    The objectives of the current experiment were to determine the effect of 2 prepartum grouping strategies on the health, metabolic, reproductive, and productive parameters of dairy cows. Jersey cows enrolled in the experiment at 253±3 d of gestation (d 0=calving) were balanced for parity and projected 305-d mature equivalent and assigned to 1 of 2 treatments. Cows assigned to the traditional (TRD; n=6 replicates with a total of 308 cows) treatment were moved to the study pen as a group of 44 cows and weekly thereafter groups of 2 to 15 cows were moved to the study pen to reestablish stocking density. Cows assigned to the all-in-all-out (AIAO; n=6 replicates with a total of 259 cows) treatment were moved to the study pen in groups of 44 cows, but no new cows entered the AIAO pen until the end of the replicate. At the end of each replicate, a new TRD and AIAO group started but pens were switched. Cows were milked thrice daily and monthly milk yield, fat and protein contents, and somatic cell count data were recorded up to 305 d postpartum. Plasma nonesterified fatty acid concentration was measured weekly from d -18±3 to 24±3 and plasma β-hydroxybutyrate was measured weekly from d 3±3 to 24±3. Cows were examined on d 1, 4±1, 7±1, 10±1, and 13±1 for diagnosis of uterine diseases and had their ovaries scanned by ultrasound on d 39±3 and 53±3 to determine resumption of ovarian cycles. Average stocking density was reduced for the AIAO (71.9%) treatment compared with the TRD (86.9%) treatment. Treatment did not affect the incidences of retained fetal membranes (TRD=10.9, AIAO=11.6%), metritis (TRD=16.7, AIAO=19.8%), and acute metritis (TRD=1.7, AIAO=3.6%). Concentrations of nonesterified fatty acids (TRD=80.4±8.2, AIAO=62.9±8.5 µmol/L) and β-hydroxybutyrate (TRD=454.4±10.9, AIAO=446.1±11.1 µmol/L) were not different between treatments. Percentages of cows that resumed ovarian cycles by d 39±3 (TRD=70.8, AIAO=63.1%) and 53±3 (TRD=90.1, AIAO=90.2%) were

  16. PSEUDOAFFINITY CHROMATOGRAPHY ENRICHMENT OF GLYCATED PEPTIDES FOR MONITORING ADVANCED GLYCATION END PRODUCTS (AGES IN METABOLIC DISORDERS

    Rajasekar R. Prasanna

    2016-09-01

    Full Text Available Advanced Glycation End (AGE products are produced due to diabetic progression and they are responsible for many complications in the diabetic disorder. The diabetic progression is measured, particularly following glycated hemoglobin using specific antibodies. However, the most abundant protein in blood, human serum albumin, is also found to be glycated which has a much shorter half life and gives information on short term glycemic control. In addition, glycated albumins are considered as markers of diabetic complications such as nephropathy, peripheral vascular calcification and also in Alzheimer’s disease. The glycation proceeds from the interaction between aldehyde group of sugar and the free amino group of the protein, resulting in the formation of Schiff’s base, which undergoes a series of modifications leading to generation of imidazoyl derivatives of amino acids known as Amadori rearrangement products. The imidazoyl derivatives from arginine and lysine are the most prominent modifications observed in proteins in the presence of reducing sugar and these imidazoyl derivatives have an affinity towards certain transition metal ions. Based on our earlier exhaustive work on trapping the histidine peptides using transition metal ion, Cu(II linked to imino-diacetate complex, we explored Cu(II immobilized metal affinity chromatography (IMAC as a potential tool for specific detection of glycated peptides of human serum albumin. Our results clearly demonstrate that Cu(II IMAC is able to detect glycated peptides very efficiently while the non-glycated forms were not retained on the Cu (II column as confirmed by LC-MS/MS analysis. We further discuss the utility of IMAC technology to enrich the detection of AGE products in plasma. We anticipate that these studies may provide valuable information on understanding disease pathologies and the potential of AGE products as biomarkers of various diseases including neurodegenerative, renal and

  17. Adiposity indicators lipid accumulation product and triglyceride-glucose index as alternate criteria for the diagnosis of metabolic obesity in adult

    Mariya Tabassum; Md. Matiur Rahman; Miliva Mozaffor

    2018-01-01

    Metabolic obesity refers to the state of having metabolic syndrome irrespective of one’s BMI. This study was aimed to elucidate the lipid accumulation product and triglyceride-glucose index as simple and alternate criteria for the detecting metabolic obesity in adult. The study was conducted in 200 adult (age range: 19-45 years). According to lipid accumulation product and Triglyceride-glucose index, the prevalence of metabolic obesity was 54.0% and 53.5% respectively. With a cutoff value of ...

  18. Effects of different starch sources on metabolic profile, production and fertility parameters in dairy cows.

    Mikuła, R; Nowak, W; Jaśkowski, J M; Maćkowiak, P; Oszmałek, E Pruszyńska

    2011-01-01

    The objective of the study was to determine the effect of replacing triticale (high rumen degradable starch) with maize grain (low rumen degradable starch) during the transition period and the first 120 days of lactation on metabolic and hormonal profile indices, milk production and fertility performance in cows. Forty-eight Holstein-Friesian dairy cows were divided into 4 groups: TT (2.5 kg triticale grain/cow per day supplemented from 14 days prepartum to day 120 postpartum), TM (2.5 kg triticale grain/cow per day supplemented from day 14 before parturition to calving, and then 2.5 kg maize grain to 120 days of lactation), MT (2.5 kg maize grain/cow per day supplemented from day 14 before parturition to calving, and then 2.5 kg triticale grain to 120 days of lactation), MM (2.5 kg maize grain/cow per day supplemented from 14 days prepartum to day 120 postpartum). Blood samples were collected 3 weeks and 1 week before calving and on days 14, 56 and 70 of lactation, and they were analyzed in terms of concentrations of glucose, insulin, leptin, insulin-like growth factor I, nonesterified fatty acids, triglycerides, cholesterol, blood urea nitrogen and activities of aspartate aminotransferase and gamma glutamyl transpeptidase. Milk samples were collected twice a day at weekly intervals and analyzed for fat, protein and lactose. Milk yield and individual dry mater intake were recorded at weekly intervals. Body condition was estimated 3 weeks before calving, on parturition day and on days 14, 56 and 120 of lactation. Replacing triticale grain with maize grain in the transition period and during lactation positively affected fertility of lactating cows. An increased first service conception rate and shortening of the days open period was observed in MM and TM groups in comparison to those found in group MT (P cows than triticale grain.

  19. Die design optimization on sheet metal forming with considering the phenomenon of springback to improve product quality

    Darmawan Agung Setyo

    2018-01-01

    Full Text Available The process of sheet metal forming is one of the very important processes in manufacture of products mainly in the automotive field. In sheet metal forming, it is added a certain size at the die to tolerate a result of the elasticity restoration of material. Therefore, when the product is removed from the die then the process elastic recovery will end within the allowable tolerance size. Extra size of the die is one method to compensate for springback. The aim of this research is to optimize the die by entering a springback value in die design to improve product quality that is associated with accuracy the final size of the product. Simulation processes using AutoForm software are conducted to determine the optimal parameters to be used in the forming process. Variations the Blank Holder Force of 77 N, 97 N, and 117 N are applied to the plate material. The Blank Holder Force application higher than 97 N cannot be conducted because the Forming Limit Diagram indicates the risk of tearing. Then the Blank Holder Force of 37 N, 57 N and 77 N are selected and applied in cup drawing process. Even though a few of wrinkling are appear, however there is no significant deviation of dimension between the product and the design of cup.

  20. Production of (+)-valencene in the mushroom-forming fungus S. commune

    Scholtmeijer, K.; Beekwilder, J.; Cankar, K.; Wösten, H.A.B.; Lugones, L.G.; Bosch, H.J.

    2014-01-01

    Production of commercially interesting sesquiterpenes was previously examined in plants and microorganisms such as Escherichia coli and Saccharomyces cerevisiae. We here investigate the potential of the mushroom Schizophyllum commune for the production of sesquiterpenes. Genomic analysis of S.

  1. The production of (14C) oxalate during the metabolism of (14C) carbohydrates in isolated rat hepatocytes.

    Rofe, A M; James, H M; Bais, R; Edwards, J B; Conyers, R A

    1980-04-01

    Oxalate (14C) was produced during the metabolism of (U-14C) carbohydrates in hepatocytes isolated from normal rats. At 10 mM, the order of oxalate production was fructose > glycerol > xylitol > sorbitol greater than or equal to glucose in the ratio 10 : 4 : 3 : 1 : 1. This difference between oxalate production from fructose and glucose was reflected in their rates of utilisation, glucose being poorly metabolised in hepatocytes from fasted rats. Fructose was rapidly metabolised, producing glucose, lactate and pyruvate as the major metabolites. Glycerol, xylitol and sorbitol were metabolised at half the rate of fructose, the major metabolites being glucose, lactate and glycerophosphate. The marked similarity in the pattern of intermediary metabolites produced by these polyols was not, however, reflected in the rates of oxalate production. Hepatic polyol metabolism resulted in high levels of cytosolic NADH, as indicated by elevated lactate : pyruvate and glycerophosphate : dihydroxyacetone phosphate ratios. The artificial electron acceptor, phenazine methosulphate (PMS) stimulated oxalate production from the polyols, particularly xylitol. In the presence of PMS, the order of oxalate production was fructose greater than or equal to xylitol > glycerol > sorbitol in the ratio 10 : 10 : 6 : 2. The production of glucose, lactate and pyruvate from the polyols was also stimulated by PMS, whereas the general metabolism of fructose, including oxalate production, was little affected. Oxalate (14C) was produced from (1-14C), (2-14C) and (6-14C) but not (3,4-14C) glucose in hepatocytes isolated from non-fasted, pyridoxine-deficient rats. Whilst this labelling pattern is consistent with oxalate being produced by a number of pathways, it is suggested that metabolism via hydroxypyruvate is a major route for oxalate production from various carbohydrates, with perhaps the exception of xylitol, which appears to have an alternative mechanism for oxalate production. The observation that

  2. Green Consumerism : an Eco-Friendly Behaviour Form Through The Green Product Consumption and Green Marketing

    Handayani, Wiwik

    2017-01-01

    This research is referred to analyse the influence of consumer attitude of green product towards purchase intention. The consumer attitude of green product is a psychological tendencies that is expressed by evaluating a certain entity with some advantage or disadvantage considerations. The problem of this research is the low of cunsumer awareness to consume green product, because the lack to comprehend the importance of green product usage for health and eco-friendly. The purpose of this rese...

  3. Advance Planning of Form Properties in the Written Production of Single and Multiple Words

    Damian, Markus F.; Stadthagen-Gonzalez, Hans

    2009-01-01

    Three experiments investigated the scope of advance planning in written production. Experiment 1 manipulated phonological factors in single word written production, and Experiments 2 and 3 did the same in the production of adjective-noun utterances. In all three experiments, effects on latencies were found which mirrored those previously…

  4. In silico method for modelling metabolism and gene product expression at genome scale

    Lerman, Joshua A.; Hyduke, Daniel R.; Latif, Haythem; Portnoy, Vasiliy A.; Lewis, Nathan E.; Orth, Jeffrey D.; Rutledge, Alexandra C.; Smith, Richard D.; Adkins, Joshua N.; Zengler, Karsten; Palsson, Bernard O.

    2012-07-03

    Transcription and translation use raw materials and energy generated metabolically to create the macromolecular machinery responsible for all cellular functions, including metabolism. A biochemically accurate model of molecular biology and metabolism will facilitate comprehensive and quantitative computations of an organism's molecular constitution as a function of genetic and environmental parameters. Here we formulate a model of metabolism and macromolecular expression. Prototyping it using the simple microorganism Thermotoga maritima, we show our model accurately simulates variations in cellular composition and gene expression. Moreover, through in silico comparative transcriptomics, the model allows the discovery of new regulons and improving the genome and transcription unit annotations. Our method presents a framework for investigating molecular biology and cellular physiology in silico and may allow quantitative interpretation of multi-omics data sets in the context of an integrated biochemical description of an organism.

  5. Correlation Between Metabolic Syndrome, Periodontitis and Reactive Oxygen Species Production. A Pilot Study

    Patini, Romeo; Gallenzi, Patrizia; Spagnuolo, Gianrico; Cordaro, Massimo; Cantiani, Monica; Amalfitano, Adriana; Arcovito, Alessandro; Callà, Cinzia; Mingrone, Gertrude; Nocca, Giuseppina

    2017-01-01

    Background and Objective: Metabolic syndrome (MetS) is associated with an increased risk of periodontitis even if the mechanism is unknown. Since both MetS and periodontitis are characterized by an alteration of inflammation status, the aim of this pilot study was to determine if differences in ROS metabolism of phagocytes isolated from (A) patients with MetS, (B) patients with both MetS and mild periodontitis, (C) healthy subjects and (D) normal weight subjects with mild periodontitis, were ...

  6. Recent advances in engineering propionyl-CoA metabolism for microbial production of value-added chemicals and biofuels.

    Srirangan, Kajan; Bruder, Mark; Akawi, Lamees; Miscevic, Dragan; Kilpatrick, Shane; Moo-Young, Murray; Chou, C Perry

    2017-09-01

    Diminishing fossil fuel reserves and mounting environmental concerns associated with petrochemical manufacturing practices have generated significant interests in developing whole-cell biocatalytic systems for the production of value-added chemicals and biofuels. Although acetyl-CoA is a common natural biogenic precursor for the biosynthesis of numerous metabolites, propionyl-CoA is unpopular and non-native to most organisms. Nevertheless, with its C3-acyl moiety as a discrete building block, propionyl-CoA can serve as another key biogenic precursor to several biological products of industrial importance. As a result, engineering propionyl-CoA metabolism, particularly in genetically tractable hosts with the use of inexpensive feedstocks, has paved an avenue for novel biomanufacturing. Herein, we present a systematic review on manipulation of propionyl-CoA metabolism as well as relevant genetic and metabolic engineering strategies for microbial production of value-added chemicals and biofuels, including odd-chain alcohols and organic acids, bio(co)polymers and polyketides. [Formula: see text].

  7. [Advances in metabolic engineering for the microbial production of naturally occurring terpenes-limonene and bisabolene: a mini review].

    Pang, Yaru; Hu, Zhihui; Xiao, Dongguang; Yu, Aiqun

    2018-01-25

    Limonene (C₁₀H₁₆) and bisabolene (C₁₅H₂₄) are both naturally occurring terpenes in plants. Depending on the number of C₅ units, limonene and bisabolene are recognized as representative monoterpenes and sesquiterpenes, respectively. Limonene and bisabolene are important pharmaceutical and nutraceutical products used in the prevention and treatment of cancer and many other diseases. In addition, they can be used as starting materials to produce a range of commercially valuable products, such as pharmaceuticals, nutraceuticals, cosmetics, and biofuels. The low abundance or yield of limonene and bisabolene in plants renders their isolation from plant sources non-economically viable. Isolation of limonene and bisabolene from plants also suffers from low efficiency and often requires harsh reaction conditions, prolonged reaction times, and expensive equipment cost. Recently, the rapid developments in metabolic engineering of microbes provide a promising alternative route for producing these plant natural products. Therefore, producing limonene and bisabolene by engineering microbial cells into microbial factories is becoming an attractive alternative approach that can overcome the bottlenecks, making it more sustainable, environmentally friendly and economically competitive. Here, we reviewed the status of metabolic engineering of microbes that produce limonene and bisabolene including microbial hosts, key enzymes, metabolic pathways and engineering of limonene/bisabolene biosynthesis. Furthermore, key challenges and future perspectives were discussed.

  8. Forming of the dynamics of the changes in convergent production system depending on size of production party

    Bożena Zwolińska

    2017-09-01

    Full Text Available Background: In terms of Lean Six Sigma, the whole process focuses on clients and their needs. Existence of a client generates the supply of companies. Extended customization has a negative impact for a structure of the production system. Dynamics of changes and no predictability of system’s state in time t+1 lead to increase of the operational costs. It particularly affects those companies which are producing goods using MTO (make – to – order method in short series. The goal of this article is to establish a mathematical model defining how the structure of a production system is subject to change depending on the volume of the production batch for a production system in accordance with MTO. Furthermore pilot calculations have been presented which determine the probability value, how subsequent random variables are contained within three standard deviations (±3δ from the determined expected value (ET for the entire production structure. Months of analysis and research on introducing selected lean toolbox components to a polish company from the small and medium enterprises sector resulted in the models presented in the article. The production structure of the discussed actual facility is complex and is of converged nature in accordance with MTO, while the final products are manufactured in short production series with a relatively wide customization options. Materials and results: Wrought models consider theories of  Klir and Maserovicz [Mesarovic 1964] and also theory of mass operation (one of the probability areas.  In the article there are results from two models which are fundamental in defining problems in logistics engineering and production in scientific research. Important attribute of presented models is a fact that they consider relations between variables in a structure of consecutive processes and also consider relations between a size of production party and a real object. Presented models are not only theoretical coverage

  9. Straight-chain halocarbon forming fluids for TRISO fuel kernel production – Tests with yttria-stabilized zirconia microspheres

    Baker, M.P. [Nuclear Science and Engineering Program, Metallurgical and Materials Engineering Department, Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); King, J.C., E-mail: kingjc@mines.edu [Nuclear Science and Engineering Program, Metallurgical and Materials Engineering Department, Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); Gorman, B.P. [Metallurgical and Materials Engineering Department, Colorado Center for Advanced Ceramics, Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States); Braley, J.C. [Nuclear Science and Engineering Program, Chemistry and Geochemistry Department, Colorado School of Mines, 1500 Illinois St., Golden, CO 80401 (United States)

    2015-03-15

    Highlights: • YSZ TRISO kernels formed in three alternative, non-hazardous forming fluids. • Kernels characterized for size, shape, pore/grain size, density, and composition. • Bromotetradecane is suitable for further investigation with uranium-based precursor. - Abstract: Current methods of TRISO fuel kernel production in the United States use a sol–gel process with trichloroethylene (TCE) as the forming fluid. After contact with radioactive materials, the spent TCE becomes a mixed hazardous waste, and high costs are associated with its recycling or disposal. Reducing or eliminating this mixed waste stream would not only benefit the environment, but would also enhance the economics of kernel production. Previous research yielded three candidates for testing as alternatives to TCE: 1-bromotetradecane, 1-chlorooctadecane, and 1-iodododecane. This study considers the production of yttria-stabilized zirconia (YSZ) kernels in silicone oil and the three chosen alternative formation fluids, with subsequent characterization of the produced kernels and used forming fluid. Kernels formed in silicone oil and bromotetradecane were comparable to those produced by previous kernel production efforts, while those produced in chlorooctadecane and iodododecane experienced gelation issues leading to poor kernel formation and geometry.

  10. Green Consumerism : an Eco-Friendly Behaviour Form Through The Green Product Consumption and Green Marketing

    Wiwik Handayani

    2017-09-01

    Full Text Available This research is referred to analyse the influence of consumer attitude of green product towards purchase intention. The consumer attitude of green product is a psychological tendencies that is expressed by evaluating a certain entity with some advantage or disadvantage considerations. The problem of this research is the low of cunsumer awareness to consume green product, because the lack to comprehend the importance of green product usage for health and eco-friendly. The purpose of this research is to test the influence of consumer attitude of green products towards purchase intention. Hypothesis testing using Partial Least Square (PLS. The result of analysis show that there is influence among consumer attitude of green product towards consumer purchase intention significantly.

  11. Aldosterone synthase gene polymorphism in alimentary obesity, metabolic syndrome components, some secondary forms of arterial hypertension, pathology of the adrenals glands core (literature review

    S.N. Koval

    2017-08-01

    Full Text Available Hormonal factors of adrenal origin belong to the pathophysiological mechanisms of the formation and progression of arterial hypertension (AH and should be consi­dered while developing differentiated approaches to the treatment and prevention of hypertensive states, their primary, secondary and resistant forms. The first thing we should point up is aldosterone (AL, enzyme aldosterone synthase (AS, which takes a direct part in the formation of this hormone, as well as gene polymorphisms of AS, which have not only molecular genetic, but also differential diagnostic and therapeutic significance for secondary forms of arterial hypertension, abdominal obesity (AO, metabolic syndrome (MS, adrenal pathology and other endocrine disorders. AL is a steroid (mineralocorticoid hormone of the adrenal cortex, which is synthesized from cholesterol (CH, mainly in the glomerular zone of the adrenal glands, is released under the action of angiotensin II (A II and potassium ions (K+. AL acti­vity is mediated through the corresponding mineralocorticoid receptors (MKR. The particular importance in AH and MS development belongs to AL activation and MKR density in adipocytes, this phenomenon is accompanied by increased expression of pro-inflammatory cytokines, leptin, an adipogenic effect, and the inhibition of MCR activity is accompanied by increased production of adiponectin, which is more pronounced in patients with AH. Aldosterone synthase, a mitochondrial human enzyme encoded by the CYP11B2 gene (cytochrome P450, family 11, subfamily B, polypeptide 2 is located on the 8th chromosome. AS belongs to the superfamily of cytochrome P450 and regulates the synthesis of AL hormone. The CYP11B2 gene encodes the key enzyme for the synthesis of AL 18-hydroxylase. In scientific papers, single nucleotide polymorphism (SNP of AS gene is often studied, such as 5312T, Intron 2, Lys-173/Arg; T-344C, 3097 C/A. 227 SNP of the AS gene were identified in different

  12. Insight into the product film formed on Ni-advanced weathering steel in a tropical marine atmosphere

    Wu, Wei; Cheng, Xuequn; Hou, Huaxing; Liu, Bo; Li, Xiaogang

    2018-04-01

    The product film formed on Ni-advanced weathering steel in a tropical marine environment was investigated in detail through outdoor exposure by using diverse surface analysis techniques combined with electrochemical impedance spectroscopy and scanning kelvin probe measurements. The results showed that the product film was mainly composed of nanophasic goethite in the inner layer and maghemite, akaganeite, and hematite in the outer layer. Moreover, the resistance to atmospheric corrosion gradually increased from the outermost product film to the innermost film. Ni was significantly enriched in the inner layer in the form of the spinel phase NiFe2O4, which transformed lepidocrocite to fine-grained goethite, withstood the invasion of chloridion, and improved the corrosion potential of the product film in a tropical marine atmosphere.

  13. Friction and lubrication modelling in sheet metal forming: Influence of lubrication amount, tool roughness and sheet coating on product quality

    Hol, J.; Wiebenga, J. H.; Carleer, B.

    2017-09-01

    In the stamping of automotive parts, friction and lubrication play a key role in achieving high quality products. In the development process of new automotive parts, it is therefore crucial to accurately account for these effects in sheet metal forming simulations. This paper presents a selection of results considering friction and lubrication modelling in sheet metal forming simulations of a front fender product. For varying lubrication conditions, the front fender can either show wrinkling or fractures. The front fender is modelled using different lubrication amounts, tool roughness’s and sheet coatings to show the strong influence of friction on both part quality and the overall production stability. For this purpose, the TriboForm software is used in combination with the AutoForm software. The results demonstrate that the TriboForm software enables the simulation of friction behaviour for varying lubrication conditions, i.e. resulting in a generally applicable approach for friction characterization under industrial sheet metal forming process conditions.

  14. Advances in metabolic pathway and strain engineering paving the way for sustainable production of chemical building blocks

    Chen, Yun; Nielsen, Jens

    2013-01-01

    Bio-based production of chemical building blocks from renewable resources is an attractive alternative to petroleum-based platform chemicals. Metabolic pathway and strain engineering is the key element in constructing robust microbial chemical factories within the constraints of cost effective...... production. Here we discuss how the development of computational algorithms, novel modules and methods, omics-based techniques combined with modeling refinement are enabling reduction in development time and thus advance the field of industrial biotechnology. We further discuss how recent technological...

  15. Application of Specific Features of Industrial Products when Forming and Developing Brands of Industrial Enterprises

    Yatsentiuk Stanislav V.

    2014-03-01

    Full Text Available The article analyses and structures approaches and principles of formulation of industrial products. It offers classification of goods and markets of industrial products by their characteristics and participants. It identifies main participants that make decisions at B2C and B2B markets and characterises their specific features and motivation when making decisions on purchase of products of industrial enterprises. It studies and analyses indicators of development of domestic markets of consumer goods and market of industrial products and dynamics of development of their relation in retrospective view.

  16. The effect of feed rations containing high moisture crimped corn ensiled with microbial inoculant or chemical additive on milk production and metabolism of dairy cows

    László Könyves

    2015-01-01

    Full Text Available The study evaluated the effects of crimped corn preserved either with organic acids or with a microbial inoculant on a range of metabolic and production indicators of dairy cows. Two hundred and sixty in-calf, second and third parity cows were selected into pairs on basis of age, parity, milk production in previous lactation, days in milk and body condition score with the greatest possible conformity within pairs. Cow pairs were assigned into a 2-period crossover experiment (2 × 45 days and kept in separate groups within the same shed. Dietary treatments were TMR with crimped corn preserved with either organic acids (treatment K or microbial inoculant (treatment B. Ten superbly matched cow-pairs were selected to form nucleus pairs for metabolic studies. The preservatives had no effect on the nutrient content of crimped corn. Crimped corn preserved with the microbial inoculant were found mouldy, predominantly with Mucor sp. at a number high enough to inhibit the growth of lactic acid bacteria, and had significantly higher pH and ammonium concentration compared to the chemical treatment. The milk yield of treatment K cows was significantly higher than that of treatment B cows with identical feed intake. Blood beta-hydroxy-butyrate concentration was lower and blood aspartate amino transferase activity higher with treatment K compared to treatment B. Results of this study suggest the superiority of total mixed rations containing chemically preserved crimped corn in terms of ammonia and microbiological indicators of crimped corn, significantly higher milk yield, and balanced energy metabolism.

  17. Deletion of genes involved in glutamate metabolism to improve poly-gamma-glutamic acid production in B. amyloliquefaciens LL3.

    Zhang, Wei; He, Yulian; Gao, Weixia; Feng, Jun; Cao, Mingfeng; Yang, Chao; Song, Cunjiang; Wang, Shufang

    2015-02-01

    Here, we attempted to elevate poly-gamma-glutamic acid (γ-PGA) production by modifying genes involved in glutamate metabolism in Bacillus amyloliquefaciens LL3. Products of rocR, rocG and gudB facilitate the conversion from glutamate to 2-oxoglutarate in Bacillus subtillis. The gene odhA is responsible for the synthesis of a component of the 2-oxoglutarate dehydrogenase complex that catalyzes the oxidative decarboxylation of 2-oxoglutarate to succinyl coenzyme A. In-frame deletions of these four genes were performed. In shake flask experiments the gudB/rocG double mutant presented enhanced production of γ-PGA, a 38 % increase compared with wild type. When fermented in a 5-L fermenter with pH control, the γ-PGA yield of the rocR mutant was increased to 5.83 g/L from 4.55 g/L for shake flask experiments. The gudB/rocG double mutant produced 5.68 g/L γ-PGA compared with that of 4.03 g/L for the wild type, a 40 % increase. Those results indicated the possibility of improving γ-PGA production by modifying glutamate metabolism, and identified potential genetic targets to improve γ-PGA production.

  18. Effects of NADH-preferring xylose reductase expression on ethanol production from xylose in xylose-metabolizing recombinant Saccharomyces cerevisiae.

    Lee, Sung-Haeng; Kodaki, Tsutomu; Park, Yong-Cheol; Seo, Jin-Ho

    2012-04-30

    Efficient conversion of xylose to ethanol is an essential factor for commercialization of lignocellulosic ethanol. To minimize production of xylitol, a major by-product in xylose metabolism and concomitantly improve ethanol production, Saccharomyces cerevisiae D452-2 was engineered to overexpress NADH-preferable xylose reductase mutant (XR(MUT)) and NAD⁺-dependent xylitol dehydrogenase (XDH) from Pichia stipitis and endogenous xylulokinase (XK). In vitro enzyme assay confirmed the functional expression of XR(MUT), XDH and XK in recombinant S. cerevisiae strains. The change of wild type XR to XR(MUT) along with XK overexpression led to reduction of xylitol accumulation in microaerobic culture. More modulation of the xylose metabolism including overexpression of XR(MUT) and transaldolase, and disruption of the chromosomal ALD6 gene encoding aldehyde dehydrogenase (SX6(MUT)) improved the performance of ethanol production from xylose remarkably. Finally, oxygen-limited fermentation of S. cerevisiae SX6(MUT) resulted in 0.64 g l⁻¹ h⁻¹ xylose consumption rate, 0.25 g l⁻¹ h⁻¹ ethanol productivity and 39% ethanol yield based on the xylose consumed, which were 1.8, 4.2 and 2.2 times higher than the corresponding values of recombinant S. cerevisiae expressing XR(MUT), XDH and XK only. Copyright © 2011 Elsevier B.V. All rights reserved.

  19. Heavy flavours production in quark-gluon plasma formed in high energy nuclear reactions

    Kloskinski, J.

    1985-01-01

    Results on compression and temperatures of nuclear fireballs and on relative yield of strange and charmed hadrons are given . The results show that temperatures above 300 MeV and large compressions are unlikely achieved in average heavy ion collision. In consequence, thermal production of charm is low. Strange particle production is, however, substantial and indicates clear temperature - threshold behavior.

  20. Comparison of Lipid Accumulation Product Index with Body Mass Index and Waist Circumference as a Predictor of Metabolic Syndrome in Indian Population.

    Ray, Lopamudra; Ravichandran, Kandasamy; Nanda, Sunil Kumar

    2018-06-01

    Metabolic syndrome (MetS), which confers a high risk for cardiovascular diseases, needs early diagnosis and treatment to reduce morbidity and mortality. Lipid accumulation product index has been reported to be an inexpensive marker of visceral fat and metabolic syndrome. This study aimed to evaluate lipid accumulation product index as a marker for metabolic syndrome in the Indian population where the prevalence of the condition is steadily increasing. A hospital-based, case-control study was conducted with 72 diagnosed cases of metabolic syndrome and 79 control subjects. In all the participants, body mass index (BMI) and lipid accumulation product index were calculated. The difference between cases and controls in BMI, waist circumference (WC), and lipid accumulation product index was assessed by Mann-Whitney U test/unpaired t-test. Associations of BMI, WC, and lipid accumulation product index with metabolic syndrome were compared by multiple logistic regression analysis and receiver operating characteristic analysis. BMI, WC, and lipid accumulation product index were significantly higher in metabolic syndrome (P product index had the highest prediction accuracy. The parameter also had a high area under curve of 0.901 (95% confidence interval 0.85-0.95) and a high sensitivity (76.4%), specificity (91.1%), positive predictive value (88.7%), and negative predictive value (80.9%) for detection of metabolic syndrome. In the Indian population, lipid accumulation product index is a better predictor of metabolic syndrome compared to BMI and WC and should be incorporated in laboratory reports as early, accurate, and inexpensive indicator of metabolic syndrome.

  1. Recent advances in the metabolic engineering of Corynebacterium glutamicum for the production of lactate and succinate from renewable resources.

    Tsuge, Yota; Hasunuma, Tomohisa; Kondo, Akihiko

    2015-03-01

    Recent increasing attention to environmental issues and the shortage of oil resources have spurred political and industrial interest in the development of environmental friendly and cost-effective processes for the production of bio-based chemicals from renewable resources. Thus, microbial production of commercially important chemicals is viewed as a desirable way to replace current petrochemical production. Corynebacterium glutamicum, a Gram-positive soil bacterium, is one of the most important industrial microorganisms as a platform for the production of various amino acids. Recent research has explored the use of C. glutamicum as a potential cell factory for producing organic acids such as lactate and succinate, both of which are commercially important bulk chemicals. Here, we summarize current understanding in this field and recent metabolic engineering efforts to develop C. glutamicum strains that efficiently produce L- and D-lactate, and succinate from renewable resources.

  2. Metabolic characterization and transformation of the non-dairy Lactococcus lactis strain KF147, for production of ethanol from xylose

    Petersen, Kia Vest; Liu, Jianming; Chen, Jun

    2017-01-01

    producing ethanol as the sole fermentation product with a high yield corresponding to 83% of the theoretical maximum. The results clearly indicate the great potential of using the more metabolically diverse non-dairy L. lactis strains for bio-production based on xylose containing feedstocks.......The non-dairy lactic acid bacterium Lactococcus lactis KF147 can utilize xylose as the sole energy source. To assess whether KF147 could serve as a platform organism for converting second generation sugars into useful chemicals, we characterized growth and product formation for KF147 when grown...... the arcA gene encoding the arginine deiminase. The fermentation product profile suggested two routes for xylose degradation, the phosphoketolase pathway and the pentose phosphate pathway. Inactivation of the phosphoketolase pathway redirected the entire flux through the pentose phosphate pathway whereas...

  3. The Production of Advanced Glass Ceramic HLW Forms using Cold Crucible Induction Melter

    Veronica J Rutledge; Vince Maio

    2013-10-01

    Cold Crucible Induction Melters (CCIMs) will favorably change how High-Level radioactive Waste (from nuclear fuel recovery) is treated in the 21st century. Unlike the existing Joule-Heated Melters (JHMs) currently in operation for the glass-based immobilization of High-Level Waste (HLW), CCIMs offer unique material features that will increase melt temperatures, increase throughput, increase mixing, increase loading in the waste form, lower melter foot prints, eliminate melter corrosion and lower costs. These features not only enhance the technology for producing HLW forms, but also provide advantageous attributes to the waste form by allowing more durable alternatives to glass. This paper discusses advantageous features of the CCIM, with emphasis on features that overcome the historical issues with the JHMs presently utilized, as well as the benefits of glass ceramic waste forms over borosilicate glass waste forms. These advantages are then validated based on recent INL testing to demonstrate a first-of-a-kind formulation of a non-radioactive ceramic-based waste form utilizing a CCIM.

  4. Fundamental Aspects of Zeolite Waste Form Production by Hot Isostatic Pressing

    Jubin, Robert Thomas [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Bruffey, Stephanie H. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Jordan, Jacob A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2017-02-01

    The direct conversion of iodine-bearing sorbents into a stable waste form is a research topic of interest to the US Department of Energy. The removal of volatile radioactive 129I from the off-gas of a nuclear fuel reprocessing facility will be necessary in order to comply with the regulatory requirements that apply to facilities sited within the United States (Jubin et al., 2012a), and any iodine-containing media or solid sorbents generated by this process would contain 129I and would be destined for eventual geological disposal. While recovery of iodine from some sorbents is possible, a method to directly convert iodineloaded sorbents to a durable waste form with little or no additional waste materials being formed and a potentially reduced volume would be beneficial. To this end, recent studies have investigated the conversion of iodine-loaded silver mordenite (I-AgZ) directly to a waste form by hot isostatic pressing (HIPing) (Bruffey and Jubin, 2015). Silver mordenite (AgZ), of the zeolite class of minerals, is under consideration for use in adsorbing iodine from nuclear reprocessing off-gas streams. Direct conversion of I-AgZ by HIPing may provide the following benefits: (1) a waste form of high density that is tolerant to high temperatures, (2) a waste form that is not significantly chemically hazardous, and (3) a robust conversion process that requires no pretreatment.

  5. Identification of major degradation products of 5-aminosalicylic acid formed in aqueous solutions and in pharmaceuticals

    Jensen, J.; Cornett, Claus; Olsen, C. E.

    1992-01-01

    of polymeric species by oxidative self-coupling of 5-ASA moieties. These results indicate that the degradation of 5-ASA follows the same mechanism as observed for the autooxidation of 4-aminophenol and 1,4-phenylenediamine. Some of the identified degradation products were found in 5-ASA......The formation of four major degradation products of 5-aminosalicylic acid (5-ASA) in buffered solutions at pH 7.0 was demonstrated by gradient HPLC analysis. The isolation and structural elucidation of the resulting degradation products showed that the degradation of 5-ASA led to the formation...

  6. Exploiting the aerobic endospore-forming bacterial diversity in saline and hypersaline environments for biosurfactant production.

    de Almeida Couto, Camila Rattes; Alvarez, Vanessa Marques; Marques, Joana Montezano; de Azevedo Jurelevicius, Diogo; Seldin, Lucy

    2015-10-28

    Biosurfactants are surface-active biomolecules with great applicability in the food, pharmaceutical and oil industries. Endospore-forming bacteria, which survive for long periods in harsh environments, are described as biosurfactant producers. Although the ubiquity of endospore-forming bacteria in saline and hypersaline environments is well known, studies on the diversity of the endospore-forming and biosurfactant-producing bacterial genera/species in these habitats are underrepresented. In this study, the structure of endospore-forming bacterial communities in sediment/mud samples from Vermelha Lagoon, Massambaba, Dois Rios and Abraão Beaches (saline environments), as well as the Praia Seca salterns (hypersaline environments) was determined via denaturing gradient gel electrophoresis. Bacterial strains were isolated from these environmental samples and further identified using 16S rRNA gene sequencing. Strains presenting emulsification values higher than 30 % were grouped via BOX-PCR, and the culture supernatants of representative strains were subjected to high temperatures and to the presence of up to 20 % NaCl to test their emulsifying activities in these extreme conditions. Mass spectrometry analysis was used to demonstrate the presence of surfactin. A diverse endospore-forming bacterial community was observed in all environments. The 110 bacterial strains isolated from these environmental samples were molecularly identified as belonging to the genera Bacillus, Thalassobacillus, Halobacillus, Paenibacillus, Fictibacillus and Paenisporosarcina. Fifty-two strains showed emulsification values of at least 30%, and they were grouped into 18 BOX groups. The stability of the emulsification values varied when the culture supernatants of representative strains were subjected to high temperatures and to the presence of up to 20% NaCl. The presence of surfactin was demonstrated in one of the most promising strains. The environments studied can harbor endospore-forming

  7. Production of Curved Precast Concrete Elements for Shell Structures and Free-form Architecture using the Flexible Mould Method

    Schipper, H.R.; Grünewald, S.; Eigenraam, P.; Raghunath, P.; Kok, M.A.D.

    2014-01-01

    Free-form buildings tend to be expensive. By optimizing the production process, economical and well-performing precast concrete structures can be manufactured. In this paper, a method is presented that allows producing highly accurate double curved-elements without the need for milling two expensive

  8. 21 CFR 1240.61 - Mandatory pasteurization for all milk and milk products in final package form intended for direct...

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Mandatory pasteurization for all milk and milk... pasteurization for all milk and milk products in final package form intended for direct human consumption. (a) No... pasteurization are provided for by regulation, such as in part 133 of this chapter for curing of certain cheese...

  9. 30 CFR 210.201 - How do I submit Form MMS-4430, Solid Minerals Production and Royalty Report?

    2010-07-01

    ..., Colorado 80217-5810; or (ii) For courier service or overnight mail (excluding Express Mail): Minerals... 30 Mineral Resources 2 2010-07-01 2010-07-01 false How do I submit Form MMS-4430, Solid Minerals Production and Royalty Report? 210.201 Section 210.201 Mineral Resources MINERALS MANAGEMENT SERVICE...

  10. Analysis of microfouling products formed on metallic surfaces exposed in a marine environment

    DeSouza, F.P.; Bhosle, N.B.

    . The suspended matter and fouling products on the MS, SS and Cu panels were analysed for organic carbon (OC), organic nitrogen (ON), chlorophyll a (chl a), protein and carbohydrate concentration and composition, and the dry weight (DW) was recorded. Compared...

  11. Liquid composition having ammonia borane and decomposing to form hydrogen and liquid reaction product

    Davis, Benjamin L; Rekken, Brian D

    2014-04-01

    Liquid compositions of ammonia borane and a suitably chosen amine borane material were prepared and subjected to conditions suitable for their thermal decomposition in a closed system that resulted in hydrogen and a liquid reaction product.

  12. WATER BOND FORMS IN THE DOUGH AND SORBTION PROPERTIES OF GLUTEN-FREE MACARONI PRODUCTS MADE FROM CORN FLOUR

    Olexandr ROZHNO

    2017-09-01

    Full Text Available Authors have developed macaroni products made from fine meal corn flour by structure forming additives of different nature and without them. The correlation between water of different bond forms in macaroni dough with different structural forming additives and without them has been investigated. Results show that water of macro and microcapillaries is prevalent in the corn dough – 39.40 - 54.69 % of overall amount of water. Osmotically bound water amounts 18.75 – 28.04 %, adsorbically bound water -18.49 – 23.13 % of overall amount of water. The absorbtion capability of the macaroni products and amount of adsorbed water has been determined. The micropore structure of these samples was characterized. The correlation between structures of macaroni products, both amount of adsorbed moisture and energy of sorption were proven. The amount of monomolecular layer’s moisture for gluten-free corn macaroni products is significantly higher – in 1.2 – 1.5 times – when compared to the wheat macaroni products. Due to this fact, corn samples obtained higher energy of moisture sorption. The correlation between structural characteristics of the macaroni samples and their quality was shown.

  13. Size dependence in non-sperm ejaculate production is reflected in daily energy expenditure and resting metabolic rate.

    Friesen, Christopher R; Powers, Donald R; Copenhaver, Paige E; Mason, Robert T

    2015-05-01

    The non-sperm components of an ejaculate, such as copulatory plugs, can be essential to male reproductive success. But the costs of these ejaculate components are often considered trivial. In polyandrous species, males are predicted to increase energy allocation to the production of non-sperm components, but this allocation is often condition dependent and the energetic costs of their production have never been quantified. Red-sided garter snakes (Thamnophis sirtalis parietalis) are an excellent model with which to quantify the energetic costs of non-sperm components of the ejaculate as they exhibit a dissociated reproductive pattern in which sperm production is temporally disjunct from copulatory plug production, mating and plug deposition. We estimated the daily energy expenditure and resting metabolic rate of males after courtship and mating, and used bomb calorimetry to estimate the energy content of copulatory plugs. We found that both daily energy expenditure and resting metabolic rate were significantly higher in small mating males than in courting males, and a single copulatory plug without sperm constitutes 5-18% of daily energy expenditure. To our knowledge, this is the first study to quantify the energetic expense of size-dependent ejaculate strategies in any species. © 2015. Published by The Company of Biologists Ltd.

  14. PROCESS FOR THE PRODUCTION OF AN ACTIVATED FORM OF UO$sub 2$

    Polissar, M.J.

    1957-09-24

    A process for producing a highly active form of UO/sub 2/ characterized both by rapid oxidation in air and by rapid chlorination with CCl/sub 4/ vapor at an elevated temperature is reported. In accordance with the process, commercial UO/sub 2/, is subjected to a series of oxidation-reduction operations to produce a form of UC/sub 2/ of enhanced reactivity. By treatimg commercial UO/sub 2/ at a temperature between 335 and 485 deg C with methane, then briefly with an oxygen containing gas and followimg this by a second treatment with a methane containing gas, the original relatively stable charge of UO/sub 2/ will be transformed into an active form of UO/sub 2/.

  15. Impact of CHO Metabolism on Cell Growth and Protein Production: An Overview of Toxic and Inhibiting Metabolites and Nutrients

    Pereira, Sara; Kildegaard, Helene F.; Andersen, Mikael R.

    2018-01-01

    and process optimization and monitoring to perform efficiently. One of the main reasons for this is the production and accumulation of toxic and growth-inhibiting metabolites during culture. Lactate and ammonium are the most known, but many more have been identified. In this review, we present an overview...... of metabolites that deplete and accumulate throughout the course of cultivations with toxic and growth inhibitory effects to the cells. We further provide an overview of the CHO metabolism with emphasis to metabolic pathways of amino acids, glutathione (GSH), and related compounds which have growth...... of resources that describe the cellular mechanisms of CHO and are available on-line. Finally, we discuss the application of this knowledge for bioprocess and medium development and cell line engineering....

  16. Euglena in time: Evolution, control of central metabolic processes and multi-domain proteins in carbohydrate and natural product biochemistry

    Ellis C. O’Neill

    2015-12-01

    Full Text Available Euglena gracilis is a eukaryotic microalgae that has been the subject of scientific study for hundreds of years. It has a complex evolutionary history, with traces of at least four endosymbiotic genomes and extensive horizontal gene transfer. Given the importance of Euglena in terms of evolutionary cell biology and its unique taxonomic position, we initiated a de novo transcriptome sequencing project in order to understand this intriguing organism. By analysing the proteins encoded in this transcriptome, we can identify an extremely complex metabolic capacity, rivalling that of multicellular organisms. Many genes have been acquired from what are now very distantly related species. Herein we consider the biology of Euglena in different time frames, from evolution through control of cell biology to metabolic processes associated with carbohydrate and natural products biochemistry.

  17. Calculation of scalar products of wave functions and form factors in the framework of the algebraic Bethe ansatz

    Slavnov, N.A.

    1989-01-01

    The Bethe ansatz method is widely used to investigate two-dimensional completely integrable models. In the framework of the quantum inverse scattering method it has proved to be possible to construct an algebraic scheme of the Bethe ansatz, and this has been successfully applied to calculation of correlation functions. One of the important questions of the method is that of the scalar products of the wave functions. In particular, knowledge of the properties of the scalar products is necessary for investigating the form factors and correlation function. In the present paper the author considers a generalized model with R matrix of the model of the nonlinear Schroedinger equation. The main formulas and notation are given in Sec. 2. In Sec. 3 he calculates the scalar product of an arbitrary function and an eigenfunction of the Hamiltonian. The generalized two-site model is introduced in Sec. 4. In Sec. 5 he calculates the form factor of the particle number operator

  18. Alkaline conditions stimulate the production of 1,3-propanediol in Lactobacillus panis PM1 through shifting metabolic pathways.

    Grahame, Douglas A S; Kang, Tae Sun; Khan, Nurul H; Tanaka, Takuji

    2013-07-01

    A novel Lactobacillus panis PM1 isolate was found to be capable of converting glycerol to 1,3-propanediol (1,3-PDO), an increasingly valuable commodity chemical. In this study the effects of various process parameters, including glucose and glycerol concentrations, inoculum size, temperature, aeration, pH, and carbon source were examined to determine the optimal conditions for the production of 1,3-PDO using a culture method simulating late log to early stationary phases. Inoculum size did not influence the production of 1,3-PDO, and temperature variance showed similar 1,3-PDO production between 25 and 37 °C under the examined conditions. Glycerol concentration and pH played a primary role in the final concentration of 1,3-PDO. The highest production occurred at 150-250 mM glycerol when 50 mM glucose was available. Alkaline initial conditions (pH 9-10) stimulated the production of 1,3-PDO which concurrently occurred with increased acetic acid production. Under these conditions, 213.6 mM of 1,3-PDO were produced from 300 mM glycerol (conversion efficiency was 71 %). These observations indicated that the production of 1,3-PDO was associated with the shift of the metabolic end-product ethanol to acetic acid, and that this shift resulted in an excess concentration of NADH available for the processing of glycerol to 1,3-PDO.

  19. Metabolic engineering of the pentose phosphate pathway for enhanced limonene production in the cyanobacterium Synechocysti s sp. PCC 6803.

    Lin, Po-Cheng; Saha, Rajib; Zhang, Fuzhong; Pakrasi, Himadri B

    2017-12-13

    Isoprenoids are diverse natural compounds, which have various applications as pharmaceuticals, fragrances, and solvents. The low yield of isoprenoids in plants makes them difficult for cost-effective production, and chemical synthesis of complex isoprenoids is impractical. Microbial production of isoprenoids has been considered as a promising approach to increase the yield. In this study, we engineered the model cyanobacterium Synechocystis sp. PCC 6803 for sustainable production of a commercially valuable isoprenoid, limonene. Limonene synthases from the plants Mentha spicata and Citrus limon were expressed in cyanobacteria for limonene production. Production of limonene was two-fold higher with limonene synthase from M. spicata than that from C. limon. To enhance isoprenoid production, computational strain design was conducted by applying the OptForce strain design algorithm on Synechocystis 6803. Based on the metabolic interventions suggested by this algorithm, genes (ribose 5-phosphate isomerase and ribulose 5-phosphate 3-epimerase) in the pentose phosphate pathway were overexpressed, and a geranyl diphosphate synthase from the plant Abies grandis was expressed to optimize the limonene biosynthetic pathway. The optimized strain produced 6.7 mg/L of limonene, a 2.3-fold improvement in productivity. Thus, this study presents a feasible strategy to engineer cyanobacteria for photosynthetic production of isoprenoids.

  20. Heme metabolism in stress regulation and protein production: from Cinderella to a key player

    Martinez Ruiz, José Luis; Petranovic, D.; Nielsen, Jens

    2016-01-01

    Heme biosynthesis is a highly conserved pathway which is present in all kingdoms, from Archaea to higher organisms such as plants and mammals. The heme molecule acts as a prosthetic group for different proteins and enzymes involved in energy metabolism and reactions involved in electron transfer....

  1. From physiology to systems metabolic engineering for the production of biochemicals by lactic acid bacteria

    Gaspar, Paula; Carvalho, Ana L.; Vinga, Susana

    2013-01-01

    of comprehensive data on their metabolism and genetics was generated throughout the years. This knowledge has been instrumental in the implementation of successful applications in the food industry, such as the selection of robust starter cultures with desired phenotypic traits. The advent of genomics, functional...

  2. Seasonal and interannual patterns in primary production, respiration and net ecosystem metabolism in three estuaries in the northeast Gulf of Mexico

    Measurements of primary production and respiration provide fundamental information about the trophic status of aquatic ecosystems, yet such measurements are logistically difficult and expensive to sustain as part of long-term monitoring programs. However, ecosystem metabolism par...

  3. Method for forming nuclear fuel containers of a composite construction and the product thereof

    Cheng, B.-C.; Rosenbaum, H.S.; Armijo, J.S.

    1981-01-01

    An improved method of producing a composite nuclear fuel container is described which comprises a casing or fuel sheath of zirconium or its alloy with a lining cladding of deposited copper superimposed over the inside surface of the zirconium or alloy and a layer of oxide of the zirconium or alloy formed on the inside surface of the casing or sheath. (U.K.)

  4. Friction modelling in sheet metal forming simulations: application and validation on an U-Bend product

    Sigvant, Mats; Hol, Johan; Chezan, Toni; van den Boogaard, Ton; Hora, P.

    2015-01-01

    The accuracy of sheet metal forming simulations strongly depends on, amongst others, friction modelling. The industrial standard is to use the Coulomb friction model with a constant coefficient of friction. However, it is known that the true friction conditions are dependent on the tribology system,

  5. Non-cytochrome mediated mitochondrial ATP production in bloodstream form Trypanosoma brucei brucei

    Bienen, E. J.; Maturi, R. K.; Pollakis, G.; Clarkson, A. B.

    1993-01-01

    The life cycle of Trypanosoma brucei brucei involves a series of differentiation steps characterized by marked changes in mitochondrial development and function. The bloodstream forms of this parasite completely lack cytochromes and have not been considered to have any Krebs cycle function. It has

  6. Representation of National Identity in Malaysian State Mosque Built Form as a Socio- cultural Product

    Alice Sabrina Ismail

    2018-01-01

    Full Text Available The formation of national identity is the essence for a multi-racial country like Malaysia because it can strengthen national solidarity, create a common aspiration, and sustain the continuity of a historical heritage with multiple ethnicity and background. Nevertheless, the characteristics of this national identity is still not fully dealt with from an architectural aspect. Hence, the purpose of this paper is to critically examine the problem of national architectural identity in Malaysia. This is vital to uncover the resulting design principles with national architectural identity characteristics by referencing to three typologies of local state mosque as case study. The finding outlines that there are six architectural principles that influence the formation of national identity. These are the understanding of built form detail elements, materials usage, local style composition and expression, application of organic theory, designing a regionalistic articulation of space and form as well as an understanding of democratic values form to reflect the characteristics of national identity. These established referential guideline design on national identity is of benefit for future designers, builders, developer and related authority to produce built form that symbolizes nation political values as well as responsive to the existing social culture context.

  7. Methane production and digestion of different physical forms of rapeseed as fat supplements in dairy cows

    Brask, Maike; Lund, Peter; Weisbjerg, Martin Riis

    2013-01-01

    The purpose of this experiment was to study the effect of the physical form of rapeseed fat on methane (CH4) mitigation properties, feed digestion, and rumen fermentation. Four lactating ruminal-, duodenal-, and ileal-cannulated Danish Holstein dairy cows (143 d in milk, milk yield of 34.3 kg) were...

  8. Particle production and Boltzmann integral form of relativistic quantum transport theory

    Rafelski, J.; Davis, E.D.; Bialynicki-Birula, I.

    1993-01-01

    The 3+3+1 dimensional relativistic quantum transport equation for the fermion matter field, combines the particle pair production with flow phenomena, which occur at very different time scale. A direct numerical treatment of dynamical situations is therefore practically impossible. The authors attempt a seperation of these two sectors by the method of prediagonalization of the integral equations. They exploit the structure of the resolvent of the transport equations: it contains two poles corresponding to the flow sector and two to the pair production sector. Their hope for practical applications is to treat matter flow as a classical phenomenon and to be able to obtain an integral term describing the pair production accurately

  9. Origin of Nanobubbles Electrochemically Formed in a Magnetic Field: Ionic Vacancy Production in Electrode Reaction

    Aogaki, Ryoichi; Sugiyama, Atsushi; Miura, Makoto; Oshikiri, Yoshinobu; Miura, Miki; Morimoto, Ryoichi; Takagi, Satoshi; Mogi, Iwao; Yamauchi, Yusuke

    2016-07-01

    As a process complementing conventional electrode reactions, ionic vacancy production in electrode reaction was theoretically examined; whether reaction is anodic or cathodic, based on the momentum conservation by Newton’s second law of motion, electron transfer necessarily leads to the emission of original embryo vacancies, and dielectric polarization endows to them the same electric charge as trans- ferred in the reaction. Then, the emitted embryo vacancies immediately receive the thermal relaxation of solution particles to develop steady-state vacancies. After the vacancy production, nanobubbles are created by the collision of the vacancies in a vertical magnetic field.

  10. Waste acceptance product specifications for vitrified high-level waste forms

    Applewhite-Ramsey, A.; Sproull, J.F.

    1994-01-01

    The Department of Energy (DOE) Office of Environmental Restoration and Waste Management (EM) has developed Waste Acceptance Product Specifications (EM-WAPS). The EM-WAPS will be the basis for defining product acceptance criteria compatible with the requirements of the Civilian Radioactive Waste Management System (CRWMS). The relationship between the EM-WAPS and the CRWMS Systems Requirements document (WA-SRD) will be discussed. The impact of the EM-WAPS on the Savannah River Sit (SRS) Defense Waste Processing Facility's (DWPF) Waste Acceptance Program, Waste Qualification Run planning, and startup schedule will also be reported. 14 refs., 2 tabs

  11. Beneath the veil: Plant growth form influences the strength of species richness-productivity relationships in forests

    Oberle, B.; Grace, J.B.; Chase, J.M.

    2009-01-01

    Aim: Species richness has been observed to increase with productivity at large spatial scales, though the strength of this relationship varies among functional groups. In forests, canopy trees shade understorey plants, and for this reason we hypothesize that species richness of canopy trees will depend on macroclimate, while species richness of shorter growth forms will additionally be affected by shading from the canopy. In this study we test for differences in species richness-productivity relationships (SRPRs) among growth forms (canopy trees, shrubs, herbaceous species) in small forest plots. Location: We analysed 231 plots ranging from 34.0?? to 48.3?? N latitude and from 75.0?? to 124.2?? W longitude in the United States. Methods: We analysed data collected by the USDA Forest Inventory and Analysis program for plant species richness partitioned into different growth forms, in small plots. We used actual evapotranspiration as a macroclimatic estimate of regional productivity and calculated the area of light-blocking tissue in the immediate area surrounding plots for an estimate of the intensity of local shading. We estimated and compared SRPRs for different partitions of the species richness dataset using generalized linear models and we incorporated the possible indirect effects of shading using a structural equation model. Results: Canopy tree species richness increased strongly with regional productivity, while local shading primarily explained the variation in herbaceous plant richness. Shrub species richness was related to both regional productivity and local shading. Main conclusions: The relationship between total forest plant species richness and productivity at large scales belies strong effects of local interactions. Counter to the pattern for overall richness, we found that understorey herbaceous plant species richness does not respond to regional productivity gradients, and instead is strongly influenced by canopy density, while shrub species

  12. Advances in metabolic pathway and strain engineering paving the way for sustainable production of chemical building blocks.

    Chen, Yun; Nielsen, Jens

    2013-12-01

    Bio-based production of chemical building blocks from renewable resources is an attractive alternative to petroleum-based platform chemicals. Metabolic pathway and strain engineering is the key element in constructing robust microbial chemical factories within the constraints of cost effective production. Here we discuss how the development of computational algorithms, novel modules and methods, omics-based techniques combined with modeling refinement are enabling reduction in development time and thus advance the field of industrial biotechnology. We further discuss how recent technological developments contribute to the development of novel cell factories for the production of the building block chemicals: adipic acid, succinic acid and 3-hydroxypropionic acid. Copyright © 2013 Elsevier Ltd. All rights reserved.

  13. Introduction to metabolic genetic engineering for the production of valuable secondary metabolites in in vivo and in vitro plant systems.

    Benedito, Vagner A; Modolo, Luzia V

    2014-01-01

    Plants are capable of producing a myriad of chemical compounds. While these compounds serve specific functions in the plant, many have surprising effects on the human body, often with positive action against diseases. These compounds are often difficult to synthesize ex vivo and require the coordinated and compartmentalized action of enzymes in living organisms. However, the amounts produced in whole plants are often small and restricted to single tissues of the plant or even cellular organelles, making their extraction an expensive process. Since most natural products used in therapeutics are specialized, secondary plant metabolites, we provide here an overview of the classification of the main classes of these compounds, with its biochemical pathways and how this information can be used to create efficient in and ex planta production pipelines to generate highly valuable compounds. Metabolic genetic engineering is introduced in light of physiological and genetic methods to enhance production of high-value plant secondary metabolites.

  14. Amino acid and glucose metabolism in fed-batch CHO cell culture affects antibody production and glycosylation

    Fan, Yuzhou; Jimenez Del Val, Ioscani; Müller, Christian

    2015-01-01

    optimization, especially media optimization. Gaining knowledge on their interrelations could provide insight for obtaining higher immunoglobulin G (IgG) titer and better controlling glycosylationrelated product quality. In this work, different fed-batch processes with two chemically defined proprietary media......Fed-batch Chinese hamster ovary (CHO) cell culture is the most commonly used process for IgG production in the biopharmaceutical industry. Amino acid and glucose consumption, cell growth, metabolism, antibody titer, and N-glycosylation patterns are always the major concerns during upstream process...... and glutamine concentrations and uptake rates were positively correlated with intracellular UDP-Gal availability. All these findings are important for optimization of fed-batch culture for improving IgG production and directing glycosylation quality....

  15. Warming affects growth rates and microcystin production in tropical bloom-forming microcystis strains

    Bui, Trung; Dao, Thanh Son; Vo, Truong Giang; Lürling, Miquel

    2018-01-01

    Warming climate is predicted to promote cyanobacterial blooms but the toxicity of cyanobacteria under global warming is less well studied. We tested the hypothesis that raising temperature may lead to increased growth rates but to decreased microcystin (MC) production in tropical Microcystis

  16. Forming the face of green products : Mood boards and early consumer involvement in ship interior design

    Murto, P.; Ahola, M.; Person, F.E.O.K.

    2013-01-01

    In this paper, we study mood boards and the process by which they can be used to understand consumer inferences about environmental sustainability in the early stages of green product development. As a component of a research project on sustainable ship design, we created mood boards showcasing

  17. The Role of Geminates in Infants' Early Word Production and Word-Form Recognition

    Vihman, Marilyn; Majoran, Marinella

    2017-01-01

    Infants learning languages with long consonants, or geminates, have been found to "overselect" and "overproduce" these consonants in early words and also to commonly omit the word-initial consonant. A production study with thirty Italian children recorded at 1;3 and 1;9 strongly confirmed both of these tendencies. To test the…

  18. Tracing biofouling to the structure of the microbial community and its metabolic products: a study of the three-stage MBR process.

    Gao, Dawen; Fu, Yuan; Ren, Nanqi

    2013-11-01

    The biofouling characteristics of a sequential anoxic/aerobic-membrane bioreactor (A/O MBR) were analyzed during the three-stage process (fast-slow-fast transmembrane pressure (TMP) increasing). The results indicated: during the stage 1 (before day 1), the microbial communities in the activated sludge (AS), cake sludge (CS) and biofilm (BF) were similar to each other, and the adsorption of microbes and the metabolic products was the main factor that led to TMP increase; during the stage 2 (between day 1 and day 7), the cake layer begun to form and the TMP continued to rise gradually at a reduced rate compared to stage 1, at this point a characteristic microbial community colonized the CS with microorganisms such as Saprospiraceae and Comamonadaceae thriving on the membrane surface (BF) probably due to greater nutrient availability, and the predominance of these species in the microbial population led to the accumulation of biofouling metabolic products in the CS, which resulted in membrane fouling and the associated rise in TMP; during the final stage (after day 7), the biofilm had matured, and the activity of anaerobes stimulated cake compaction. The statistical analysis showed a correlation between the TMP changing rate and the carbonhydrates of soluble microbial products (SMPc) content in the CS. When the SMPc concentration rose slowly there was a low level of biofouling. However, when the SMPc accumulating rate was greater, it resulted in the more severe biofouling associated with the TMP jump. Furthermore, the correlation coefficient for the TMP increase and protein concentrations of extracellular polymeric substances (EPSp) in the CS was highly significant. The cluster analysis suggested that the AS microbial community remained stable during the three TMP change stages, while the CS and BF community were changed accompanied with the TMP change. The interaction between the microbial communities and the metabolic products lead to the significant correlation

  19. Post-fermentative production of glutathione by baker's yeast (S. cerevisiae) in compressed and dried forms.

    Musatti, Alida; Manzoni, Matilde; Rollini, Manuela

    2013-01-25

    The study was aimed at investigating the best biotransformation conditions to increase intracellular glutathione (GSH) levels in samples of baker's yeast (Saccharomyces cerevisiae) employing either the commercially available compressed and dried forms. Glucose, GSH precursors amino acids, as well as other cofactors, were dissolved in a biotransformation solution and yeast cells were added (5%dcw). Two response surface central composite designs (RSCCDs) were performed in sequence: in the first step the influence of amino acid composition (cysteine, glycine, glutamic acid and serine) on GSH accumulation was investigated; once their formulation was set up, the influence of other components was studied. Initial GSH content was found 0.53 and 0.47%dcw for compressed and dried forms. GSH accumulation ability of baker's yeast in compressed form was higher at the beginning of shelf life, that is, in the first week, and a maximum of 2.04%dcw was obtained. Performance of yeast in dried form was not found satisfactory, as the maximum GSH level was 1.18%dcw. When cysteine lacks from the reaction solution, yeast cells do not accumulate GSH. With dried yeast, the highest GSH yields occurred when cysteine was set at 3 g/L, glycine and glutamic acid at least at 4 g/L, without serine. Employing compressed yeast, the highest GSH yields occurred when cysteine and glutamic acid were set at 2-3 g/L, while glycine and serine higher than 2 g/L. Results allowed to set up an optimal and feasible procedure to obtain GSH-enriched yeast biomass, with up to threefold increase with respect to initial content. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Research Note--Should Consumers Use the Halo to Form Product Evaluations?

    Peter Boatwright; Ajay Kalra; Wei Zhang

    2008-01-01

    In purchase situations where attribute information is either missing or difficult to judge, a well-known heuristic that consumers use to form evaluations is the halo effect. The psychology literature has widely considered the halo a reflection of consumers' inability to discriminate between different attributes and have therefore labeled it the "halo error" or the "logical error." The objective of this paper is to offer a rationale for the halo effect. We use a decision-theory framework to sh...

  1. On determinant representations of scalar products and form factors in the SoV approach: the XXX case

    Kitanine, N.; Maillet, J. M.; Niccoli, G.; Terras, V.

    2016-03-01

    In the present article we study the form factors of quantum integrable lattice models solvable by the separation of variables (SoVs) method. It was recently shown that these models admit universal determinant representations for the scalar products of the so-called separate states (a class which includes in particular all the eigenstates of the transfer matrix). These results permit to obtain simple expressions for the matrix elements of local operators (form factors). However, these representations have been obtained up to now only for the completely inhomogeneous versions of the lattice models considered. In this article we give a simple algebraic procedure to rewrite the scalar products (and hence the form factors) for the SoV related models as Izergin or Slavnov type determinants. This new form leads to simple expressions for the form factors in the homogeneous and thermodynamic limits. To make the presentation of our method clear, we have chosen to explain it first for the simple case of the XXX Heisenberg chain with anti-periodic boundary conditions. We would nevertheless like to stress that the approach presented in this article applies as well to a wide range of models solved in the SoV framework.

  2. On determinant representations of scalar products and form factors in the SoV approach: the XXX case

    Kitanine, N; Maillet, J M; Niccoli, G; Terras, V

    2016-01-01

    In the present article we study the form factors of quantum integrable lattice models solvable by the separation of variables (SoVs) method. It was recently shown that these models admit universal determinant representations for the scalar products of the so-called separate states (a class which includes in particular all the eigenstates of the transfer matrix). These results permit to obtain simple expressions for the matrix elements of local operators (form factors). However, these representations have been obtained up to now only for the completely inhomogeneous versions of the lattice models considered. In this article we give a simple algebraic procedure to rewrite the scalar products (and hence the form factors) for the SoV related models as Izergin or Slavnov type determinants. This new form leads to simple expressions for the form factors in the homogeneous and thermodynamic limits. To make the presentation of our method clear, we have chosen to explain it first for the simple case of the XXX Heisenberg chain with anti-periodic boundary conditions. We would nevertheless like to stress that the approach presented in this article applies as well to a wide range of models solved in the SoV framework. (paper)

  3. Sustainable source of omega-3 eicosapentaenoic acid from metabolically engineered Yarrowia lipolytica: from fundamental research to commercial production.

    Xie, Dongming; Jackson, Ethel N; Zhu, Quinn

    2015-02-01

    The omega-3 fatty acids, cis-5, 8, 11, 14, and 17-eicosapentaenoic acid (C20:5; EPA) and cis-4, 7, 10, 13, 16, and 19-docosahexaenoic acid (C22:6; DHA), have wide-ranging benefits in improving heart health, immune function, mental health, and infant cognitive development. Currently, the major source for EPA and DHA is from fish oil, and a minor source of DHA is from microalgae. With the increased demand for EPA and DHA, DuPont has developed a clean and sustainable source of the omega-3 fatty acid EPA through fermentation using metabolically engineered strains of Yarrowia lipolytica. In this mini-review, we will focus on DuPont's technology for EPA production. Specifically, EPA biosynthetic and supporting pathways have been introduced into the oleaginous yeast to synthesize and accumulate EPA under fermentation conditions. This Yarrowia platform can also produce tailored omega-3 (EPA, DHA) and/or omega-6 (ARA, GLA) fatty acid mixtures in the cellular lipid profiles. Fundamental research such as metabolic engineering for strain construction, high-throughput screening for strain selection, fermentation process development, and process scale-up were all needed to achieve the high levels of EPA titer, rate, and yield required for commercial application. Here, we summarize how we have combined the fundamental bioscience and the industrial engineering skills to achieve large-scale production of Yarrowia biomass containing high amounts of EPA, which led to two commercial products, New Harvest™ EPA oil and Verlasso® salmon.

  4. Contributions of citrate in redox potential maintenance and ATP production: metabolic pathways and their regulation in Lactobacillus panis PM1.

    Kang, Tae Sun; Korber, Darren R; Tanaka, Takuji

    2013-10-01

    Lactobacillus panis PM1 belongs to the group III heterofermentative lactobacilli and can utilize various NADH-reoxidizing routes (e.g., citrate, glycerol, and oxygen) according to environmental conditions. In this study, we investigated the ability of L. panis PM1 to produce succinate, acetate, and lactate via citrate utilization. Possible pathways, as well as regulation, for citrate metabolism were examined on the basis of the genome sequence data and metabolic profiles of L. panis PM1. The presence of citrate led to the up-regulation, at the transcriptional level, of the genes encoding for citrate lyase, malate dehydrogenase, and malic enzyme of the citrate pathways by 10- to 120-fold. The transcriptional regulator of the dha operon coding for glycerol dehydratase of L. panis PM1 repressed the expression of the citrate lyase gene (10-fold). Metabolite analyses indicated that the transcriptional enhancement by citrate stimulated succinate yield. Citrate metabolism contributed to energy production by providing a major alternate pathway for NAD(+) regeneration and allowed acetyl phosphate to yield acetate/ATP instead of ethanol/NAD(+). Additionally, a branching pathway from oxaloacetate to pyruvate increased the pool of lactate, which was then used to produce ATP during stationary phase. However, the redirection of NADH-to-citrate utilization resulted in stress caused by end-products (i.e., succinate and acetate). This stress reduced succinate production by up to 50 % but did not cause significant changes at transcriptional level. Overall, citrate utilization was beneficial for the growth of L. panis PM1 by providing a NAD(+) regeneration route and producing extra ATP.

  5. Characterization of the corrosion products formed on mild steel in acidic medium with N-octadecylpyridinium bromide as corrosion inhibitor

    Nava, N., E-mail: tnava@imp.mx; Likhanova, N. V. [Direccion de Investigacion y Posgrado, Instituto Mexicano del Petroleo (Mexico); Olivares-Xometl, O. [Benemerita Universidad Autonoma de Puebla, Facultad de Ingenieria Quimica (Mexico); Flores, E. A. [Direccion de Investigacion y Posgrado, Instituto Mexicano del Petroleo (Mexico); Lijanova, I. V. [CIITEC, Instituto Politecnico Nacional (Mexico)

    2011-11-15

    The characterization of the corrosion products formed on mild steel SAE 1018 after 2 months exposure in aqueous sulfuric acid with and without corrosion inhibitor N-octadecylpyridinium bromide has been carried out by means of transmission {sup 57}Fe Moessbauer spectroscopy and X-ray powder diffraction (XRD). The major constituent of the rust formed in this environment without corrosion inhibitor is goethite ({alpha}-FeOOH). The samples with N-octadecylpyridinium bromide contain rozenite and large amounts of melanterite in the corrosion layers.

  6. Assesment of systemic exposure form a wound contaminated by radioactive products

    Piechowski, Jean; Menoux, Bernadette; Chaptinel, Yves

    1992-03-01

    Wound contamination may occur in case of accidental exposure of workers handling radioactive products. In order to operate radio-toxicological follow-up, a fairly general model must be able to apply to the various injuries and products involved. Some principles and their applications are developed to make it possible for physicians to set up a monitoring program relevant to the assessment of the systemic burden. Only the dosimetry relative to the activity passing through the cutaneous mucous barrier into the blood flow is considered. Local doses at the wound level are not considered. For the interpretation of the measurements, both retention and excretion curves and dosimetric factors are given for the most common radionuclides. (authors) [fr

  7. THE CONCEPTUAL FRAMEWORK OF FORMING THE PRODUCT QUALITY CONTROL STRATEGY OF ROAD-BUILDING ENGINEERING ENTERPRISES

    Mariia Sadova

    2017-11-01

    Full Text Available The purpose of the work is the need of the theoretical justification and the development of relevant scientific and methodological approaches to the management of the road production quality of machine-building enterprises; it led to the choice of research topics. Methodology. Taking into account the value of existing methods of quality management of road production engineering companies, currently a particular attention requires improvement of the quality control functions: analysis, evaluation, planning and quality control during the operation of road-building equipment, because existing economic practices and quality management approaches are built on classical principles of ISO 9001, and in the practical application do not solve all of the drawbacks of traditional management models, but only smooth them a little. Therefore, the formation of the quality management strategy for the road production of mechanical engineering is essential to ensure the competitiveness of products and enterprises of the road-building complex in the period of development of market relations in the economy. The results showed that the current market conditions impose strict requirements to all production areas of the society. Especially stringent are requirements for the road construction, because the quality of the construction of the subgrade, covering roads and curbs, grading and soil compaction under different conditions of terrain and driving safety depends on people's lives. This factor determines the exceptional quality requirements for roadbuilding equipment, which must be reliable, functional and able to meet the totality of the design and operational characteristics of the road. Further European integration of Ukraine, deepening of external economic relations with the European countries are responsible for the growing needs of the national economy in the development and expansion of the transport network. However, large-scale road construction and repair

  8. Hydrogen production from nano-porous Si powder formed by stain etching

    Litvinenko, S.; Alekseev, S.; Kuznetsov, G.; Skryshevsky, V. [Institute of High Technology of National Taras Shevchenko University of Kyiv, Volodymyrs' ka 64, Kyiv 01601 (Ukraine); Lysenko, V.; Barbier, D. [Lyon Institute of Nanotechnologies (INL), CNRS UMR-5270, University of Lyon, INSA de Lyon, 7 avenue Jean Capelle, Bat. Blaise Pascal, 69621 Villeurbanne Cedex (France); Venturello, A.; Geobaldo, F.; Garrone, E. [Politecnico di Torino, Department of Materials Science and Chemical Engineering, 10129 Torino (Italy); Gulina, L.; Tolstoy, V. [St-Petersburg State University, Chemical Department (Russian Federation)

    2010-07-15

    Hydrogen reservoirs based on porous silicon (PS) nanostructures are considered. Silicon-based hydrogen tanks are believed to be applicable for portable device energy supply and compatible with micro-sources of energy of new generation. Stain etching of silicon powder to produce PS is studied as a technology alternative to conventional electrochemical etching and application of the PS powder for hydrogen production is also described. Size selection of initial Si micro-particles constituting the powders was carried out by sedimentation technique. Hydrogen content in PS was investigated by FTIR spectroscopy. Extraction of hydrogen in water environment in presence of small amount of NH{sub 3} as catalyst was shown to have advantages such as safety and tunability, additional production of hydrogen from water dissociation, and a possibility to characterize PS as a hydrogen source material in terms of hydrogen effective shell and crystalline core conception. (author)

  9. Metabolic safety-margins do not differ between cows of high and low genetic merit for milk production

    Knight, Christopher Harold; Alamer, Mohammed A; Sorensen, Annette

    2004-01-01

    Three galactopoietic stimuli, frequent milking (4X), bovine somatotrophin (bST) and thyroxine (T4) were used in an additive stair-step design to achieve maximum output (metabolic capacity) in six peak-lactation cows of high genetic merit (HT) and six of low genetic merit (LT). A further six of ea...... the commonly held belief that selective breeding of dairy cows for high milk production has rendered them markedly more susceptible to metabolic disturbances.......Three galactopoietic stimuli, frequent milking (4X), bovine somatotrophin (bST) and thyroxine (T4) were used in an additive stair-step design to achieve maximum output (metabolic capacity) in six peak-lactation cows of high genetic merit (HT) and six of low genetic merit (LT). A further six of each...... elevated heart rate and significant loss of body weight and condition compared with the combination of 4X and bST. As a result, treatments were discontinued, on an individual cow basis, before completion of this 6-week phase. Time on experiment did not differ between HT and LT. The results do not support...

  10. Okara, a By-Product of Tofu Manufacturing, Modifies Triglyceride Metabolism at the Intestinal and Hepatic Levels.

    Nagata, Yasuo; Yamasaki, Shiho; Torisu, Norihiro; Suzuki, Taishi; Shimamoto, Saya; Tamaru, Shizuka; Tanaka, Kazunari

    2016-01-01

    Irrespective of a well-known hypocholesterolemic action, a few studies have shown a hypotriglyceridemic potential of okara, a by-product of tofu manufacturing. Okara was fed to rats at the level of 2.5 and 5.0% as dietary protein for 4 wk, and serum and hepatic lipid levels were determined. In addition, soy flour, which has a well-known hypolipidemic action, was used to compare effects on lipid metabolism. Mechanisms of action were further evaluated by measuring hepatic enzyme activity, gene expression of lipid metabolism-related proteins and fecal excretion of lipids. Feeding the okara diets resulted in a significantly lower weight of the liver and adipose tissue in a dose-dependent manner. Serum triglyceride levels were more than 50% lower in rats fed the okara diets compared to those fed the control diet. Enzyme activities of fatty acid synthesis were significantly lowered by the okara diet. Fecal weight was significantly higher in the okara group than in the control group, and fecal excretion of steroids tended to be higher. Therefore, a relatively low amount of okara may exert hypotriglyceridemic action in rats in part through decreased hepatic triglyceride synthesis. The present study also suggests an involvement of intestinal events in altered lipid metabolism in rats fed the okara diets.

  11. Toward Higher QA: From Parametric Release of Sterile Parenteral Products to PAT for Other Pharmaceutical Dosage Forms.

    Hock, Sia Chong; Constance, Neo Xue Rui; Wah, Chan Lai

    2012-01-01

    Pharmaceutical products are generally subjected to end-product batch testing as a means of quality control. Due to the inherent limitations of conventional batch testing, this is not the most ideal approach for determining the pharmaceutical quality of the finished dosage form. In the case of terminally sterilized parenteral products, the limitations of conventional batch testing have been successfully addressed with the application of parametric release (the release of a product based on control of process parameters instead of batch sterility testing at the end of the manufacturing process). Consequently, there has been an increasing interest in applying parametric release to other pharmaceutical dosage forms, beyond terminally sterilized parenteral products. For parametric release to be possible, manufacturers must be capable of designing quality into the product, monitoring the manufacturing processes, and controlling the quality of intermediates and finished products in real-time. Process analytical technology (PAT) has been thought to be capable of contributing to these prerequisites. It is believed that the appropriate use of PAT tools can eventually lead to the possibility of real-time release of other pharmaceutical dosage forms, by-passing the need for end-product batch testing. Hence, this literature review attempts to present the basic principles of PAT, introduce the various PAT tools that are currently available, present their recent applications to pharmaceutical processing, and explain the potential benefits that PAT can bring to conventional ways of processing and quality assurance of pharmaceutical products. Last but not least, current regulations governing the use of PAT and the manufacturing challenges associated with PAT implementation are also discussed. Pharmaceutical products are generally subjected to end-product batch testing as a means of quality control. Due to the inherent limitations of conventional batch testing, this is not the most

  12. A Moessbauer and Electrochemical Characterization of the Corrosion Products Formed from Marine and Marine-Antartic Environments

    Ohanian, M.; Caraballo, R.; Dalchiele, E. A.; Quagliata, E. [Instituto de Ingenieria Quimica, Facultad de Ingenieria (Uruguay)

    2003-06-15

    Corrosion products formed on low alloy steel under two marine environments are characterised. Both environments are classified as C4 according to the ISO 9223 Standard. The corrosion products are identified and their relative proportion is determined by Moessbauer spectroscopy (transmission geometry). Free potentials of corrosion are measured to evaluate the activity of their surfaces. Structural characterisation by XRD were performed on selected samples. It is concluded that the principal phases are goethite, lepidocrocite, ferrihidrite and maghemite. The relative amount of each of them changes with time and with the atmospheric dynamics of each environment.

  13. A Moessbauer and Electrochemical Characterization of the Corrosion Products Formed from Marine and Marine-Antartic Environments

    Ohanian, M.; Caraballo, R.; Dalchiele, E. A.; Quagliata, E.

    2003-01-01

    Corrosion products formed on low alloy steel under two marine environments are characterised. Both environments are classified as C4 according to the ISO 9223 Standard. The corrosion products are identified and their relative proportion is determined by Moessbauer spectroscopy (transmission geometry). Free potentials of corrosion are measured to evaluate the activity of their surfaces. Structural characterisation by XRD were performed on selected samples. It is concluded that the principal phases are goethite, lepidocrocite, ferrihidrite and maghemite. The relative amount of each of them changes with time and with the atmospheric dynamics of each environment.

  14. Modular design of metabolic network for robust production of n-butanol from galactose-glucose mixtures.

    Lim, Hyun Gyu; Lim, Jae Hyung; Jung, Gyoo Yeol

    2015-01-01

    Refactoring microorganisms for efficient production of advanced biofuel such as n-butanol from a mixture of sugars in the cheap feedstock is a prerequisite to achieve economic feasibility in biorefinery. However, production of biofuel from inedible and cheap feedstock is highly challenging due to the slower utilization of biomass-driven sugars, arising from complex assimilation pathway, difficulties in amplification of biosynthetic pathways for heterologous metabolite, and redox imbalance caused by consuming intracellular reducing power to produce quite reduced biofuel. Even with these problems, the microorganisms should show robust production of biofuel to obtain industrial feasibility. Thus, refactoring microorganisms for efficient conversion is highly desirable in biofuel production. In this study, we engineered robust Escherichia coli to accomplish high production of n-butanol from galactose-glucose mixtures via the design of modular pathway, an efficient and systematic way, to reconstruct the entire metabolic pathway with many target genes. Three modular pathways designed using the predictable genetic elements were assembled for efficient galactose utilization, n-butanol production, and redox re-balancing to robustly produce n-butanol from a sugar mixture of galactose and glucose. Specifically, the engineered strain showed dramatically increased n-butanol production (3.3-fold increased to 6.2 g/L after 48-h fermentation) compared to the parental strain (1.9 g/L) in galactose-supplemented medium. Moreover, fermentation with mixtures of galactose and glucose at various ratios from 2:1 to 1:2 confirmed that our engineered strain was able to robustly produce n-butanol regardless of sugar composition with simultaneous utilization of galactose and glucose. Collectively, modular pathway engineering of metabolic network can be an effective approach in strain development for optimal biofuel production with cost-effective fermentable sugars. To the best of our

  15. Metabolic Engineering for Production of Biorenewable Fuels and Chemicals: Contributions of Synthetic Biology

    Jarboe, Laura R.; Zhang, Xueli; Wang, Xuan; Moore, Jonathan C.; Shanmugam, K. T.; Ingram, Lonnie O.

    2010-01-01

    Production of fuels and chemicals through microbial fermentation of plant material is a desirable alternative to petrochemical-based production. Fermentative production of biorenewable fuels and chemicals requires the engineering of biocatalysts that can quickly and efficiently convert sugars to target products at a cost that is competitive with existing petrochemical-based processes. It is also important that biocatalysts be robust to extreme fermentation conditions, biomass-derived inhibito...

  16. Evaluation and identification of histamine-forming bacteria on fish products of middle Adriatic Sea

    Rocco Mancusi

    2013-04-01

    Full Text Available Regulation EU 2073/2005 sets maximum concentration for histamine in fish and products thereof. To meet these criteria, manufacturers have to define performance objectives, such as the maximum allowed prevalence and number/activity of histamine-producing bacteria at relevant stage of production. In order to assess the presence and decarboxylase activity of contaminant bacteria we examined 51 samples of blue fish caught and processed in Emilia Romagna region. We collected 50 gr of fish (skin and gills or the entire product from 10 sample units from every lot. Analytical samples were cultured in Trypticase Soy Broth supplemented with histidine and pyridoxal HCl. Histamine was measured with an electrochemical biosensor after incubation at both 37°C for 24 h and 18-22°C for 48 h. Enrichments that showed relevant enzymatic activity were seeded on Niven agar to isolate suspected colonies and DNA extracts from these bacteria were analyzed by polymerase chain reaction (PCR for detecting specific sequences of the gene encoding pyridoxaldependent histidine decarboxylase (HDC. Overall, 29.4% samples showed relevant production of histamine in broth cultures (above a cut-off value set at 250 ng/mL and 53.3% of them (8 out of 15 samples allowed detection of HDC positive strains. All of them were typed as Morganella, which appears to be the most common of fish caught in middle Adriatic sea. Ten out of the twelve positive samples with enrichment cultures incubated at both 37 and 18-22°C (83% showed higher decarboxylase activity at room temperature, suggesting the presence of psychrotolerant strains. In addition, the prevalence of histamine-producing bacteria was higher at retail than at production level, probably as a consequence of manipulations and cross-contamination. The risk correlated to development of histamine-producing psychrotolerans bacteria cannot be controlled only with storage temperature: it is necessary for the food business operators to

  17. Theoretical Calculations on Sediment Transport on Titan, and the Possible Production of Streamlined Forms

    Burr, D. M.; Emery, J. P.; Lorenz, R. D.

    2005-01-01

    The Cassini Imaging Science System (ISS) has been returning images of Titan, along with other Saturnian satellites. Images taken through the 938 nm methane window see down to Titan's surface. One of the purposes of the Cassini mission is to investigate possible fluid cycling on Titan. Lemniscate features shown recently and radar evidence of surface flow prompted us to consider theoretically the creation by methane fluid flow of streamlined forms on Titan. This follows work by other groups in theoretical consideration of fluid motion on Titan's surface.

  18. An extensive case study of hairy-root cultures for enhanced secondary-metabolite production through metabolic-pathway engineering.

    Mehrotra, Shakti; Rahman, Laiq Ur; Kukreja, Arun Kumar

    2010-08-23

    An intrinsic improvement is taking place in the methodologies for the development of culture systems with first-rate production of plant-based molecules. The blending of HR (hairy root) cultures with ME (metabolic engineering) approaches offers new insights into, and possibilities for, improving the system productivity for known and/or novel high-value plant-derived active compounds. The introduction and expression of foreign genes in plants results in improvement of cellular activities by manipulating enzymatic, regulatory and transport function of the cell. The rational amendments in the rate-limiting steps of a biosynthetic pathway as well as inactivating the inefficient pathway(s) for by-product formation can be accomplished either through single-step engineering or through the multi-step engineering. The hierarchical control of any metabolic process can lead the engineer to apply the ME ideas and principles to any of the strata, including transcriptional, moving on to translational and enzymatic activity. The HR culture systems offer a remarkable potential for commercial production of a number of low-volume, but high-value, secondary metabolites. Taking HR as a model system, in the present review, we discuss engineering principles and perceptions to exploit secondary-metabolite pathways for the production of important bioactive compounds. We also talk about requisites and possible challenges that occur during ME, with emphasis on examples of various HR systems. Furthermore, it also highlights the utilization of global information obtained from '-omic' platforms in order to explore pathway architecture, structural and functional aspects of important enzymes and genes that can support the design of sets of engineering, resulting in the generation of wide-ranging views of DNA sequence-to-metabolite passageway networking and their control to obtain desired results.

  19. Metabolic Design of Corynebacterium glutamicum for Production of l-Cysteine with Consideration of Sulfur-Supplemented Animal Feed.

    Joo, Young-Chul; Hyeon, Jeong Eun; Han, Sung Ok

    2017-06-14

    l-Cysteine is a valuable sulfur-containing amino acid widely used as a nutrition supplement in industrial food production, agriculture, and animal feed. However, this amino acid is mostly produced by acid hydrolysis and extraction from human or animal hairs. In this study, we constructed recombinant Corynebacterium glutamicum strains that overexpress combinatorial genes for l-cysteine production. The aims of this work were to investigate the effect of the combined overexpression of serine acetyltransferase (CysE), O-acetylserine sulfhydrylase (CysK), and the transcriptional regulator CysR on l-cysteine production. The CysR-overexpressing strain accumulated approximately 2.7-fold more intracellular sulfide than the control strain (empty pMT-tac vector). Moreover, in the resulting CysEKR recombinant strain, combinatorial overexpression of genes involved in l-cysteine production successfully enhanced its production by approximately 3.0-fold relative to that in the control strain. This study demonstrates a biotechnological model for the production of animal feed supplements such as l-cysteine using metabolically engineered C. glutamicum.

  20. Secondary Waste Form Screening Test Results—THOR® Fluidized Bed Steam Reforming Product in a Geopolymer Matrix

    Pires, Richard P.; Westsik, Joseph H.; Serne, R. Jeffrey; Mattigod, Shas V.; Golovich, Elizabeth C.; Valenta, Michelle M.; Parker, Kent E.

    2011-07-14

    Screening tests are being conducted to evaluate waste forms for immobilizing secondary liquid wastes from the Hanford Tank Waste Treatment and Immobilization Plant (WTP). Plans are underway to add a stabilization treatment unit to the Effluent Treatment Facility to provide the needed capacity for treating these wastes from WTP. The current baseline is to use a Cast Stone cementitious waste form to solidify the wastes. Through a literature survey, DuraLith alkali-aluminosilicate geopolymer, fluidized-bed steam reformation (FBSR) granular product encapsulated in a geopolymer matrix, and a Ceramicrete phosphate-bonded ceramic were identified both as candidate waste forms and alternatives to the baseline. These waste forms have been shown to meet waste disposal acceptance criteria, including compressive strength and universal treatment standards for Resource Conservation and Recovery Act (RCRA) metals (as measured by the toxicity characteristic leaching procedure [TCLP]). Thus, these non-cementitious waste forms should also be acceptable for land disposal. Information is needed on all four waste forms with respect to their capability to minimize the release of technetium. Technetium is a radionuclide predicted to be in the secondary liquid wastes in small quantities, but the Integrated Disposal Facility (IDF) risk assessment analyses show that technetium, even at low mass, produces the largest contribution to the estimated IDF disposal impacts to groundwater.