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Huan Wang, Zhongling Yang, Shuo Du, Lin Ma, Yao Liao, Yujie Wang, Ian Toth, Jiaqin Fan
Identifying phytopathogen proteins that are differentially expressed in the course of establishing an infection is important to better understand the infection process. In vitro approaches, using plant extracts added to culture medium, have been used to identify such proteins but the biological relevance of these findings for in planta infection are often uncertain until confirmed by in vivo studies. Here, we compared proteins of Pectobacterium carotovorum subsp. carotovorum strain PccS1 differentially expressed in LB medium supplemented with extracts of the ornamental plant Zantedeschia elliotiana cultivar 'Black magic' (in vitro) and in plant tissues (in vivo) by two-dimensional electrophoresis coupled with mass spectrometry...
September 27, 2016: Molecular Plant Pathology
Sambit Roy, Tapas Patra, Taniya Golder, Soniya Chatterjee, Hemanta Koley, Ranjan K Nandy
Orthologs search identified that the Vibrio cholerae gluconate (Gnt) utilization system minimally consisted of the Entner-Doudoroff (ED) pathway (edd and eda) and three other genes namely gntU, gntK and gntR This system appeared unique, by genomic organization of component genes into two operons transcribed in opposite directions. In silico analysis indicated GntU as inner-membrane protein functioning for transport and GntK as kinase with cytosolic localization which generates Gnt6P, then metabolized through the ED pathway...
September 19, 2016: Pathogens and Disease
Eui-Jin Kim, Chang-Hao Wu, Michael W W Adams, Y-H Percival Zhang
Hydrogen production by water splitting energized by biomass sugars is one of the most promising technologies for distributed green H2 production. Direct H2 generation from NADPH, catalysed by an NADPH-dependent, soluble [NiFe]-hydrogenase (SH1) is thermodynamically unfavourable, resulting in slow volumetric productivity. We designed the biomimetic electron transport chain from NADPH to H2 by the introduction of an oxygen-insensitive electron mediator benzyl viologen (BV) and an enzyme (NADPH rubredoxin oxidoreductase, NROR), catalysing electron transport between NADPH and BV...
November 2, 2016: Chemistry: a European Journal
Alina Uifălean, Stefanie Schneider, Philipp Gierok, Corina Ionescu, Cristina Adela Iuga, Michael Lalk
Despite substantial research, the understanding of the chemopreventive mechanisms of soy isoflavones remains challenging. Promising tools, such as metabolomics, can provide now a deeper insight into their biochemical mechanisms. The purpose of this study was to offer a comprehensive assessment of the metabolic alterations induced by genistein, daidzein and a soy seed extract on estrogen responsive (MCF-7) and estrogen non-responsive breast cancer cells (MDA-MB-231), using a global metabolomic approach. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that all test compounds induced a biphasic effect on MCF-7 cells and only a dose-dependent inhibitory effect on MDA-MB-231 cells...
2016: International Journal of Molecular Sciences
Rui Huang, Hui Chen, Chao Zhong, Jae Eung Kim, Yi-Heng Percival Zhang
Coenzyme engineering that changes NAD(P) selectivity of redox enzymes is an important tool in metabolic engineering, synthetic biology, and biocatalysis. Here we developed a high throughput screening method to identify mutants of 6-phosphogluconate dehydrogenase (6PGDH) from a thermophilic bacterium Moorella thermoacetica with reversed coenzyme selectivity from NADP(+) to NAD(+). Colonies of a 6PGDH mutant library growing on the agar plates were treated by heat to minimize the background noise, that is, the deactivation of intracellular dehydrogenases, degradation of inherent NAD(P)H, and disruption of cell membrane...
2016: Scientific Reports
Jiao Meng, Baiyun Wang, Dingyu Liu, Tao Chen, Zhiwen Wang, Xueming Zhao
BACKGROUND: Succinate has been identified by the U.S. Department of Energy as one of the top 12 building block chemicals, which can be used as a specialty chemical in the agricultural, food, and pharmaceutical industries. Escherichia coli are now one of the most important succinate producing candidates. However, the stoichiometric maximum succinate yield under anaerobic conditions through the reductive branch of the TCA cycle is restricted by NADH supply in E. coli. RESULTS: In the present work, we report a rational approach to increase succinate yield by regulating NADH supply via pentose phosphate (PP) pathway and enhancing flux towards succinate...
2016: Microbial Cell Factories
S A Strumilo, U Czyzewska, M Siemieniuk, J Strumilo, A Tylicki
We studied the effects of insulin and cAMP on the offspring of female rats after daily treatment with these substances over 4 weeks. In adult offspring from cAMP-treated females, activities of pyruvate kinase and glucose-6-phosphate dehydrogenase decreased in the liver and brain and activities of NADP-dependent malate dehydrogenase and 6-phosphogluconate dehydrogenase decreased in the liver. In the offspring of insulin-treated females, we observed only activation of glucose-6-phosphate dehydrogenase and malate dehydrogenase in the liver and only in females...
July 2016: Bulletin of Experimental Biology and Medicine
Mohammad Mubinur Rahman, Martina Andberg, Anu Koivula, Juha Rouvinen, Nina Hakulinen
L-Arabinonate dehydratase (EC and D-xylonate dehydratase (EC are two enzymes that are involved in a nonphosphorylative oxidation pathway of pentose sugars. L-Arabinonate dehydratase converts L-arabinonate into 2-dehydro-3-deoxy-L-arabinonate, and D-xylonate dehydratase catalyzes the dehydration of D-xylonate to 2-dehydro-3-deoxy-D-xylonate. L-Arabinonate and D-xylonate dehydratases belong to the IlvD/EDD family, together with 6-phosphogluconate dehydratases and dihydroxyacid dehydratases...
August 2016: Acta Crystallographica. Section F, Structural Biology Communications
Yingna Qiu, Ning Cong, Meng Liang, Yongqiang Wang, Jianshe Wang
In this paper, we investigated the multi-target effect of myricetin as a therapeutic for cardiovascular disease, using an acute ischemia/reperfusion-induced myocardial injury model to gain insight into its mechanism of action. The compound-target interaction profiles of myricetin were determined using a combination of text mining, chemometric and chemogenomic methods. The effect of myricetin on cardiac function was investigated by carrying out experiments in rats subjected to ischemia/reperfusion (I/R) using Langendorff retrograde perfusion technology...
August 2, 2016: Cardiovascular Toxicology
Lili Guo, Andrew J Worth, Clementina Mesaros, Nathaniel W Snyder, Jerry D Glickson, Ian A Blair
RATIONALE: Mass spectrometric (MS) analysis of low molecular weight polar metabolites can be challenging because of poor chromatographic resolution of isomers and insufficient ionization efficiency. These metabolites include intermediates in key metabolic pathways, such as glycolysis, the pentose phosphate pathway, and the Krebs cycle. Therefore, sensitive, specific, and comprehensive quantitative analysis of these metabolites in biological fluids or cell culture models can provide insight into multiple disease states where perturbed metabolism plays a role...
August 30, 2016: Rapid Communications in Mass Spectrometry: RCM
Marie-Christin Lutterbey, Antje von Schaewen
We recently described that all three 6-phosphogluconate dehydrogenase (6PGDH) isoforms of Arabidopsis (PGD) with similar length show dual localization: PGD1 and PGD3 in the cytosol and in plastids, and PGD2 in the cytosol and in peroxisomes. We set out to investigate heterodimer formation, however due to only weak homodimerization of all Arabidopsis PGD isoforms in yeast cells, we conducted further protein-protein interaction studies in planta to investigate homomer versus heteromer formation and their sub-cellular localization...
July 1, 2016: Plant Signaling & Behavior
Mehrdad Ghashghaeinia, Daniela Giustarini, Pavla Koralkova, Martin Köberle, Kousi Alzoubi, Rosi Bissinger, Zohreh Hosseinzadeh, Peter Dreischer, Ingolf Bernhardt, Florian Lang, Mahmoud Toulany, Thomas Wieder, Renata Mojzikova, Ranieri Rossi, Ulrich Mrowietz
In mature erythrocytes, glucose-6-phosphate dehydrogenase (G6PDH) and 6-phosphogluconate dehydrogenase (6PGDH) yield NADPH, a crucial cofactor of the enzyme glutathione reductase (GR) converting glutathione disulfide (GSSG) into its reduced state (GSH). GSH is essential for detoxification processes in and survival of erythrocytes. We explored whether the anti-inflammatory compounds Bay 11-7082, parthenolide and dimethyl fumarate (DMF) were able to completely deplete a common target (GSH), and to impair the function of upstream enzymes of GSH recycling and replenishment...
2016: Scientific Reports
Carolin Bisterfeld, Thomas Classen, Irene Küberl, Birgit Henßen, Alexander Metz, Holger Gohlke, Jörg Pietruszka
The 2-deoxy-d-ribose-5-phosphate aldolase (DERA) offers access to highly desirable building blocks for organic synthesis by catalyzing a stereoselective C-C bond formation between acetaldehyde and certain electrophilic aldehydes. DERA´s potential is particularly highlighted by the ability to catalyze sequential, highly enantioselective aldol reactions. However, its synthetic use is limited by the absence of an enantiocomplementary enzyme. Here, we introduce the concept of homologous grafting to identify stereoselectivity-determining amino acid positions in DERA...
2016: PloS One
Jan-Moritz Sutter, Julia-Beate Tästensen, Ulrike Johnsen, Jörg Soppa, Peter Schönheit
UNLABELLED: The halophilic archaeon Haloferax volcanii has been proposed to degrade glucose via the semiphosphorylative Entner-Doudoroff (spED) pathway. So far, the key enzymes of this pathway, glucose dehydrogenase (GDH), gluconate dehydratase (GAD), and 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase (KDPGA), have not been characterized, and their functional involvement in glucose degradation has not been demonstrated. Here we report that the genes HVO_1083 and HVO_0950 encode GDH and KDPGA, respectively...
August 15, 2016: Journal of Bacteriology
S Elf, R Lin, S Xia, Y Pan, C Shan, S Wu, S Lonial, M Gaddh, M L Arellano, H J Khoury, F R Khuri, B H Lee, T J Boggon, J Fan, J Chen
The oxidative pentose phosphate pathway (PPP) is crucial for cancer cell metabolism and tumor growth. We recently reported that targeting a key oxidative PPP enzyme, 6-phosphogluconate dehydrogenase (6PGD), using our novel small-molecule 6PGD inhibitors Physcion and its derivative S3, shows anticancer effects. Notably, humans with genetic deficiency of either 6PGD or another oxidative PPP enzyme, glucose-6-phosphate dehydrogenase, exhibit non-immune hemolytic anemia upon exposure to aspirin and various antimalarial drugs...
June 6, 2016: Oncogene
Ying Li, Hua Cong, Bingnan Liu, Jinzhu Song, Xueying Sun, Junzheng Zhang, Qian Yang
Relieving the feedback inhibition of key enzymes in a metabolic pathway is frequently the first step of producer-strain construction by genetic engineering. However, the strict feedback regulation exercised by microorganisms in methionine biosynthesis often makes it difficult to produce methionine at a high level. In this study, Corynebacterium glutamicum ATCC 13032 was metabolically engineered for methionine production. First, the metD gene encoding the methionine uptake system was deleted to achieve extracellular accumulation of methionine...
September 2016: Antonie Van Leeuwenhoek
Endang Purwantini, Lacy Daniels, Biswarup Mukhopadhyay
UNLABELLED: Phthiocerol dimycocerosates (PDIM) are a group of cell surface-associated apolar lipids of Mycobacterium tuberculosis and closely related mycobacteria, such as Mycobacterium bovis and Mycobacterium leprae A characteristic methoxy group of these lipids is generated from the methylation of a hydroxyl group of the direct precursors, the phthiotriols. The precursors arise from the reduction of phthiodiolones, the keto intermediates, by a ketoreductase. The putative phthiodiolone ketoreductase (PKR) is encoded by Rv2951c in M...
August 1, 2016: Journal of Bacteriology
Ioannis Papapetridis, Marlous van Dijk, Arthur P A Dobbe, Benjamin Metz, Jack T Pronk, Antonius J A van Maris
BACKGROUND: Acetic acid, an inhibitor of sugar fermentation by yeast, is invariably present in lignocellulosic hydrolysates which are used or considered as feedstocks for yeast-based bioethanol production. Saccharomyces cerevisiae strains have been constructed, in which anaerobic reduction of acetic acid to ethanol replaces glycerol formation as a mechanism for reoxidizing NADH formed in biosynthesis. An increase in the amount of acetate that can be reduced to ethanol should further decrease acetic acid concentrations and enable higher ethanol yields in industrial processes based on lignocellulosic feedstocks...
2016: Microbial Cell Factories
C Luna, E Serrano, J Domingo, A Casao, R Pérez-Pé, J A Cebrián-Pérez, T Muiño-Blanco
Spermatozoa require substantially more ATP than other cells, not only for sustaining sperm motility but also for regulating protein phosphorylation during capacitation. In this study, we have reported for the first time the presence of the two key enzymes of the pentose phosphate pathway (PPP), glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in ovine spermatozoa by indirect immunofluorescence, Western blotting, in-gel activity, and reverse transcription polymerase chain reaction analysis...
August 2016: Theriogenology
Álvaro D Fernández-Fernández, Francisco J Corpas
NADPH, whose regeneration is critical for reductive biosynthesis and detoxification pathways, is an essential component in cell redox homeostasis. Peroxisomes are subcellular organelles with a complex biochemical machinery involved in signaling and stress processes by molecules such as hydrogen peroxide (H2O2) and nitric oxide (NO). NADPH is required by several peroxisomal enzymes involved in β-oxidation, NO, and glutathione (GSH) generation. Plants have various NADPH-generating dehydrogenases, one of which is 6-phosphogluconate dehydrogenase (6PGDH)...
2016: Scientifica
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