Three-Dimensional Graphene Matrix-Supported and Thylakoid Membrane-Based High-Performance Bioelectrochemical Solar Cell

DEFF Research Database (Denmark)

Pankratova, Galina; Pankratov, Dmitrii; Di Bari, Chiara

2018-01-01

A combination of thylakoid membranes (TMs) as photobiocatalysts with high-surface-area electroactive materials could hold great potential for sustainable “green” solar energy conversion. We have studied the orientated immobilization of TMs on high-surface-area graphene electrodes, which were...

Structure and dynamics of thylakoids in land plants

DEFF Research Database (Denmark)

Pribil, Mathias; Labs, Mathias; Leister, Dario

2014-01-01

Thylakoids of land plants have a bipartite structure, consisting of cylindrical grana stacks, made of membranous discs piled one on top of the other, and stroma lamellae which are helically wound around the cylinders. Protein complexes predominantly located in the stroma lamellae and grana end....... Depending on light conditions, thylakoid membranes undergo dynamic structural changes that involve alterations in granum diameter and height, vertical unstacking of grana, and swelling of the thylakoid lumen. This plasticity is realized predominantly by reorganization of the supramolecular structure...

Photosystem II solubilizes as a monomer by mild detergent treatment of unstacked thylakoid membranes

NARCIS (Netherlands)

Dekker, Jan P.; Germano, Marta; Roon, Henny van; Boekema, Egbert J.

2002-01-01

We studied the aggregation state of Photosystem II in stacked and unstacked thylakoid membranes from spinach after a quick andmild solubilization with the non-ionic detergent n-dodecyl-α,D-maltoside, followed by analysis by diode-array-assisted gel filtration chromatography and electron microscopy.

The Zinc-Finger Thylakoid-Membrane Protein FIP Is Involved With Abiotic Stress Response in Arabidopsis thaliana

Directory of Open Access Journals (Sweden)

Karina L. Lopes

2018-04-01

Full Text Available Many plant genes have their expression modulated by stress conditions. Here, we used Arabidopsis FtsH5 protease, which expression is regulated by light stress, as bait in a yeast two-hybrid screen to search for new proteins involved in the stress response. As a result, we found FIP (FtsH5 Interacting Protein, which possesses an amino proximal cleavable transit peptide, a hydrophobic membrane-anchoring region, and a carboxyl proximal C4-type zinc-finger domain. In vivo experiments using FIP fused to green fluorescent protein (GFP showed a plastid localization. This finding was corroborated by chloroplast import assays that showed FIP inserted in the thylakoid membrane. FIP expression was down-regulated in plants exposed to high light intensity, oxidative, salt, and osmotic stresses, whereas mutant plants expressing low levels of FIP were more tolerant to these abiotic stresses. Our data shows a new thylakoid-membrane protein involved with abiotic stress response in Arabidopsis thaliana.

Nonlinear Dielectric Spectroscopy as an Indirect Probe of Metabolic Activity in Thylakoid Membrane

Directory of Open Access Journals (Sweden)

John H. Miller

2011-01-01

Full Text Available Nonlinear dielectric spectroscopy (NDS is a non-invasive probe of cellular metabolic activity with potential application in the development of whole-cell biosensors. However, the mechanism of NDS interaction with metabolic membrane proteins is poorly understood, partly due to the inherent complexity of single cell organisms. Here we use the light-activated electron transport chain of spinach thylakoid membrane as a model system to study how NDS interacts with metabolic activity. We find protein modification, as opposed to membrane pump activity, to be the dominant source of NDS signal change in this system. Potential mechanisms for such protein modifications include reactive oxygen species generation and light-activated phosphorylation.

The mechanism of anthracene interaction with photosynthetic apparatus: A study using intact cells, thylakoid membranes and PS II complexes isolated from Chlamydomonas reinhardtii

International Nuclear Information System (INIS)

Aksmann, Anna; Shutova, Tatiana; Samuelsson, Goeran; Tukaj, Zbigniew

2011-01-01

Intact cells of Chlamydomonas reinhardtii as well as isolated thylakoid membranes and photosystem II complexes were used to examine a possible mechanism of anthracene (ANT) interaction with the photosynthetic apparatus. Since ANT concentrations above 1 mM were required to significantly inhibit the rate of oxygen evolution in PS II membrane fragments it may indicate that the toxicant did not directly interact with this photosystem. On the other hand, stimulation of oxygen uptake by ANT-treated thylakoids suggested that ANT could either act as an artificial electron acceptor in the photosynthetic electron transport chain or function as an uncoupler. Electron transfer from excited chlorophyll to ANT is impossible due to the very low reduction potential of ANT and therefore we propose that toxic concentrations of ANT increase the thylakoid membrane permeability and thereby function as an uncoupler, enhancing electron transport in vitro. Hence, its unspecific interference with photosynthetic membranes in vitro suggests that the inhibitory effect observed on intact cell photosynthesis is caused by uncoupling of phosphorylation.

Enhanced thermal stability of the thylakoid membranes from spruce. A comparison with selected angiosperms

Czech Academy of Sciences Publication Activity Database

Karlický, Václav; Kurasová, Irena; Ptáčková, B.; Večeřová, Kristýna; Urban, Otmar; Špunda, Vladimír

2016-01-01

Roč. 130, 1-3 (2016), s. 357-371 ISSN 0166-8595 R&D Projects: GA MŠk(CZ) LO1415; GA ČR GA13-28093S Institutional support: RVO:67179843 Keywords : Norway spruce * Thermal stability * Circular dichroism * Photosystem II organization * Thylakoid membrane Subject RIV: ED - Physiology Impact factor: 3.864, year: 2016

Red Bell Pepper Chromoplasts Exhibit in Vitro Import Competency and Membrane Targeting of Passenger Proteins from the Thylakoidal Sec and ΔpH Pathways but Not the Chloroplast Signal Recognition Particle Pathway1

Science.gov (United States)

Summer, Elizabeth J.; Cline, Kenneth

1999-01-01

Chloroplast to chromoplast development involves new synthesis and plastid localization of nuclear-encoded proteins, as well as changes in the organization of internal plastid membrane compartments. We have demonstrated that isolated red bell pepper (Capsicum annuum) chromoplasts contain the 75-kD component of the chloroplast outer envelope translocon (Toc75) and are capable of importing chloroplast precursors in an ATP-dependent fashion, indicating a functional general import apparatus. The isolated chromoplasts were able to further localize the 33- and 17-kD subunits of the photosystem II O2-evolution complex (OE33 and OE17, respectively), lumen-targeted precursors that utilize the thylakoidal Sec and ΔpH pathways, respectively, to the lumen of an internal membrane compartment. Chromoplasts contained the thylakoid Sec component protein, cpSecA, at levels comparable to chloroplasts. Routing of OE17 to the lumen was abolished by ionophores, suggesting that routing is dependent on a transmembrane ΔpH. The chloroplast signal recognition particle pathway precursor major photosystem II light-harvesting chlorophyll a/b protein failed to associate with chromoplast membranes and instead accumulated in the stroma following import. The Pftf (plastid fusion/translocation factor), a chromoplast protein, integrated into the internal membranes of chromoplasts during in vitro assays, and immunoblot analysis indicated that endogenous plastid fusion/translocation factor was also an integral membrane protein of chromoplasts. These data demonstrate that the internal membranes of chromoplasts are functional with respect to protein translocation on the thylakoid Sec and ΔpH pathways. PMID:9952453

Mediator-assisted photocurrent extraction from the thylakoids

International Nuclear Information System (INIS)

Yu, Yue; Zuo, Fulin; Li, Chen-Zhong

2014-01-01

Photocurrent extracted from the thylakoids has been studied as a function of electron mediator concentration. Phenazine methosulfate is used to facilitate the charge transfer from the thylakoid's charge transport chain to the outside medium. The photocurrent has been shown to originate from the photosynthesis on the thylakoid membranes. Comparing with a previous study using para-Benzoquinone as the mediator, a similar peak effect in the photocurrent as a function of concentration is observed, but the magnitude of the current is nearly a thousand times greater. A semi-quantitative analysis is presented to explain the data found in those systems

Overaccumulation of glycine betaine makes the function of the thylakoid membrane better in wheat under salt stress

Directory of Open Access Journals (Sweden)

Fengxia Tian

2017-02-01

Full Text Available Wheat (Triticum aestivum L. lines T1, T4, and T6 were genetically modified to increase glycine betaine (GB synthesis by introduction of the BADH (betaine aldehyde dehydrogenase, BADH gene from mountain spinach (Atriplex hortensis L.. These transgenic lines and WT of wheat (T. aestivum L. were used to study the effect of increased GB synthesis on wheat tolerance to salt stress. Salt stress due to 200 mmol L−1 NaCl impaired the photosynthesis of the four wheat lines, as indicated by declines in net photosynthetic rate (Pn, stomatal conductance (Gs, maximum photochemical efficiency of PSII (Fv/Fm, and actual photochemical efficiency of PSII (ФPSII and an increase in intercellular CO2 concentration (Ci. In comparison with WT, the effect of salinity on the three transgenic lines was mild. Salt stress caused disadvantageous changes in lipids and their fatty acid compositions in the thylakoid membrane of the transgenic lines and WT. Under salt stress, the three transgenic lines showed slightly higher chlorophyll and carotenoid contents and higher Hill reaction activities and Ca2+-ATPase activity than WT. All the results suggest that overaccumulation of GB resulting from introduction of the BADH gene can enhance the salt tolerance of transgenic plants, especially in the protection of the components and function of thylakoid membranes, thereby making photosynthesis better. Changes in lipids and fatty acid compositions in the thylakoid membrane may be involved in the increased salt stress tolerance of the transgenic lines.

The chloroplast thylakoid membrane system is a molecular conveyor belt.

Science.gov (United States)

Critchley, C

1988-10-01

Light drives photosynthesis, but paradoxically light is also the most variable environmental factor influencing photosynthesis both qualitatively and quantitatively. The photosynthetic apparatus of higher plants is adaptable in the extreme, as exemplified by its capacity for acclimation to very bright sunny or deeply shaded conditions. It can also respond to rapid changes in light such as sunflecks. In this paper I offer a model that i) explains the thylakoid membrane organisation into grana stacks and stroma lamellae, ii) proposes a role for rapid D1 protein turnover and LHCII phosphorylation, and iii) suggests a mechanism for photoinhibition. I argue that the photosynthetic membrane system is dynamic in three dimensions, so much so that, in the light, it is in constant motion and operates in a manner somewhat analogous to a conveyor belt. D1 protein degradation is proposed to be the motor that drives this system. Photoinhibition is suggested to be due to the arrest of D1 protein turnover.

Two roles of thylakoid lipids in modifying the activity of herbicides which inhibit photosystem II

International Nuclear Information System (INIS)

Kupatt, C.C. Jr.

1985-01-01

Thylakoid lipids may modify the activity of herbicides which inhibit electron transport at the Q/sub B/ protein of photosystem II in two ways: (1) lipids can act as a hydrophobic barrier to a binding site localized close to the loculus of the membrane, and (2) changes in lipid composition can reduce the ability of inhibitors to block electron transport, possibly due to a change in the conformation of the Q/sub B/ protein. The herbicide binding site was localized close to the locular side of the thylakoid membrane by determining the activity of a number of substituted phenylurea and s-triazine herbicides in inverted and non-inverted thylakoids. Quantitative structure-activity relationship analysis showed that inversion of thylakoids reduced the requirement of molecular lipophilicity deemed necessary for phenylurea activity in non-inverted membranes, whereas s-triazines exhibited no differences in the lipophilicity requirement in thylakoid membranes of either orientation. The binding affinity of 14 C-diuron was reduced in bicarbonate-depleted thylakoids relative to reconstituted or control membranes, as is the case with atrazine binding. These observations support a model of the herbicide binding site containing both common and herbicide family specific binding domains. Thylakoids isolated either from detached lambs quarters (Chenopodium album L.) leaves, treated with SAN 6706, or from soybean (Glycine max L.), with norflurazon or pyrazon applied preemergence, exhibited decreased susceptibility to atrazine. The ability of lipid-modifying treatments to decrease the atrazine susceptibility of field-grown soybeans was also investigated

Is There Excitation Energy Transfer between Different Layers of Stacked Photosystem-II-Containing Thylakoid Membranes?

Science.gov (United States)

Farooq, Shazia; Chmeliov, Jevgenij; Trinkunas, Gediminas; Valkunas, Leonas; van Amerongen, Herbert

2016-04-07

We have compared picosecond fluorescence decay kinetics for stacked and unstacked photosystem II membranes in order to evaluate the efficiency of excitation energy transfer between the neighboring layers. The measured kinetics were analyzed in terms of a recently developed fluctuating antenna model that provides information about the dimensionality of the studied system. Independently of the stacking state, all preparations exhibited virtually the same value of the apparent dimensionality, d = 1.6. Thus, we conclude that membrane stacking does not affect the efficiency of the delivery of excitation energy toward the reaction centers but ensures a more compact organization of the thylakoid membranes within the chloroplast and separation of photosystems I and II.

The evolutionarily conserved protein PHOTOSYNTHESIS AFFECTED MUTANT71 is required for efficient manganese uptake at the thylakoid membrane in Arabidopsis

DEFF Research Database (Denmark)

Schneider, Anja; Steinberger, Iris; Herdean, Andrei

2016-01-01

In plants, algae, and cyanobacteria, photosystem II (PSII) catalyzes the light-driven oxidation of water. The oxygen-evolving complex of PSII is a Mn4CaO5 cluster embedded in a well-defined protein environment in the thylakoid membrane. However, transport of manganese and calcium into the thylako...... was restored by supplementation with Mn2+, but not Ca2+. Furthermore, PAM71 suppressed the Mn2+-sensitive phenotype of the yeast mutant Δpmr1. Therefore, PAM71 presumably functions in Mn2+ uptake into thylakoids to ensure optimal PSII performance....

Molecular analysis of a thylakoid K+channel

International Nuclear Information System (INIS)

1999-01-01

The work undertaken sought to use a novel probe to identify and clone plant ion (K) channels. It was also proposed that in vitro biochemical studies of cation transport across purified preparations of thylakoid membrane be employed to characterize a putative K channel in this membrane system. Over the last several years, an enormous data base of partially-sequenced mRNAs and numerous genomes (including those of plants) has evolved and provides a powerful alternative to this brute-force approach to identify and clone cDNAs encoding physiologically important membrane proteins such as channels. The utility of searching genetic databases for relevant sequences, in addition to the difficulty of working with membrane proteins, led to changes in research focus during the granting period. During the course of the funding period, work was finished up which documented the presence of a K channel in the thylakoid membrane and demonstrated that K fluxes through this channel were required for optimal photosynthetic activity, likely due to the requirement for charge balancing of proton flux

Molecular analysis of a thylakoid K+channel

Energy Technology Data Exchange (ETDEWEB)

NONE

1999-09-10

The work undertaken sought to use a novel probe to identify and clone plant ion (K) channels. It was also proposed that in vitro biochemical studies of cation transport across purified preparations of thylakoid membrane be employed to characterize a putative K channel in this membrane system. Over the last several years, an enormous data base of partially-sequenced mRNAs and numerous genomes (including those of plants) has evolved and provides a powerful alternative to this brute-force approach to identify and clone cDNAs encoding physiologically important membrane proteins such as channels. The utility of searching genetic databases for relevant sequences, in addition to the difficulty of working with membrane proteins, led to changes in research focus during the granting period. During the course of the funding period, work was finished up which documented the presence of a K channel in the thylakoid membrane and demonstrated that K fluxes through this channel were required for optimal photosynthetic activity, likely due to the requirement for charge balancing of proton flux.

Ribosomal protein S6 kinase1 coordinates with TOR-Raptor2 to regulate thylakoid membrane biosynthesis in rice.

Science.gov (United States)

Sun, Linxiao; Yu, Yonghua; Hu, Weiqin; Min, Qiming; Kang, Huiling; Li, Yilu; Hong, Yue; Wang, Xuemin; Hong, Yueyun

2016-07-01

Ribosomal protein S6 kinase (S6K) functions as a key component in the target of rapamycin (TOR) pathway involved in multiple processes in eukaryotes. The role and regulation of TOR-S6K in lipid metabolism remained unknown in plants. Here we provide genetic and pharmacological evidence that TOR-Raptor2-S6K1 is important for thylakoid galactolipid biosynthesis and thylakoid grana modeling in rice (Oryza sativa L.). Genetic suppression of S6K1 caused pale yellow-green leaves, defective thylakoid grana architecture. S6K1 directly interacts with Raptor2, a core component in TOR signaling, and S6K1 activity is regulated by Raptor2 and TOR. Plants with suppressed Raptor2 expression or reduced TOR activity by inhibitors mimicked the S6K1-deficient phenotype. A significant reduction in galactolipid content was found in the s6k1, raptor2 mutant or TOR-inhibited plants, which was accompanied by decreased transcript levels of the set of genes such as lipid phosphate phosphatase α5 (LPPα5), MGDG synthase 1 (MGD1), and DGDG synthase 1 (DGD1) involved in galactolipid synthesis, compared to the control plants. Moreover, loss of LPPα5 exhibited a similar phenotype with pale yellow-green leaves. These results suggest that TOR-Raptor2-S6K1 is important for modulating thylakoid membrane lipid biosynthesis, homeostasis, thus enhancing thylakoid grana architecture and normal photosynthesis ability in rice. Copyright © 2016 Elsevier B.V. All rights reserved.

Operation of trans-thylakoid thiol-metabolizing pathways in photosynthesis

Directory of Open Access Journals (Sweden)

Mohamed eKaramoko

2013-11-01

Full Text Available Thiol oxidation to disulfides and the reverse reaction, i.e. disulfide reduction to free thiols, are under the control of catalysts in vivo. Enzymatically assisted thiol-disulfide chemistry is required for the biogenesis of all energy-transducing membrane systems. However, until recently, this had only been demonstrated for the bacterial plasma membrane. Long considered to be vacant, the thylakoid lumen has now moved to the forefront of photosynthesis research with the realization that its proteome is far more complicated than initially anticipated. Several lumenal proteins are known to be disulfide bonded in Arabidopsis, highlighting the importance of sulfhydryl oxidation in the thylakoid lumen. While disulfide reduction in the plastid stroma is known to activate several enzymatic activities, it appears that it is the reverse reaction, i.e. thiol oxidation that is required for the activity of several lumen-resident proteins. This paradigm for redox regulation in the thylakoid lumen has opened a new frontier for research in the field of photosynthesis. Of particular significance in this context is the discovery of trans-thylakoid redox pathways controlling disulfide bond formation and reduction, which are required for photosynthesis.

Kinetic and spectroscopic studies of cytochrome b-563 in isolated cytochrome b/f complex and in thylakoid membranes

Energy Technology Data Exchange (ETDEWEB)

Hind, G.; Clark, R.D.; Houchins, J.P.

1983-01-01

Extensive studies, performed principally by Hauska, Hurt and collaborators, have shown that a cytochrome (cyt) b/f complex isolated from photosynthetic membranes of spinach or Anabaena catalyzes electron transport from plastoquinol (PQH/sub 2/) to plastocyanin or algal cyt c-552. The complex from spinach thylakoids generated a membrane potential when reconstituted into liposomes, and although the electrogenic mechanism remains unknown, a key role for cyt b-563 is widely accepted. Electrogenesis by a Q-cycle mechanism requires a plastoquinone (PQ) reductase to be associated with the stromal side of the thylakoid b/f complex though this activity has yet to be demonstrated. It seemed possible that more gentle isolation of the complex might yield a form containing additional polypeptides, perhaps including a PQ reductase or a component involved in returning electrons from reduced ferredoxin to the complex in cyclic electron flow. Optimization of the isolation of cyt b/f complex for Hybrid 424 spinach from a growth room was also required. The procedure we devised is compared to the protocol of Hurt and Hauska (1982). 13 references.

Small-angle neutron scattering study of the ultrastructure of chloroplast thylakoid membranes - Periodicity and structural flexibility of the stroma lamellae

DEFF Research Database (Denmark)

Posselt, Dorthe; Nagy, Gergely; Kirkensgaard, Jacob J. K.

2012-01-01

The multilamellar organization of freshly isolated spinach and pea chloroplast thylakoid membranes was studied using small-angle neutron scattering. A broad peak at similar to 0.02 angstrom(-1) is ascribed to diffraction from domains of ordered, unappressed stroma lamellae, revealing a repeat dis...

Characteristics and functionality of appetite-reducing thylakoid powders produced by three different drying processes.

Science.gov (United States)

Östbring, Karolina; Sjöholm, Ingegerd; Sörenson, Henrietta; Ekholm, Andrej; Erlanson-Albertsson, Charlotte; Rayner, Marilyn

2018-03-01

Thylakoids, a chloroplast membrane extracted from green leaves, are a promising functional ingredient with appetite-reducing properties via their lipase-inhibiting effect. Thylakoids in powder form have been evaluated in animal and human models, but no comprehensive study has been conducted on powder characteristics. The aim was to investigate the effects of different isolation methods and drying techniques (drum-drying, spray-drying, freeze-drying) on thylakoids' physicochemical and functional properties. Freeze-drying yielded thylakoid powders with the highest lipase-inhibiting capacity. We hypothesize that the specific macromolecular structures involved in lipase inhibition were degraded to different degrees by exposure to heat during spray-drying and drum-drying. We identified lightness (Hunter's L-value), greenness (Hunter's a-value), chlorophyll content and emulsifying capacity to be correlated to lipase-inhibiting capacity. Thus, to optimize the thylakoids functional properties, the internal membrane structure indicated by retained green colour should be preserved. This opens possibilities to use chlorophyll content as a marker for thylakoid functionality in screening processes during process optimization. Thylakoids are heat sensitive, and a mild drying technique should be used in industrial production. Strong links between physicochemical parameters and lipase inhibition capacity were found that can be used to predict functionality. The approach from this study can be applied towards production of standardized high-quality functional food ingredients. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

The transporter SynPAM71 is located in the plasma membrane and thylakoids, and mediates manganese tolerance in Synechocystis PCC6803

DEFF Research Database (Denmark)

Gandini, Chiara; Schmidt, Sidsel Birkelund; Husted, Søren

2017-01-01

symptoms were observed in WT cells exposed to excess Mn. Moreover, CyanoP, which is involved in the early steps of PSII assembly, is massively upregulated in ΔSynPAM71. SynPAM71 was detected in both the plasma membrane and, to a lesser extent, the thylakoid membranes. Our results suggest that SynPAM71......Manganese (Mn) is an essential constituent of photosystem II (PSII) and therefore indispensable for oxygenic photosynthesis. Very little is known about how Mn is transported, delivered and retained in photosynthetic cells. Recently, the thylakoid-localized transporter PAM71 has been linked...... to chloroplast Mn homeostasis in Arabidopsis thaliana. Here, we characterize the function of its homolog in Synechocystis (SynPAM71). We used a loss-of-function line (ΔSynPAM71), wild-type (WT) cells exposed to Mn stress and strains expressing a tagged variant of SynPAM71 to characterize the role of SynPAM71...

Towards an understanding of wheat chloroplasts: a methodical investigation of thylakoid proteome.

Science.gov (United States)

Kamal, Abu Hena Mostafa; Cho, Kun; Komatsu, Setsuko; Uozumi, Nobuyuki; Choi, Jong-Soon; Woo, Sun Hee

2012-05-01

We utilized Percoll density gradient centrifugation to isolate and fractionate chloroplasts of Korean winter wheat cultivar cv. Kumgang (Triticum aestivum L.). The resulting protein fractions were separated by one dimensional polyacrylamide gel electrophoresis (1D-PAGE) coupled with LTQ-FTICR mass spectrometry. This enabled us to detect and identify 767 unique proteins. Our findings represent the most comprehensive exploration of a proteome to date. Based on annotation information from the UniProtKB/Swiss-Prot database and our analyses via WoLF PSORT and PSORT, these proteins are localized in the chloroplast (607 proteins), chloroplast stroma (145), thylakoid membrane (342), lumens (163), and integral membranes (166). In all, 67% were confirmed as chloroplast thylakoid proteins. Although nearly complete protein coverage (89% proteins) has been accomplished for the key chloroplast pathways in wheat, such as for photosynthesis, many other proteins are involved in regulating carbon metabolism. The identified proteins were assigned to 103 functional categories according to a classification system developed by the iProClass database and provided through Protein Information Resources. Those functions include electron transport, energy, cellular organization and biogenesis, transport, stress responses, and other metabolic processes. Whereas most of these proteins are associated with known complexes and metabolic pathways, about 13% of the proteins have unknown functions. The chloroplast proteome contains many proteins that are localized to the thylakoids but as yet have no known function. We propose that some of these familiar proteins participate in the photosynthetic pathway. Thus, our new and comprehensive protein profile may provide clues for better understanding that photosynthetic process in wheat.

Purification and characterization of a thylakoid protein kinase

International Nuclear Information System (INIS)

Coughlan, S.J.; Hind, G.

1986-01-01

Control of state transitions in the thylakoid by reversible phosphorylation of the light-harvesting chlorophyll a/b protein complex of photosystem II (LHC-II) is modulated by a kinase. The kinase catalyzing this phosphorylation is associated with the thylakoid membrane, and is regulated by the redox state of the plastoquinone pool. The isolation and partial purification from spinach thylakoids of two protein kinases (CPK1, CPK2) of apparent molecular masses 25 kDa and 38 kDa has been reported. Neither enzyme utilizes isolated LHC-II as a substrate. The partial purification of a third protein kinase (LHCK) which can utilize both lysine-rich histones (IIIs and Vs) and isolated LHC-II as substrate has now been purified to homogeneity and characterized by SDS-polyacrylamide gel electrophoresis as a 64 kDa peptide. From a comparison of the two isolation procedures we have concluded that CPK1 is indeed a protein kinase, but has a lower specific activity than that of LHCK. 8 refs., 4 figs

Thylakoids promote release of the satiety hormone cholecystokinin while reducing insulin in healthy humans

DEFF Research Database (Denmark)

Köhnke, Rickard; Lindbo, Agnes; Larsson, Therese

2009-01-01

(CCK, leptin and ghrelin), insulin and blood metabolites (glucose and free fatty acids). RESULTS: The CCK level increased, in particular between the 120 min time-point and onwards, the ghrelin level was reduced at 120 min and leptin level increased at 360 min after intake of the thylakoid-enriched meal....... The insulin level was reduced, whereas glucose concentrations were unchanged. Free fatty acids were reduced between time-point 120 min and onwards after the thylakoid meal. CONCLUSIONS: The addition of thylakoids to energy-dense food promotes satiety signals and reduces insulin response during a single meal......OBJECTIVE: The effects of a promising new appetite suppressor named "thylakoids" (membrane proteins derived from spinach leaves) were examined in a single meal in man. Thylakoids inhibit the lipase/colipase hydrolysis of triacylglycerols in vitro and suppress food intake, decrease body-weight gain...

Thylakoids suppress appetite by increasing cholecystokinin resulting in lower food intake and body weight in high-fat fed mice

DEFF Research Database (Denmark)

Köhnke, Rickard; Lindqvist, Andreas; Göransson, Nathanael

2009-01-01

Thylakoids are membranes isolated from plant chloroplasts which have previously been shown to inhibit pancreatic lipase/colipase catalysed hydrolysis of fat in vitro and induce short-term satiety in vivo. The purpose of the present study was to examine if dietary supplementation of thylakoids cou...

Resonance Raman spectroscopy of xanthophylls in pigment mutant thylakoid membranes of pea.

Science.gov (United States)

Andreeva, Atanaska; Stoitchkova, Katerina; Busheva, Mira; Apostolova, Emilia; Várkonyi, Zsuzsanna; Garab, Gyözö

Low-temperature resonance Raman spectroscopy was used to study the changes in the molecular structure and configuration of the major xanthophylls in thylakoid membranes isolated from mutants of pea with modified pigment content and altered structural organization of their pigment-protein complexes. The Raman spectra contained four known groups of bands, nu(1)-nu(4), which could be assigned to originate mainly from the long wavelength absorbing lutein and neoxanthin upon 514.5 nm and at 488 nm excitations, respectively. The overall configuration of these bound xanthophyll molecules in the mutants appeared to be similar to the wild type, and the configuration in the wild type was almost identical with that in the isolated main chlorophyll a/b light harvesting protein complex of photosystem II (LHCII). Significant differences were found mainly in the region of nu(4) (around 960 cm(-1)), which suggest that the macroorganization of PS II-LHCII supercomplexes and/or of the LHCII-only domains are modified in the mutants compared to the wild type. Copyright 2004 Wiley Periodicals, Inc. Biopolymers, 2004

Analysis of ionic channels by a flash spectrophotometric technique applicable to thylakoid membranes : CF0, the proton channel of the chloroplast ATP synthase, and, for comparison, gramicidin

NARCIS (Netherlands)

Lill, H.; Althoff, Gerd; Junge, Wolfgang

We previously introduced a flash spectrophotometric method to analyze proton conduction by CF0 in vesicles derived from thylakoid membranes (H. Lill, S. Engelbrecht, G. Schönknecht & W. Junge, 1986, Eur. J. Biochem.160:627-634). The unit conductance of CF0, as revealed by this technique, was orders

Tat proteins as novel thylakoid membrane anchors organize a biosynthetic pathway in chloroplasts and increase product yield 5-fold

DEFF Research Database (Denmark)

Henriques de Jesus, Maria Perestrello Ramos; Nielsen, Agnieszka Janina Zygadlo; Mellor, Silas Busck

2017-01-01

to their complex structures. Some of the crucial enzymes catalyzing their biosynthesis are the cytochromes P450 (P450s) situated in the endoplasmic reticulum (ER), powered by electron transfers from NADPH. Dhurrin is a cyanogenic glucoside and its biosynthesis involves a dynamic metabolon formed by two P450s....... Nevertheless, translocation of the pathway from the ER to the chloroplast creates other difficulties, such as the loss of metabolon formation and intermediate diversion into other metabolic pathways. We show here that co-localization of these enzymes in the thylakoid membrane leads to a significant increase...... in product formation, with a concomitant decrease in off-pathway intermediates. This was achieved by exchanging the membrane anchors of the dhurrin pathway enzymes to components of the Twin-arginine translocation pathway, TatB and TatC, which have self-assembly properties. Consequently, we show 5-fold...

Molecular analysis of a thylakoid K+ channel

Energy Technology Data Exchange (ETDEWEB)

NONE

2000-05-01

The work undertaken during the prior granting period sought to use a novel probe to identify and clone plant ion (K) channels. It was also proposed that in vitro biochemical studies of cation transport across purified preparations of thylakoid membrane be employed to characterize a putative K channel in this membrane system. Over the last several years (including those of the previous grant period), an enormous data base of partially-sequenced mRNAs and numerous genomes (including those of plants) has evolved and provides a powerful alternative to this brute-force approach to identify and clone cDNAs ending physiologically important membrane proteins such as channels. The utility of searching genetic databases for relevant sequences, in addition to the difficulty of working with membrane proteins, led to changes in research focus during the prior granting period, and has resulted in the identification of a new class of plant ion channels, which will be the focus of research during the proposed new granting period.

RECOVERY ACT - Thylakoid Assembly and Folded Protein Transport by the Tat Pathway

Energy Technology Data Exchange (ETDEWEB)

Dabney-Smith, Carole [Miami Univ., Oxford, OH (United States)

2016-07-18

Assembly of functional photosystems complete with necessary intrinsic (membrane-bound) and extrinsic proteins requires the function of at least 3 protein transport pathways in thylakoid membranes. Our research focuses on one of those pathways, a unique and essential protein transport pathway found in the chloroplasts of plants, bacteria, and some archaebacteria, the Twin arginine translocation (Tat) system. The chloroplast Tat (cpTat) system is thought to be responsible for the proper location of ~50% of thylakoid lumen proteins, several of which are necessary for proper photosystem assembly, maintenance, and function. Specifically, cpTat systems are unique because they transport fully folded and assembled proteins across ion tight membranes using only three membrane components, Tha4, Hcf106, and cpTatC, and the protonmotive force generated by photosynthesis. Despite the importance of the cpTat system in plants, the mechanism of transport of a folded precursor is not well known. Our long-term goal is to investigate the role protein transport systems have on organelle biogenesis, particularly the assembly of membrane protein complexes in thylakoids of chloroplasts. The objective of this proposal is to correlate structural changes in the membrane-bound cpTat component, Tha4, to the mechanism of translocation of folded-precursor substrates across the membrane bilayer by using a cysteine accessibility and crosslinking approach. Our central hypothesis is that the precursor passes through a proteinaceous pore of assembled Tha4 protomers that have undergone a conformational or topological change in response to transport. This research is predicated upon the observations that Tha4 exists in molar excess in the membrane relative to the other cpTat components; its regulated assembly to the precursor-bound receptor; and our data showing oligomerization of Tha4 into very large complexes in response to transport. Our rationale for these studies is that understanding cp

Supplementation by thylakoids to a high carbohydrate meal decreases feelings of hunger, elevates CCK levels and prevents postprandial hypoglycaemia in overweight women

DEFF Research Database (Denmark)

Stenblom, Eva-Lena; Montelius, Caroline; Östbring, Karolina

2013-01-01

Thylakoids are chlorophyll-containing membranes in chloroplasts that have been isolated from green leaves. It has been previously shown that thylakoids supplemented with a high-fat meal can affect cholecystokinin (CCK), ghrelin, insulin and blood lipids in humans, and can act to suppress food...... intake and prevent body weight gain in rodents. This study investigates the addition of thylakoids to a high carbohydrate meal and its effects upon hunger motivation and fullness, and the levels of glucose, insulin, CCK, ghrelin and tumour necrosis factor (TNF)-alpha in overweight women. Twenty...... moderately overweight female subjects received test meals on three different occasions; two thylakoid enriched and one control, separated by 1 week. The test meals consisted of a high carbohydrate Swedish breakfast, with or without addition of thylakoids. Blood samples and VAS-questionnaires were evaluated...

Quality control of Photosystem II: the mechanisms for avoidance and tolerance of light and heat stresses are closely linked to membrane fluidity of the thylakoids

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Yasusi Yamamoto

2016-08-01

Full Text Available When oxygenic photosynthetic organisms are exposed to excessive light and/or heat, Photosystem II is damaged and electron transport is blocked. In these events, reactive oxygen species, endogenous radicals and lipid peroxidation products generated by photochemical reaction and/or heat cause the damage. Regarding light stress, plants first dissipate excessive light energy captured by light-harvesting chlorophyll protein complexes as heat to avoid the hazards, but once light stress is unavoidable, they tolerate the stress by concentrating damage in a particular protein in photosystem II, i.e. the reaction-center binding D1 protein of Photosystem II. The damaged D1 is removed by specific proteases and replaced with a new copy produced through de novo synthesis (reversible photoinhibition. When light intensity becomes extremely high, irreversible aggregation of D1 occurs and thereby D1 turnover is prevented. Once the aggregated products accumulate in Photosystem II complexes, removal of them by proteases is difficult, and irreversible inhibition of Photosystem II takes place (irreversible photoinhibition. Important is that various aspects of both the reversible and irreversible photoinhibition are highly dependent on the membrane fluidity of the thylakoids. Heat stress-induced inactivation of photosystem II is an irreversible process, which may be also affected by the fluidity of the thylakoid membranes. Here I describe why the membrane fluidity is a key to regulate the avoidance and tolerance of Photosystem II on environmental stresses.

[Effect of high magnesium ion concentration on the electron transport rate and proton exchange in thylakoid membranes in higher plants].

Science.gov (United States)

Ignat'ev, A R; Khorobrykh, S A; Ivanov, B N

2001-01-01

The effects of magnesium ion concentration on the rate of electron transport in isolated pea thylakoids were investigated in the pH range from 4.0 up to 8.0. In the absence of magnesium ions in the medium and in the presence of 5 mM MgCl2 in the experiments not only without added artificial acceptors but also with ferricyanide or methylviologen as an acceptor, this rate had a well-expressed maximum at pH 5.0. It was shown that, after depression to minimal values at pH 5.5-6.5, it gradually rose with increasing pH. An increase in magnesium ion concentration up to 20 mM essentially affected the electron transfer rate: it decreased somewhat at pH 4.0-5.0 but increased at higher pH values. At this magnesium ion concentration, the maximum rate was at pH 6.0-6.5 and the minimum, at pH 7.0. Subsequent rise upon increasing pH to 8.0 was expressed more sharply. The influence of high magnesium ion concentration on the rate of electron transport was not observed in the presence of gramicidin D. It was found that without uncoupler, the changes in the electron transfer rate under the influence of magnesium ions correlated to the changes in the first-order rate constant of the proton efflux from thylakoids. It is supposed that the change in the ability of thylakoids to keep protons by the action of magnesium ions is the result of electrostatic interactions of these ions with the charges on the external surface of membranes. A possible role of regulation of the electron transport rate by magnesium ions in vivo is discussed.

Thylakoid redox signals are integrated into organellar-gene-expression-dependent retrograde signalling in the prors1-1 mutant

Directory of Open Access Journals (Sweden)

Luca eTadini

2012-12-01

Full Text Available Perturbations in organellar gene expression (OGE and the thylakoid redox state (TRS activate retrograde signalling pathways that adaptively modify nuclear gene expression (NGE, according to developmental and metabolic needs. The prors1-1 mutation in Arabidopsis down-regulates the expression of the nuclear gene Prolyl-tRNA Synthetase1 (PRORS1 which acts in both plastids and mitochondria, thereby impairing protein synthesis in both organelles and triggering OGE-dependent retrograde signalling. Because the mutation also affects thylakoid electron transport, TRS-dependent signals may likewise have an impact on the changes in NGE observed in this genotype. In this study, we have investigated whether signals related to TRS are actually integrated into the OGE-dependent retrograde signalling pathway. To this end, the chaos mutation (for chlorophyll a/b binding protein harvesting-organelle specific, which shows a partial loss of PSII antennae proteins and thus a reduction in PSII light absorption capability, was introduced into the prors1-1 mutant background. The resulting double mutant displayed a prors1-1-like reduction in plastid translation rate and a chaos-like decrease in PSII antenna size, whereas the hyper-reduction of the thylakoid electron transport chain, caused by the prors1-1 mutation, was alleviated, as determined by monitoring chlorophyll (Chl fluorescence and thylakoid phosphorylation. Interestingly, a substantial fraction of the nucleus-encoded photosynthesis genes down-regulated in the prors1-1 mutant are expressed at nearly wild-type rates in prors1-1 chaos leaves, and this recovery is reflected in the steady-state levels of their protein products in the chloroplast. We therefore conclude that signals related to photosynthetic electron transport and TRS, and indirectly to carbohydrate metabolism and energy balance, are indeed fed into the OGE-dependent retrograde pathway to modulate NGE and adjust the abundance of chloroplast proteins.

Consumption of thylakoid-rich spinach extract reduces hunger, increases satiety and reduces cravings for palatable food in overweight women

OpenAIRE

Stenblom, Eva-Lena; Egecioglu, Emil; Landin-Olsson, Mona; Erlanson-Albertsson, Charlotte

2015-01-01

Green-plant membranes, thylakoids, have previously been found to increase postprandial release of the satiety hormone GLP-1, implicated in reward signaling. The purpose of this study was to investigate how treatment with a single dose of thylakoids before breakfast affects homeostatic as well as hedonic hunger, measured as wanting and liking for palatable food (VAS). We also examined whether treatment effects were correlated to scores for eating behavior. Compared to placebo, intake of thylak...

Phylogenetic analysis of the thylakoid ATP/ADP carrier reveals new insights into its function restricted to green plants

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Cornelia eSpetea

2012-01-01

Full Text Available ATP is the common energy currency of cellular metabolism in all living organisms. Most of them synthesize ATP in the cytosol or on the mitochondrial inner membrane, whereas land plants, algae and cyanobacteria also produce it on the thylakoid membrane during the light-dependent reactions of photosynthesis. From the site of synthesis, ATP is transported to the site of utilization via intracellular membranes transporters. One major type of ATP transporter is represented by the mitochondrial ADP/ATP carrier family. Here we review a recently characterized member, namely the thylakoid ATP/ADP carrier from Arabidopsis thaliana (AtTAAC. Thus far, no orthologues of this carrier have been characterized in other organisms, although similar sequences can be recognized in many sequenced genomes. Protein Sequence database searches and phylogenetic analyses indicate the absence of TAAC in cyanobacteria and its appearance early in the evolution of photosynthetic eukaryotes. The TAAC clade is composed of carriers found in land plants and some green algae, but no proteins from other photosynthetic taxa, such as red algae, brown algae and diatoms. This implies that TAAC-like sequences arose only once before the divergence of green algae and land plants. Based on these findings, it is proposed that TAAC may have evolved in response to the need of a new activity in higher photosynthetic eukaryotes. This activity may provide the energy to drive reactions during biogenesis and turnover of photosynthetic complexes, which are heterogenously distributed in a thylakoid membrane system composed of appressed and non-appressed regions.

The Arabidopsis thylakoid chloride channel AtCLCe functions in chloride homeostasis and regulation of photosynthetic electron transport

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Andrei eHerdean

2016-02-01

Full Text Available Chloride ions can be translocated across cell membranes through Cl− channels or Cl−/H+ exchangers. The thylakoid-located member of the Cl− channel CLC family in Arabidopsis thaliana (AtCLCe was hypothesized to play a role in photosynthetic regulation based on the initial photosynthetic characterization of clce mutant lines. The reduced nitrate content of Arabidopsis clce mutants suggested a role in regulation of plant nitrate homeostasis. In this study, we aimed to further investigate the role of AtCLCe in the regulation of ion homeostasis and photosynthetic processes in the thylakoid membrane. We report that the size and composition of proton motive force were mildly altered in two independent Arabidopsis clce mutant lines. Most pronounced effects in the clce mutants were observed on the photosynthetic electron transport of dark-adapted plants, based on the altered shape and associated parameters of the polyphasic OJIP kinetics of chlorophyll a fluorescence induction. Other alterations were found in the kinetics of state transition and in the macro-organisation of photosystem II supercomplexes, as indicated by circular dichroism measurements. Pre-treatment with KCl but not with KNO3 restored the wild-type photosynthetic phenotype. Analyses by transmission electron microscopy revealed a bow-like arrangement of the thylakoid network and a large thylakoid-free stromal region in chloroplast sections from the dark-adapted clce plants. Based on these data, we propose that AtCLCe functions in Cl− homeostasis after transition from light to dark, which affects chloroplast ultrastructure and regulation of photosynthetic electron transport.

Photosynthetic electron transport in thylakoid preparations from two marine red algae (Rhodophyta).

Science.gov (United States)

Stewart, A C; Larkum, A W

1983-01-01

Thylakoid membrane preparations active in photosynthetic electron transport have been obtained from two marine red algae, Griffithsia monilis and Anotrichium tenue. High concentrations (0.5-1.0 M) of salts such as phosphate, citrate, succinate and tartrate stabilized functional binding of phycobilisomes to the membrane and also stabilized Photosystem II-catalysed electron-transport activity. High concentrations (1.0 M) of chloride and nitrate, or 30 mM-Tricine/NaOH buffer (pH 7.2) in the absence of salts, detached phycobilisomes and inhibited electron transport through Photosystem II. The O2-evolving system was identified as the electron-transport chain component that was inhibited under these conditions. Washing membranes with buffers containing 1.0-1.5 M-sorbitol and 5-50 mM concentrations of various salts removed the outer part of the phycobilisome but retained 30-70% of the allophycocyanin 'core' of the phycobilisome. These preparations were 30-70% active in O2 evolution compared with unwashed membranes. In the sensitivity of their O2-evolving apparatus to the composition of the medium in vitro, the red algae resembled blue-green algae and differed from other eukaryotic algae and higher plants. It is suggested that an environment of structured water may be essential for the functional integrity of Photosystem II in biliprotein-containing algae. PMID:6860312

Reconstitution of CF1-depleted thylakoid membranes with complete and fragmented chloroplast ATPase. The role of the delta subunit for proton conduction through CF0

NARCIS (Netherlands)

Engelbrecht, Siegfried; Lill, H; Junge, Wolfgang

1986-01-01

Chloroplast ATPase (CF1) was isolated from spinach, pea and maize thylakoids by EDTA extraction followed by anion-exchange chromatography. CF1 was purified and resolved by HPLC into integral CF1, and CF1 lacking the delta & epsilon subunits: CF1(-delta) and CF1(-epsilon). Washing Mono-Q-bound CF1

Protection of thylakoids against combined light and drought by a lumenal substance in the resurrection plant Haberlea rhodopensis

Science.gov (United States)

Georgieva, Katya; Sárvári, Éva; Keresztes, Áron

2010-01-01

Background and Aims Haberlea rhodopensis is a perennial, herbaceous, saxicolous, poikilohydric flowering plant that is able to survive desiccation to air-dried state under irradiance below 30 µmol m−2 s−1. However, desiccation at irradiance of 350 µmol m−2 s−1 induced irreversible changes in the photosynthetic apparatus, and mature leaves did not recover after rehydration. The aim here was to establish the causes and mechanisms of irreversible damage of the photosynthetic apparatus due to dehydration at high irradiance, and to elucidate the mechanisms determining recovery. Methods Changes in chloroplast structure, CO2 assimilation, chlorophyll fluorescence parameters, fluorescence imaging and the polypeptide patterns during desiccation of Haberlea under medium (100 µmol m−2 s−1; ML) irradiance were compared with those under low (30 µmol m−2 s−1; LL) irradiance. Key Results Well-watered plants (control) at 100 µmol m−2 s−1 were not damaged. Plants desiccated at LL or ML had similar rates of water loss. Dehydration at ML decreased the quantum efficiency of photosystem II photochemistry, and particularly the CO2 assimilation rate, more rapidly than at LL. Dehydration induced accumulation of stress proteins in leaves under both LL and ML. Photosynthetic activity and polypeptide composition were completely restored in LL plants after 1 week of rehydration, but changes persisted under ML conditions. Electron microscopy of structural changes in the chloroplast showed that the thylakoid lumen is filled with an electron-dense substance (dense luminal substance, DLS), while the thylakoid membranes are lightly stained. Upon dehydration and rehydration the DLS thinned and disappeared, the time course largely depending on the illumination: whereas DLS persisted during desiccation and started to disappear during late recovery under LL, it disappeared from the onset of dehydration and later was completely lost under ML. Conclusions Accumulation of DLS

Damage to photosystem II due to heat stress without light-driven electron flow: involvement of enhanced introduction of reducing power into thylakoid membranes.

Science.gov (United States)

Marutani, Yoko; Yamauchi, Yasuo; Kimura, Yukihiro; Mizutani, Masaharu; Sugimoto, Yukihiro

2012-08-01

Under a moderately heat-stressed condition, the photosystems of higher plants are damaged in the dark more easily than they are in the presence of light. To obtain a better understanding of this heat-derived damage mechanism that occurs in the dark, we focused on the involvement of the light-independent electron flow that occurs at 40 °C during the damage. In various plant species, the maximal photochemical quantum yield of photosystem (PS) II (Fv/Fm) decreased as a result of heat treatment in the dark. In the case of wheat, the most sensitive plant species tested, both Fv/Fm and oxygen evolution rapidly decreased by heat treatment at 40 °C for 30 min in the dark. In the damage, specific degradation of D1 protein was involved, as shown by immunochemical analysis of major proteins in the photosystem. Because light canceled the damage to PSII, the light-driven electron flow may play a protective role against PSII damage without light. Light-independent incorporation of reducing power from stroma was enhanced at 40 °C but not below 35 °C. Arabidopsis mutants that have a deficit of enzymes which mediate the incorporation of stromal reducing power into thylakoid membranes were tolerant against heat treatment at 40 °C in the dark, suggesting that the reduction of the plastoquinone pool may be involved in the damage. In conclusion, the enhanced introduction of reducing power from stroma into thylakoid membranes that occurs around 40 °C causes over-reduction of plastoquinone, resulting in the damage to D1 protein under heat stress without linear electron flow.

Consumption of thylakoid-rich spinach extract reduces hunger, increases satiety and reduces cravings for palatable food in overweight women.

Science.gov (United States)

Stenblom, Eva-Lena; Egecioglu, Emil; Landin-Olsson, Mona; Erlanson-Albertsson, Charlotte

2015-08-01

Green-plant membranes, thylakoids, have previously been found to increase postprandial release of the satiety hormone GLP-1, implicated in reward signaling. The purpose of this study was to investigate how treatment with a single dose of thylakoids before breakfast affects homeostatic as well as hedonic hunger, measured as wanting and liking for palatable food (VAS). We also examined whether treatment effects were correlated to scores for eating behavior. Compared to placebo, intake of thylakoids significantly reduced hunger (21% reduction, p snacks and sweets during the day (36% reduction, p snacks, respectively, and decreased subjective liking for sweet (28% reduction, p snacks, sweet-and-fat snacks in particular, were positively correlated to higher emotional eating scores (p obesity. Individuals scoring higher for emotional eating behavior may have enhanced treatment effect on cravings for palatable food. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

From isolated light-harvesting complexes to the thylakoid membrane: a single-molecule perspective

Science.gov (United States)

Gruber, J. Michael; Malý, Pavel; Krüger, Tjaart P. J.; Grondelle, Rienk van

2018-01-01

The conversion of solar radiation to chemical energy in plants and green algae takes place in the thylakoid membrane. This amphiphilic environment hosts a complex arrangement of light-harvesting pigment-protein complexes that absorb light and transfer the excitation energy to photochemically active reaction centers. This efficient light-harvesting capacity is moreover tightly regulated by a photoprotective mechanism called non-photochemical quenching to avoid the stress-induced destruction of the catalytic reaction center. In this review we provide an overview of single-molecule fluorescence measurements on plant light-harvesting complexes (LHCs) of varying sizes with the aim of bridging the gap between the smallest isolated complexes, which have been well-characterized, and the native photosystem. The smallest complexes contain only a small number (10-20) of interacting chlorophylls, while the native photosystem contains dozens of protein subunits and many hundreds of connected pigments. We discuss the functional significance of conformational dynamics, the lipid environment, and the structural arrangement of this fascinating nano-machinery. The described experimental results can be utilized to build mathematical-physical models in a bottom-up approach, which can then be tested on larger in vivo systems. The results also clearly showcase the general property of biological systems to utilize the same system properties for different purposes. In this case it is the regulated conformational flexibility that allows LHCs to switch between efficient light-harvesting and a photoprotective function.

Comparative study of thylakoid membranes in terminal heterocysts and vegetative cells from two cyanobacteria, Rivularia M-261 and Anabaena variabilis, by fluorescence and absorption spectral microscopy.

Science.gov (United States)

Nozue, Shuho; Katayama, Mitsunori; Terazima, Masahide; Kumazaki, Shigeichi

2017-09-01

Heterocyst is a nitrogen-fixing cell differentiated from a cell for oxygen-evolving photosynthesis (vegetative cell) in some filamentous cyanobacteria when fixed nitrogen (e.g., ammonia and nitrate) is limited. Heterocysts appear at multiple separated positions in a single filament with an interval of 10-20 cells in some genera (including Anabaena variabilis). In other genera, a single heterocyst appears only at the basal terminal in a filament (including Rivularia M-261). Such morphological diversity may necessitate different properties of heterocysts. However, possible differences in heterocysts have largely remained unexplored due to the minority of heterocysts among major vegetative cells. Here, we have applied spectroscopic microscopy to Rivularia and A. variabilis to analyze their thylakoid membranes in individual cells. Absorption and fluorescence spectral imaging enabled us to estimate concentrations and interconnections of key photosynthetic components like photosystem I (PSI), photosystem II (PSII) and subunits of light-harvesting phycobilisome including phycocyanin (PC). The concentration of PC in heterocysts of Rivularia is far higher than that of A. variabilis. Fluorescence quantum yield of PC in Rivularia heterocysts was found to be virtually the same as those in its vegetative cells, while fluorescence quantum yield of PC in A. variabilis heterocysts was enhanced in comparison with its vegetative cells. PSI concentration in the thylakoid membranes of heterocysts seems to remain nearly the same as those of the vegetative cells in both the species. The average stoichiometric ratio between PSI monomer and PC hexamer in Rivularia heterocysts is estimated to be about 1:1. Copyright © 2017 Elsevier B.V. All rights reserved.

Surface density and volume density measurements of chloroplast thylakoids in maize ( Zea mays L.) under chilling conditions

Czech Academy of Sciences Publication Activity Database

Kubínová, Lucie; Kutík, J.

2007-01-01

Roč. 45, č. 4 (2007), s. 481-488 ISSN 0300-3604 R&D Projects: GA AV ČR(CZ) IAA100110502; GA MŠk(CZ) LC06063 Grant - others:GA ČR(CZ) GA522/01/0846 Institutional research plan: CEZ:AV0Z50110509 Keywords : stereology * surface area * thylakoid membranes Subject RIV: EA - Cell Biology Impact factor: 0.976, year: 2007

JUMPING MODE ATOMIC FORCE MICROSCOPY ON GRANA MEMBRANES FROM SPINACH

NARCIS (Netherlands)

Sznee, K.; Dekker, J.P.; Dame, R.T.; Van Roon, H.; Wuite, G.J.L.; Frese, R.N.

2011-01-01

The thylakoid membrane system is a complex membrane system that organizes and reorganizes itself to provide plants optimal chemical energy from sunlight under different and varying environmental conditions. Grana membranes are part of this system and contain the light-driven water-splitting enzyme

A fluorescence detected magnetic resonance investigation of the carotenoid triplet states associated with Photosystem II of isolated spinach thylakoid membranes

CERN Document Server

Santabarbara, S; Carbonera, D; Heathcote, P

2005-01-01

The carotenoid triplet populations associated with the fluorescence emission chlorophyll forms of Photosystem II have been investigated in isolated spinach thylakoid membranes by means of fluorescence detected magnetic resonance in zero field (FDMR). The spectra collected in the 680-690 nm emission range, have been fitted by a global analysis procedure. At least five different carotenoid triplet states coupled to the terminal emitting chlorophyll forms of PS II, peaking at 682 nm, 687 nm and 692 nm, have been characterised. The triplets associated with the outer antenna emission forms, at 682 nm, have zero field splitting parameters D = 0.0385 cm/sup -1/, E = 0.00367 cm/sup -1/; D = 0.0404 cm/sup -1/, E = 0.00379 cm/sup -1/ and D = 0.0386 cm/sup -1/, E = 0.00406 cm/sup -1/ which are very similar to those previously reported for the xanthophylls of the isolated LHC II complex. Therefore the FDMR spectra recorded in this work provide insights into the organisation of the LHC II complex in the unperturbed enviro...

Progress in herbicide determination with the thylakoid bioassay.

Science.gov (United States)

Trapmann, S; Etxebarria, N; Schnabl, H; Grobecker, K H

1998-01-01

Chloroplast thylakoids are used as biological units to determine herbicides in different kinds of water samples as well as in aqueous extracts of compost, soil or food samples. The thylakoid bioassay shows clearly inhibition of fluorescence yield in the presence of photosystem II specific herbicides. Due to this method the ecotoxicological effect of samples with unknown pollutants can be tested fast and cost effective. It has been proven that all photosynthetic active compounds are recorded at the same time because only additive interactions occur. Therefore, the contamination level can be expressed as cumulative parameter for photosystem II active substances. Application was improved clearly by the addition of the radical scavenger sodium ascorbate to the isolation media and by a higher concentration of the measuring medium. A new data evaluation method is described yielding in a lower detection limit of 0.4 microg diuron/1. The guidelines for the quality of water for human consumption with an allowable concentration of pesticides in groups is 0,5 microg/1 and can be controlled with the thylakoid bioassay without performing any preconcentration steps.

Multilayer gyroid cubic membrane organization in green alga Zygnema.

Science.gov (United States)

Zhan, Ting; Lv, Wenhua; Deng, Yuru

2017-09-01

Biological cubic membranes (CM), which are fluid membranes draped onto the 3D periodic parallel surface geometries with cubic symmetry, have been observed within subcellular organelles, including mitochondria, endoplasmic reticulum, and thylakoids. CM transition tends to occur under various stress conditions; however, multilayer CM organizations often appear associated with light stress conditions. This report is about the characterization of a projected gyroid CM in a transmission electron microscopy study of the chloroplast membranes within green alga Zygnema (LB923) whose lamellar form of thylakoid membrane started to fold into multilayer gyroid CM in the culture at the end of log phase of cell growth. Using the techniques of computer simulation of transmission electron microscopy (TEM) and a direct template matching method, we show that these CM are based on the gyroid parallel surfaces. The single, double, and multilayer gyroid CM morphologies are observed in which space is continuously divided into two, three, and more subvolumes by either one, two, or several parallel membranes. The gyroid CM are continuous with varying amount of pseudo-grana with lamellar-like morphology. The relative amount and order of these two membrane morphologies seem to vary with the age of cell culture and are insensitive to ambient light condition. In addition, thylakoid gyroid CM continuously interpenetrates the pyrenoid body through stalk, bundle-like, morphologies. Inside the pyrenoid body, the membranes re-folded into gyroid CM. The appearance of these CM rearrangements due to the consequence of Zygnema cell response to various types of environmental stresses will be discussed. These stresses include nutrient limitation, temperature fluctuation, and ultraviolet (UV) exposure.

Research on the ultrafast fluorescence property of thylakoid membranes of the wild-type and mutant rice

Science.gov (United States)

Ren, Zhao-Yu; Xu, Xiao-Ming; Wang, Shui-Cai; Xin, Yue-Yong; He, Jun-Fang; Hou, Xun

2003-10-01

A high yielding rice variety mutant (Oryza sativa L., Zhenhui 249) with low chlorophyll b (Chl b) has been discovered in natural fields. It has a quality character controlled by a pair of recessive genes (nuclear gene). The partial loss of Chl b in content affects the efficiency of light harvest in a light harvest complex (LHC), thus producing the difference of the exciting energy transfer and the efficiency of photochemistry conversion between the mutant and wild-type rice in photosynthetic unit. The efficiency of utilizing light energy is higher in the mutant than that in the wild-type rice relatively. For further discussion of the above-mentioned difference and learning about the mechanism of the increase in the photochemical efficiency of the mutant, the pico-second resolution fluorescence spectrum measurement with delay-frame-scanning single photon counting technique is adopted. Thylakoid membranes of the mutant and the wild-type rice are excited by an Ar+ laser with a pulse width of 120 ps, repetition rate of 4 MHz and wavelength of 514 nm. Compared with the time and spectrum property of exciting fluorescence, conclusions of those ultrafast dynamic experiments are: 1) The speeds of the exciting energy transferred in photo-system I are faster than that in photo-system II in both samples. 2) The speeds of the exciting energy transfer of mutant sample are faster than those of the wild-type. This might be one of the major reasons why the efficiency of photosynthesis is higher in mutant than that in the wild-type rice.

Membrane Stored Curvature Elastic Stress Modulates Recruitment of Maintenance Proteins PspA and Vipp1.

Science.gov (United States)

McDonald, Christopher; Jovanovic, Goran; Ces, Oscar; Buck, Martin

2015-09-01

Phage shock protein A (PspA), which is responsible for maintaining inner membrane integrity under stress in enterobacteria, and vesicle-inducting protein in plastids 1 (Vipp1), which functions for membrane maintenance and thylakoid biogenesis in cyanobacteria and plants, are similar peripheral membrane-binding proteins. Their homologous N-terminal amphipathic helices are required for membrane binding; however, the membrane features recognized and required for expressing their functionalities have remained largely uncharacterized. Rigorously controlled, in vitro methodologies with lipid vesicles and purified proteins were used in this study and provided the first biochemical and biophysical characterizations of membrane binding by PspA and Vipp1. Both proteins are found to sense stored curvature elastic (SCE) stress and anionic lipids within the membrane. PspA has an enhanced sensitivity for SCE stress and a higher affinity for the membrane than Vipp1. These variations in binding may be crucial for some of the proteins' differing roles in vivo. Assays probing the transcriptional regulatory function of PspA in the presence of vesicles showed that a relief of transcription inhibition occurs in an SCE stress-specific manner. This in vitro recapitulation of membrane stress-dependent transcription control suggests that the Psp response may be mounted in vivo when a cell's inner membrane experiences increased SCE stress. All cell types maintain the integrity of their membrane systems. One widely distributed membrane stress response system in bacteria is the phage shock protein (Psp) system. The central component, peripheral membrane protein PspA, which mitigates inner membrane stress in bacteria, has a counterpart, Vipp1, which functions for membrane maintenance and thylakoid biogenesis in plants and photosynthetic bacteria. Membrane association of both these proteins is accepted as playing a pivotal role in their functions. Here we show that direct membrane binding by

Reversible membrane reorganizations during photosynthesis in vivo: revealed by small-angle neutron scattering

DEFF Research Database (Denmark)

Nagy, Gergely; Posselt, Dorthe; Kovacs, Laszlo

2011-01-01

In the present study, we determined characteristic repeat distances of the photosynthetic membranes in living cyanobacterial and eukaryotic algal cells, and in intact thylakoid membranes isolated from higher plants with time-resolved small-angle neutron scattering. This non-invasive technique...

Hacking the thylakoid proton motive force for improved photosynthesis: modulating ion flux rates that control proton motive force partitioning into Δψ and ΔpH.

Science.gov (United States)

Davis, Geoffry A; Rutherford, A William; Kramer, David M

2017-09-26

There is considerable interest in improving plant productivity by altering the dynamic responses of photosynthesis in tune with natural conditions. This is exemplified by the 'energy-dependent' form of non-photochemical quenching ( q E ), the formation and decay of which can be considerably slower than natural light fluctuations, limiting photochemical yield. In addition, we recently reported that rapidly fluctuating light can produce field recombination-induced photodamage (FRIP), where large spikes in electric field across the thylakoid membrane (Δ ψ ) induce photosystem II recombination reactions that produce damaging singlet oxygen ( 1 O 2 ). Both q E and FRIP are directly linked to the thylakoid proton motive force ( pmf ), and in particular, the slow kinetics of partitioning pmf into its ΔpH and Δ ψ components. Using a series of computational simulations, we explored the possibility of 'hacking' pmf partitioning as a target for improving photosynthesis. Under a range of illumination conditions, increasing the rate of counter-ion fluxes across the thylakoid membrane should lead to more rapid dissipation of Δ ψ and formation of ΔpH. This would result in increased rates for the formation and decay of q E while resulting in a more rapid decline in the amplitudes of Δ ψ -spikes and decreasing 1 O 2 production. These results suggest that ion fluxes may be a viable target for plant breeding or engineering. However, these changes also induce transient, but substantial mismatches in the ATP : NADPH output ratio as well as in the osmotic balance between the lumen and stroma, either of which may explain why evolution has not already accelerated thylakoid ion fluxes. Overall, though the model is simplified, it recapitulates many of the responses seen in vivo , while spotlighting critical aspects of the complex interactions between pmf components and photosynthetic processes. By making the programme available, we hope to enable the community of photosynthesis

Modeling the light-induced electric potential difference (ΔΨ), the pH difference (ΔpH) and the proton motive force across the thylakoid membrane in C3 leaves.

Science.gov (United States)

Lyu, Hui; Lazár, Dušan

2017-01-21

A model was constructed which includes electron transport (linear and cyclic and Mehler type reaction) coupled to proton translocation, counter ion movement, ATP synthesis, and Calvin-Benson cycle. The focus is on modeling of the light-induced total electric potential difference (ΔΨ) which in this model originates from the bulk phase electric potential difference (ΔΨ b ), the localized electric potential difference (ΔΨ c ), as well as the surface electric potential difference (ΔΨ s ). The measured dual wavelength transmittance signal (ΔA515-560nm, electrochromic shift) was used as a proxy for experimental ΔΨ. The predictions for theoretical ΔΨ vary with assumed contribution of ΔΨ s , which might imply that the measured ΔA515-560nm trace on a long time scale reflects the interplay of the ΔΨ components. Simulations also show that partitioning of proton motive force (pmf) to ΔΨ b and ΔpH components is sensitive to the stoichiometric ratio of H + /ATP, energy barrier for ATP synthesis, ionic strength, buffer capacity and light intensity. Our model shows that high buffer capacity promotes the establishment of ΔΨ b , while the formation of pH i minimum is not 'dissipated' but 'postponed' until it reaches the same level as that for low buffer capacity. Under physiologically optimal conditions, the output of the model shows that at steady state in light, the ΔpH component is the main contributor to pmf to drive ATP synthesis while a low ΔΨ b persists energizing the membrane. Our model predicts 11mV as the resting electric potential difference across the thylakoid membrane in dark. We suggest that the model presented in this work can be integrated as a module into a more comprehensive model of oxygenic photosynthesis. Copyright © 2016 Elsevier Ltd. All rights reserved.

Dietary green-plant thylakoids decrease gastric emptying and gut transit, promote changes in the gut microbial flora, but does not cause steatorrhea

DEFF Research Database (Denmark)

Stenblom, Eva-Lena; Weström, Björn R.; Linninge, Caroline

2016-01-01

Green-plant thylakoids increase satiety by affecting appetite hormones such as ghrelin, cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1). The objective of this study was to investigate if thylakoids also affect gastrointestinal (GI) passage and microbial composition. To analyse the effects......, and specifically the Bacteriodes fragilis group, were increased by thylakoid treatment versus placebo, while thylakoids did not cause steatorrhea. Dietary supplementation with thylakoids thus affects satiety both via appetite hormones and GI fullness, and affects the microbial composition without causing GI...... adverse effects such as steatorrhea. This suggests thylakoids as a novel agent in prevention and treatment of obesity....

Ultrastructural changes in the membrane system of isolated chloroplasts of spinach under the influence of carbonic anhydrase inhibitors AA and EA

Directory of Open Access Journals (Sweden)

Marina V. Vodka

2013-04-01

Full Text Available The effects of carbonic anhydrase inhibitors (АА and EA on the membrane system of isolated chloroplasts of spinach were investigated. Under the influence of AA the considerable alterations in granal structure occurred, the thickness of the granal thylakoids increased by 36% and the interspace between thylakoids by 10% comparable with the control. As a result of EA treatment, the thickness of granal thylakoids enhanced by 31% and the interspace between thylakoids increased by 8% in comparison to the control. It was shown that structure of the granal system of the chloroplast was more sensitive to AA than EA. The data obtained can indicate a decrease in the activity of the thylakoid carbonic anhydrase, inhibition of electron transport and photosynthetic process as a whole in the presence of carbonic anhydrase inhibitors (AA and EA.

Structure and membrane organization of photosystem II in green plants

NARCIS (Netherlands)

Hankamer, B; Barber, J; Boekema, EJ

1997-01-01

Photosystem II (PSII) is the pigment protein complex embedded in the thylakoid membrane of higher plants, algae, and cyanobacteria that uses solar energy to drive the photosynthetic water-splitting reaction. This chapter reviews the primary, secondary, tertiary, and quaternary structures of PSII as

In vitro degradation of the 32kDa PS II reaction centre protein

International Nuclear Information System (INIS)

Eckenswiller, L.C.; Greenberg, B.M.

1989-01-01

The 32kDa thylakoid membrane protein is an integral component of the PS II reaction centre. The protein, although stable in the dark, undergoes light dependent turnover. Light from the UV, visible and far-red spectral regions induce 32kDa protein degradation. To better understand 32kDa protein metabolism, an in vitro degradation system is being developed. It consists of isolated thylakoid membranes than contain radiolabelled protein. The 32kDa protein is actively and specifically degraded when the thylakoid preparation is exposed to UV or visible radiation. The protein is stable in the dark. The herbicides (atrazine and DCMU) inhibit degradation in the in vitro system as they do in vivo. Additionally, several methods of isolating thylakoids are being compared to optimize the 32kDa protein degradation reaction. The preparations will be evaluated based on their ability to permit light dependent degradation of the 32kDa protein without affecting the other membrane components

Consequences of C4 differentiation for chloroplast membrane proteomes in maize mesophyll and bundle sheath cells.

Science.gov (United States)

Majeran, Wojciech; Zybailov, Boris; Ytterberg, A Jimmy; Dunsmore, Jason; Sun, Qi; van Wijk, Klaas J

2008-09-01

Chloroplasts of maize leaves differentiate into specific bundle sheath (BS) and mesophyll (M) types to accommodate C(4) photosynthesis. Chloroplasts contain thylakoid and envelope membranes that contain the photosynthetic machineries and transporters but also proteins involved in e.g. protein homeostasis. These chloroplast membranes must be specialized within each cell type to accommodate C(4) photosynthesis and regulate metabolic fluxes and activities. This quantitative study determined the differentiated state of BS and M chloroplast thylakoid and envelope membrane proteomes and their oligomeric states using innovative gel-based and mass spectrometry-based protein quantifications. This included native gels, iTRAQ, and label-free quantification using an LTQ-Orbitrap. Subunits of Photosystems I and II, the cytochrome b(6)f, and ATP synthase complexes showed average BS/M accumulation ratios of 1.6, 0.45, 1.0, and 1.33, respectively, whereas ratios for the light-harvesting complex I and II families were 1.72 and 0.68, respectively. A 1000-kDa BS-specific NAD(P)H dehydrogenase complex with associated proteins of unknown function containing more than 15 proteins was observed; we speculate that this novel complex possibly functions in inorganic carbon concentration when carboxylation rates by ribulose-bisphosphate carboxylase/oxygenase are lower than decarboxylation rates by malic enzyme. Differential accumulation of thylakoid proteases (Egy and DegP), state transition kinases (STN7,8), and Photosystem I and II assembly factors was observed, suggesting that cell-specific photosynthetic electron transport depends on post-translational regulatory mechanisms. BS/M ratios for inner envelope transporters phosphoenolpyruvate/P(i) translocator, Dit1, Dit2, and Mex1 were determined and reflect metabolic fluxes in carbon metabolism. A wide variety of hundreds of other proteins showed differential BS/M accumulation. Mass spectral information and functional annotations are

Impact of in vitro cultivation conditions on stress responses and on changes in thylakoid membrane proteins and pigments of tobacco during ex vitro acclimation

Czech Academy of Sciences Publication Activity Database

Hofman, Petr; Haisel, Daniel; Komenda, J.; Vágner, Martin; Tichá, I.; Schäfer, C.; Čapková, Věra

2002-01-01

Roč. 45, č. 2 (2002), s. 189-195 ISSN 0006-3134 R&D Projects: GA MŠk OC 843.50; GA AV ČR IAA5038207 Institutional research plan: CEZ:AV0Z5038910 Keywords : thylakoid * proteins * tobacco Subject RIV: EF - Botanics Impact factor: 0.583, year: 2002

Composite membrane with integral rim

Science.gov (United States)

Routkevitch, Dmitri; Polyakov, Oleg G

2015-01-27

Composite membranes that are adapted for separation, purification, filtration, analysis, reaction and sensing. The composite membranes can include a porous support structure having elongate pore channels extending through the support structure. The composite membrane also includes an active layer comprising an active layer material, where the active layer material is completely disposed within the pore channels between the surfaces of the support structure. The active layer is intimately integrated within the support structure, thus enabling great robustness, reliability, resistance to mechanical stress and thermal cycling, and high selectivity. Methods for the fabrication of composite membranes are also provided.

Integrable structure in discrete shell membrane theory.

Science.gov (United States)

Schief, W K

2014-05-08

We present natural discrete analogues of two integrable classes of shell membranes. By construction, these discrete shell membranes are in equilibrium with respect to suitably chosen internal stresses and external forces. The integrability of the underlying equilibrium equations is proved by relating the geometry of the discrete shell membranes to discrete O surface theory. We establish connections with generalized barycentric coordinates and nine-point centres and identify a discrete version of the classical Gauss equation of surface theory.

Kinetics of structural reorganizations in multilamellarphotosynthetic membranes monitored by small-angle neutronscattering

DEFF Research Database (Denmark)

Nagy, Gergely; Kovacs, Laszlo; Unnep, Renata

2013-01-01

and in unicellular organisms, we discuss the advantages and technical and methodological limitations of timeresolved SANS. We present a detailed and more systematical investigation of the kinetics of light-induced structural reorganizations in isolated spinach thylakoid membranes, which show how changes...

Fine structure of granal thylakoid membrane organization using cryo electron tomography

NARCIS (Netherlands)

Kouril, Roman; Oostergetel, Gert T.; Boekema, Egbert J.

The architecture of grana membranes from spinach chloroplasts was studied by cryo electron tomography. Tomographic reconstructions of ice-embedded isolated grana stacks enabled to resolve features of photosystem II (PSII) in the native membrane and to assign the absolute orientation of individual

Dietary thylakoids suppress blood glucose and modulate appetite-regulating hormones in pigs exposed to oral glucose tolerance test

DEFF Research Database (Denmark)

Montelius, Caroline; Szwiec, Katarzyna; Kardas, Marek

2014-01-01

BACKGROUND & AIMS: Dietary chloroplast thylakoids have previously been found to reduce food intake and body weight in animal models, and to change metabolic profiles in humans in mixed-food meal studies. The aim of this study was to investigate the modulatory effects of thylakoids on glucose...... metabolism and appetite-regulating hormones during an oral glucose tolerance test in pigs fed a high fat diet. METHODS: Six pigs were fed a high fat diet (36 energy% fat) for one month before oral glucose tolerance test (1 g/kg d-glucose) was performed. The experiment was designed as a cross-over study......, either with or without addition of 0.5 g/kg body weight of thylakoid powder. RESULTS: The supplementation of thylakoids to the oral glucose tolerance test resulted in decreased blood glucose concentrations during the first hour, increased plasma cholecystokinin concentrations during the first two hours...

Characterization of the cytochrome c oxidase in isolated and purified plasma membranes from the cyanobacterium Anacystis nidulans

International Nuclear Information System (INIS)

Peschek, G.A.; Wastyn, M.; Trnka, M.; Molitor, V.; Fry, I.V.; Packer, L.

1989-01-01

Functionally intact plasma membranes were isolated from the cyanobacterium (blue-green alga) Anacystis nidulans through French pressure cell extrusion of lysozyme/EDTA-treated cells, separated from thylakoid membranes by discontinuous sucrose density gradient centrifugation, and purified by repeated recentrifugation. Origin and identity of the chlorophyll-free plasma membrane fraction were confirmed by labeling of intact cells with impermeant protein markers, [ 35 S]diazobenzenesulfonate and fluorescamine, prior to membrane isolation. Rates of oxidation of reduced horse heart cytochrome c by purified plasma and thylakoid membranes were 90 and 2 nmol min -1 (mg of protein) -1 , respectively. The cytochrome oxidase in isolated plasma membranes was identified as a copper-containing aa 3 -type enzyme from the properties of its redox-active and EDTA-resistant Cu 2+ ESR signal, the characteristic inhibition profile, reduced minus oxidized difference spectra, carbon monoxide difference spectra, photoaction and photodissociation spectra of the CO-inhibited enzyme, and immunological cross-reaction of two subunits of the enzyme with antibodies against subunits I and II, and the holoenzyme, of Paracoccus denitrificans aa 3 -type cytochrome oxidase. The data presented are the first comprehensive evidence for the occurrence of aa 3 -type cytochrome oxidase in the plasma membrane of a cyanobacterium similar to the corresponding mitochondrial enzyme

Treating domestic sewage by Integrated Inclined-Plate-Membrane bio-reactor

Science.gov (United States)

Song, Li Ming; Wang, Zi; Chen, Lei; Zhong, Min; Dong, Zhan Feng

2017-12-01

Membrane fouling shorten the service life of the membrane and increases aeration rate for membrane surface cleaning. Two membrane bio-reactors, one for working and another for comparing, were set up to evaluate the feasibility of alleviating membrane fouling and improving wastewater treatment efficiency by integrating inclined-plate precipitation and membrane separation. The result show that: (1) Inclined-plate in reactor had a good effect on pollutant removal of membrane bioreactor. The main role of inclined-plate is dividing reactor space and accelerating precipitation. (2) Working reactor have better performance in COD, TN and TP removal, which can attribute to that working reactor (integrated inclined-plate-Membrane bioreactor) takes both advantages of membrane separation and biological treatment. When influent COD, TP and TN concentration is 163-248 mg/L, 2.08-2.81 mg/L and 24.38-30.49 mg/L in working reactor, effluent concentration is 27-35 mg/L, 0.53-0.59 mg/L and 11.28-11.56 mg/L, respectively. (3) Membrane fouling was well alleviated in integrated inclined-plate-Membrane bioreactor, and membrane normal service time is significantly longer than that in comparing reactor, which can attribute to accelerating precipitation of inclined-plate. In summary, integrated inclined-plate-Membrane bioreactor is a promising technology to alleviating membrane fouling and improving wastewater treatment efficiency, having good performance and bright future in application.

Neutrophil glycoprotein Mo1 is an integral membrane protein of plasma membranes and specific granules

International Nuclear Information System (INIS)

Stevenson, K.B.; Nauseef, W.M.; Clark, R.A.

1987-01-01

The glucoprotein Mo1 has previously been demonstrated to be on the cell surface and in the specific granule fraction of neutrophils and to be translocated to the cell surface during degranulation. It is not known, however, whether Mo1 is an integral membrane protein or a soluble, intragranular constituent loosely associated with the specific granule membrane. Purified neutrophils were disrupted by nitrogen cavitation and separated on Percoll density gradients into four fractions enriched for azurophilic granules, specific granules, plasma membrane, and cytosol, respectively. The glycoproteins in these fractions were labeled with 3 H-borohydride reduction, extracted with Triton X-114, and immunoprecipitated with 60.3, an anti-Mo1 monoclonal antibody. Mo1 was detected only in the specific granule and plasma membrane fractions and partitioned exclusively into the detergent-rich fraction consistent with Mo1 being an integral membrane protein. In addition, treatment of specific granule membranes with a high salt, high urea buffer to remove adsorbed or peripheral proteins failed to dissociate Mo1. These data support the hypothesis that Mo1 is an integral membrane protein of plasma and specific granule membranes in human neutrophils

Organic membrane photonic integrated circuits (OMPICs).

Science.gov (United States)

Amemiya, Tomohiro; Kanazawa, Toru; Hiratani, Takuo; Inoue, Daisuke; Gu, Zhichen; Yamasaki, Satoshi; Urakami, Tatsuhiro; Arai, Shigehisa

2017-08-07

We propose the concept of organic membrane photonic integrated circuits (OMPICs), which incorporate various functions needed for optical signal processing into a flexible organic membrane. We describe the structure of several devices used within the proposed OMPICs (e.g., transmission lines, I/O couplers, phase shifters, photodetectors, modulators), and theoretically investigate their characteristics. We then present a method of fabricating the photonic devices monolithically in an organic membrane and demonstrate the operation of transmission lines and I/O couplers, the most basic elements of OMPICs.

Structural adaptations of proteins to different biological membranes

Science.gov (United States)

Pogozheva, Irina D.; Tristram-Nagle, Stephanie; Mosberg, Henry I.; Lomize, Andrei L.

2013-01-01

To gain insight into adaptations of proteins to their membranes, intrinsic hydrophobic thicknesses, distributions of different chemical groups and profiles of hydrogen-bonding capacities (α and β) and the dipolarity/polarizability parameter (π*) were calculated for lipid-facing surfaces of 460 integral α-helical, β-barrel and peripheral proteins from eight types of biomembranes. For comparison, polarity profiles were also calculated for ten artificial lipid bilayers that have been previously studied by neutron and X-ray scattering. Estimated hydrophobic thicknesses are 30-31 Å for proteins from endoplasmic reticulum, thylakoid, and various bacterial plasma membranes, but differ for proteins from outer bacterial, inner mitochondrial and eukaryotic plasma membranes (23.9, 28.6 and 33.5 Å, respectively). Protein and lipid polarity parameters abruptly change in the lipid carbonyl zone that matches the calculated hydrophobic boundaries. Maxima of positively charged protein groups correspond to the location of lipid phosphates at 20-22 Å distances from the membrane center. Locations of Tyr atoms coincide with hydrophobic boundaries, while distributions maxima of Trp rings are shifted by 3-4 Å toward the membrane center. Distributions of Trp atoms indicate the presence of two 5-8 Å-wide midpolar regions with intermediate π* values within the hydrocarbon core, whose size and symmetry depend on the lipid composition of membrane leaflets. Midpolar regions are especially asymmetric in outer bacterial membranes and cell membranes of mesophilic but not hyperthermophilic archaebacteria, indicating the larger width of the central nonpolar region in the later case. In artificial lipid bilayers, midpolar regions are observed up to the level of acyl chain double bonds. PMID:23811361

Flip-flop of phospholipids in proteoliposomes reconstituted from detergent extract of chloroplast membranes: kinetics and phospholipid specificity.

Directory of Open Access Journals (Sweden)

Archita Rajasekharan

Full Text Available Eukaryotic cells are compartmentalized into distinct sub-cellular organelles by lipid bilayers, which are known to be involved in numerous cellular processes. The wide repertoire of lipids, synthesized in the biogenic membranes like the endoplasmic reticulum and bacterial cytoplasmic membranes are initially localized in the cytosolic leaflet and some of these lipids have to be translocated to the exoplasmic leaflet for membrane biogenesis and uniform growth. It is known that phospholipid (PL translocation in biogenic membranes is mediated by specific membrane proteins which occur in a rapid, bi-directional fashion without metabolic energy requirement and with no specificity to PL head group. A recent study reported the existence of biogenic membrane flippases in plants and that the mechanism of plant membrane biogenesis was similar to that found in animals. In this study, we demonstrate for the first time ATP independent and ATP dependent flippase activity in chloroplast membranes of plants. For this, we generated proteoliposomes from Triton X-100 extract of intact chloroplast, envelope membrane and thylakoid isolated from spinach leaves and assayed for flippase activity using fluorescent labeled phospholipids. Half-life time of flipping was found to be 6 ± 1 min. We also show that: (a intact chloroplast and envelope membrane reconstituted proteoliposomes can flip fluorescent labeled analogs of phosphatidylcholine in ATP independent manner, (b envelope membrane and thylakoid reconstituted proteoliposomes can flip phosphatidylglycerol in ATP dependent manner, (c Biogenic membrane ATP independent PC flipping activity is protein mediated and (d the kinetics of PC translocation gets affected differently upon treatment with protease and protein modifying reagents.

Fe deficiency induced changes in rice (Oryza sativa L.) thylakoids.

Science.gov (United States)

Wang, Yuwen; Xu, Chao; Li, Kang; Cai, Xiaojie; Wu, Min; Chen, Guoxiang

2017-01-01

Iron deficiency is an important abiotic stress that limits productivity of crops all over the world. We selected a hybrid rice (Oryza sativa L.), LYPJ, which is super high-yield and widely cultured in China, to investigate changes in the components and structure of thylakoid membranes and photosynthetic performance in response to iron deficiency. Our results demonstrated that photosystem I (PSI) is the primary target for iron deficiency, while the changes in photosystem II (PSII) are important for rebuilding a balance in disrupted energy utilization and dissipation caused by differential degradation of photosynthetic components. The result of immunoblot analysis suggested that the core subunit PsaA declined drastically, while PsbA remained relatively stable. Furthermore, several organizational changes of the photosynthetic apparatus were found by BN-PAGE, including a marked decrease in the PSI core complexes, the Cytb 6 /f complex, and the trimeric form of the LHCII antenna, consistent with the observed unstacking grana. The fluorescence induction analysis indicated a descending PSII activity with energy dissipation enhanced markedly. In addition, we proposed that the crippled CO 2 assimilation could be compensated by the enhanced of phosphoenolpyruvate carboxylase (PEPC), which is suggested by the decreased ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) and photosynthetic efficiency.

Novel protein phosphorylation site identification in spinach stroma membranes by titanium dioxide microcolumns and tandem mass spectrometry

DEFF Research Database (Denmark)

Rinalducci, Sara; Larsen, Martin Røssel; Mohammed, Shabaz

2006-01-01

In this work, spinach stroma membrane, instead of thylakoid, has been investigated for the presence of phosphorylated proteins. We identified seven previously unknown phosphorylation sites by taking advantage of TiO(2) phosphopeptides enrichment coupled to mass spectrometric analysis. Upon...

An Integrated Membrane Process for Butenes Production

Directory of Open Access Journals (Sweden)

Leonardo Melone

2016-11-01

Full Text Available Iso-butene is an important material for the production of chemicals and polymers. It can take part in various chemical reactions, such as hydrogenation, oxidation and other additions owing to the presence of a reactive double bond. It is usually obtained as a by-product of a petroleum refinery, by Fluidized Catalytic Cracking (FCC of naphtha or gas-oil. However, an interesting alternative to iso-butene production is n-butane dehydroisomerization, which allows the direct conversion of n-butane via dehydrogenation and successive isomerization. In this work, a simulation analysis of an integrated membrane system is proposed for the production and recovery of butenes. The dehydroisomerization of n-butane to iso-butene takes place in a membrane reactor where the hydrogen is removed from the reaction side with a Pd/Ag alloys membrane. Afterwards, the retentate and permeate post-processing is performed in membrane separation units for butenes concentration and recovery. Four different process schemes are developed. The performance of each membrane unit is analyzed by appropriately developed performance maps, to identify the operating conditions windows and the membrane permeation properties required to maximize the recovery of the iso-butene produced. An analysis of integrated systems showed a yield of butenes higher than the other reaction products with high butenes recovery in the gas separation section, with values of molar concentration between 75% and 80%.

PSB27: A thylakoid protein enabling Arabidopsis to adapt to changing light intensity

Energy Technology Data Exchange (ETDEWEB)

Hou, Xin [Univ. of California, Berkeley, CA (United States). Dept of Plant and Microbial Biology; Garcia, Veder J. [Univ. of California, Berkeley, CA (United States). Dept of Plant and Microbial Biology; Buchanan, Bob B. [Univ. of California, Berkeley, CA (United States). Dept of Plant and Microbial Biology; Luan, Sheng [Univ. of California, Berkeley, CA (United States). Dept of Plant and Microbial Biology

2016-08-22

dissect the mechanism of action for CYP38. Our studies suggest an “autoinhibitory” model for CYP38 action. We plan to test this model and determine the mechanistic details for CYP38 function by biochemical and genetic procedures. 2. To determine the mechanism of action of FKBP20-2/FKBP16-3. Studies on FBKP16-3 and FKBP20-2 suggest that they may work together as a redox-dependent duo in PSII assembly. We will test this hypothesis using biochemical and genetic tools. 3. To determine the function of lumenal FKBPs by multi-gene mutagenesis approach. Using new genetic knockout (KO) procedures especially CRISPR/CAS9 system, we plan to generate multi-gene KO models that will likely provide vital information on those FKBPs with functional redundancy. 4. Functional analysis of thylakoid lumen network. We will focus our effort on objectives 1-3. Objective 4 represents a long-term goal to establish a more comprehensive network of proteins in the thylakoid lumen and their function in the assembly and maintenance of photosynthetic complexes. I hypothesize that the 16 immunophilin-type chaperones and foldases in the thylakoid lumen constitute an “assembly line” for the various components in the photosynthetic electron transport chain. Understanding this assembly line will enable further engineering of more efficient photosynthetic machines to capture more light energy and enhance plant productivity. Furthermore, it is envisioned that such molecular “assembly line” may be rebuilt to produce artificial photosynthesis in vitro or in non-photosynthetic organisms. These are highly relevant to the mission of “Photosynthetic Systems” and “Physical Biosciences” programs to “enhance our understanding of energy capture, conversion, and/or storage.” During the funding period, we have performed a number of experiments under the proposed objectives and obtained several new results. We have found in Objective 1 that CYP38 is associated to the thylokoid membrane with a larger

InP membrane on silicon integration technology

NARCIS (Netherlands)

Smit, M.K.

2013-01-01

Integration of light sources in silicon photonics is usually done with an active InP-based layer stack on a silicon-based photonic circuit-layer. InP Membrane On Silicon (IMOS) technology integrates all functionality in a single InP-based layer.

Excitation energy transfer between Light-harvesting complex II and Photosystem I in reconstituted membranes.

Science.gov (United States)

Akhtar, Parveen; Lingvay, Mónika; Kiss, Teréz; Deák, Róbert; Bóta, Attila; Ughy, Bettina; Garab, Győző; Lambrev, Petar H

2016-04-01

Light-harvesting complex II (LHCII), the major peripheral antenna of Photosystem II in plants, participates in several concerted mechanisms for regulation of the excitation energy and electron fluxes in thylakoid membranes. In part, these include interaction of LHCII with Photosystem I (PSI) enhancing the latter's absorption cross-section - for example in the well-known state 1 - state 2 transitions or as a long-term acclimation to high light. In this work we examined the capability of LHCII to deliver excitations to PSI in reconstituted membranes in vitro. Proteoliposomes with native plant thylakoid membrane lipids and different stoichiometric ratios of LHCII:PSI were reconstituted and studied by steady-state and time-resolved fluorescence spectroscopy. Fluorescence emission from LHCII was strongly decreased in PSI-LHCII membranes due to trapping of excitations by PSI. Kinetic modelling of the time-resolved fluorescence data revealed the existence of separate pools of LHCII distinguished by the time scale of energy transfer. A strongly coupled pool, equivalent to one LHCII trimer per PSI, transferred excitations to PSI with near-unity efficiency on a time scale of less than 10ps but extra LHCIIs also contributed significantly to the effective antenna size of PSI, which could be increased by up to 47% in membranes containing 3 LHCII trimers per PSI. The results demonstrate a remarkable competence of LHCII to increase the absorption cross-section of PSI, given the opportunity that the two types of complexes interact in the membrane. Copyright © 2016 Elsevier B.V. All rights reserved.

Integration of lateral porous silicon membranes into planar microfluidics.

Science.gov (United States)

Leïchlé, Thierry; Bourrier, David

2015-02-07

In this work, we present a novel fabrication process that enables the monolithic integration of lateral porous silicon membranes into single-layer planar microchannels. This fabrication technique relies on the patterning of local electrodes to guide pore formation horizontally within the membrane and on the use of silicon-on-insulator substrates to spatially localize porous silicon within the channel depth. The feasibility of our approach is studied by current flow analysis using the finite element method and supported by creating 10 μm long mesoporous membranes within 20 μm deep microchannels. The fabricated membranes are demonstrated to be potentially useful for dead-end microfiltration by adequately retaining 300 nm diameter beads while macromolecules such as single-stranded DNA and immunoglobulin G permeate the membrane. The experimentally determined fluidic resistance is in accordance with the theoretical value expected from the estimated pore size and porosity. The work presented here is expected to greatly simplify the integration of membranes capable of size exclusion based separation into fluidic devices and opens doors to the use of porous silicon in planar lab on a chip devices.

Impact of the antimicrobial peptide Novicidin on membrane structure and integrity

DEFF Research Database (Denmark)

Nielsen, Søren B; Otzen, Daniel Erik

2010-01-01

We have studied the impact of an 18-residue cationic antimicrobial peptide Novicidin (Nc) on the structure and integrity of partially anionic lipid membranes using oriented circular dichroism (OCD), quartz crystal microbalance with dissipation (QCM-D), dual polarization interferometry (DPI......), calcein dye leakage and fluorescence spectroscopy. OCD consistently showed that Nc is bound in an alpha-helical, surface bound state over a range of peptide to lipid (P/L) ratios up to approximately 1:15. Realignment of Nc at higher P/L ratios correlates to loss of membrane integrity as shown by Laurdan...... concentration, probably through formation of transient pores or transient disruption of the membrane integrity, followed by more extensive membrane disintegration at higher P/L ratios....

Free Flow Zonal Electrophoresis for Fractionation of Plant Membrane Compartments Prior to Proteomic Analysis.

Science.gov (United States)

Barkla, Bronwyn J

2018-01-01

Free flow zonal electrophoresis (FFZE) is a versatile, reproducible, and potentially high-throughput technique for the separation of plant organelles and membranes by differences in membrane surface charge. It offers considerable benefits over traditional fractionation techniques, such as density gradient centrifugation and two-phase partitioning, as it is relatively fast, sample recovery is high, and the method provides unparalleled sample purity. It has been used to successfully purify chloroplasts and mitochondria from plants but also, to obtain highly pure fractions of plasma membrane, tonoplast, ER, Golgi, and thylakoid membranes. Application of the technique can significantly improve protein coverage in large-scale proteomics studies by decreasing sample complexity. Here, we describe the method for the fractionation of plant cellular membranes from leaves by FFZE.

Anionic lipids and the cytoskeletal proteins MreB and RodZ define the spatio-temporal distribution and function of membrane stress controller PspA in Escherichia coli.

Science.gov (United States)

Jovanovic, Goran; Mehta, Parul; Ying, Liming; Buck, Martin

2014-11-01

All cell types must maintain the integrity of their membranes. The conserved bacterial membrane-associated protein PspA is a major effector acting upon extracytoplasmic stress and is implicated in protection of the inner membrane of pathogens, formation of biofilms and multi-drug-resistant persister cells. PspA and its homologues in Gram-positive bacteria and archaea protect the cell envelope whilst also supporting thylakoid biogenesis in cyanobacteria and higher plants. In enterobacteria, PspA is a dual function protein negatively regulating the Psp system in the absence of stress and acting as an effector of membrane integrity upon stress. We show that in Escherichia coli the low-order oligomeric PspA regulatory complex associates with cardiolipin-rich, curved polar inner membrane regions. There, cardiolipin and the flotillin 1 homologue YqiK support the PspBC sensors in transducing a membrane stress signal to the PspA-PspF inhibitory complex. After stress perception, PspA high-order oligomeric effector complexes initially assemble in polar membrane regions. Subsequently, the discrete spatial distribution and dynamics of PspA effector(s) in lateral membrane regions depend on the actin homologue MreB and the peptidoglycan machinery protein RodZ. The consequences of loss of cytoplasmic membrane anionic lipids, MreB, RodZ and/or YqiK suggest that the mode of action of the PspA effector is closely associated with cell envelope organization. © 2014 The Authors.

High throughput platforms for structural genomics of integral membrane proteins.

Science.gov (United States)

Mancia, Filippo; Love, James

2011-08-01

Structural genomics approaches on integral membrane proteins have been postulated for over a decade, yet specific efforts are lagging years behind their soluble counterparts. Indeed, high throughput methodologies for production and characterization of prokaryotic integral membrane proteins are only now emerging, while large-scale efforts for eukaryotic ones are still in their infancy. Presented here is a review of recent literature on actively ongoing structural genomics of membrane protein initiatives, with a focus on those aimed at implementing interesting techniques aimed at increasing our rate of success for this class of macromolecules. Copyright © 2011 Elsevier Ltd. All rights reserved.

Spin chain from membrane and the Neumann-Rosochatius integrable system

International Nuclear Information System (INIS)

Bozhilov, P.

2007-01-01

We find membrane configurations in AdS 4 xS 7 , which correspond to the continuous limit of the SU(2) integrable spin chain, considered as a limit of the SU(3) spin chain, arising in N=4 SYM in four dimensions, dual to strings in AdS 5 xS 5 . We also discuss the relationship with the Neumann-Rosochatius integrable system at the level of Lagrangians, comparing the string and membrane cases

The main thylakoid membrane lipid monogalactosyldiacylglycerol (MGDG) promotes the de-epoxidation of violaxanthin associated with the light-harvesting complex of photosystem II (LHCII).

Science.gov (United States)

Schaller, Susann; Latowski, Dariusz; Jemioła-Rzemińska, Małgorzata; Wilhelm, Christian; Strzałka, Kazimierz; Goss, Reimund

2010-03-01

In higher plants, the major part of the xanthophyll cycle pigment violaxanthin (Vx) is non-covalently bound to the main light-harvesting complex of PSII (LHCII). Under saturating light conditions Vx has to be released from its binding site into the surrounding lipid phase, where it is converted to zeaxanthin (Zx) by the enzyme Vx de-epoxidase (VDE). In the present study we investigated the influence of thylakoid lipids on the de-epoxidation of Vx, which was still associated with the LHCII. We isolated LHCII with different concentrations of native, endogenous lipids and Vx by sucrose gradient centrifugation or successive cation precipitation. Analysis of the different LHCII preparations showed that the concentration of LHCII-associated Vx was correlated with the concentration of the main thylakoid lipid monogalactosyldiacylglycerol (MGDG) associated with the complexes. Decreases in the MGDG content of the LHCII led to a diminished Vx concentration, indicating that a part of the total Vx pool was located in an MGDG phase surrounding the LHCII, whereas another part was bound to the LHCII apoproteins. We further studied the convertibility of LHCII-associated Vx in in-vitro enzyme assays by addition of isolated VDE. We observed an efficient and almost complete Vx conversion in the LHCII fractions containing high amounts of endogenous MGDG. LHCII preparations with low concentrations of MGDG exhibited a strongly reduced Vx de-epoxidation, which could be increased by addition of exogenous, pure MGDG. The de-epoxidation of LHCII-associated Vx was saturated at a much lower concentration of native, endogenous MGDG compared with the concentration of isolated, exogenous MGDG, which is needed for optimal VDE activity in in-vitro assays employing pure isolated Vx. Copyright 2009 Elsevier B.V. All rights reserved.

Recovery of real dye bath wastewater using integrated membrane process: considering water recovery, membrane fouling and reuse potential of membranes.

Science.gov (United States)

Balcik-Canbolat, Cigdem; Sengezer, Cisel; Sakar, Hacer; Karagunduz, Ahmet; Keskinler, Bulent

2017-11-01

It has been recognized by the whole world that textile industry which produce large amounts of wastewater with strong color and toxic organic compounds is a major problematical industry requiring effective treatment solutions. In this study, reverse osmosis (RO) membranes were tested on biologically treated real dye bath wastewater with and without pretreatment by nanofiltration (NF) membrane to recovery. Also membrane fouling and reuse potential of membranes were investigated by multiple filtrations. Obtained results showed that only NF is not suitable to produce enough quality to reuse the wastewater in a textile industry as process water while RO provide successfully enough permeate quality. The results recommend that integrated NF/RO membrane process is able to reduce membrane fouling and allow long-term operation for real dye bath wastewater.

HAMLET interacts with lipid membranes and perturbs their structure and integrity.

Science.gov (United States)

Mossberg, Ann-Kristin; Puchades, Maja; Halskau, Øyvind; Baumann, Anne; Lanekoff, Ingela; Chao, Yinxia; Martinez, Aurora; Svanborg, Catharina; Karlsson, Roger

2010-02-23

Cell membrane interactions rely on lipid bilayer constituents and molecules inserted within the membrane, including specific receptors. HAMLET (human alpha-lactalbumin made lethal to tumor cells) is a tumoricidal complex of partially unfolded alpha-lactalbumin (HLA) and oleic acid that is internalized by tumor cells, suggesting that interactions with the phospholipid bilayer and/or specific receptors may be essential for the tumoricidal effect. This study examined whether HAMLET interacts with artificial membranes and alters membrane structure. We show by surface plasmon resonance that HAMLET binds with high affinity to surface adherent, unilamellar vesicles of lipids with varying acyl chain composition and net charge. Fluorescence imaging revealed that HAMLET accumulates in membranes of vesicles and perturbs their structure, resulting in increased membrane fluidity. Furthermore, HAMLET disrupted membrane integrity at neutral pH and physiological conditions, as shown by fluorophore leakage experiments. These effects did not occur with either native HLA or a constitutively unfolded Cys-Ala HLA mutant (rHLA(all-Ala)). HAMLET also bound to plasma membrane vesicles formed from intact tumor cells, with accumulation in certain membrane areas, but the complex was not internalized by these vesicles or by the synthetic membrane vesicles. The results illustrate the difference in membrane affinity between the fatty acid bound and fatty acid free forms of partially unfolded HLA and suggest that HAMLET engages membranes by a mechanism requiring both the protein and the fatty acid. Furthermore, HAMLET binding alters the morphology of the membrane and compromises its integrity, suggesting that membrane perturbation could be an initial step in inducing cell death.

Ferredoxin-dependent and antimycin A-sensitive reduction of cytochrome b-559 by far-red light in maize [Zea mays] thylakoids; participation of a meladiol-reducible cytochrome b-559 in cyclic electron flow

International Nuclear Information System (INIS)

Miyake, C.; Schreiber, U.; Asada, K.

1995-01-01

Thylakoids from mesophyll cells of maize showed a high rate of the ferredoxin (Fd)-dependent and antimycin A (AntiA)-sensitive cyclic electron flow as determined by the quenching of 9-aminoacridine fluorescence which indicates the formation of ΔpH across the membranes. Spectrophotometric survey of the thylakoids showed the reduction of a Cyt having an α-peak at 559 nm [Cyt b-559(Fd)] by far-red light, which depended on Fd and was sensitive to AntiA. Dose dependencies of Fd and AntiA on the photoreduction of Cyt b-559(Fd) were the same as those of the formation of ΔpH. Cyt b-559(Fd) occurred in an oxidized form even in the presence of ascorbate and was reduced by far-red light. In darkness, it was reduced only by menadiol (E m,7 = –10mV). Thus, Cyt b-559(Fd) was distinguished from Cyt b-559 in the PSII complex by its low redox potential. The present results indicate that Cyt b-559(Fd) mediates electron transfer from Fd to plastoquinone during Fd-dependent cyclic electron flow around PSI. (author)

Light-induced modification of plant plasma membrane ion transport.

Science.gov (United States)

Marten, I; Deeken, R; Hedrich, R; Roelfsema, M R G

2010-09-01

Light is not only the driving force for electron and ion transport in the thylakoid membrane, but also regulates ion transport in various other membranes of plant cells. Light-dependent changes in ion transport at the plasma membrane and associated membrane potential changes have been studied intensively over the last century. These studies, with various species and cell types, revealed that apart from regulation by chloroplasts, plasma membrane transport can be controlled by phytochromes, phototropins or channel rhodopsins. In this review, we compare light-dependent plasma membrane responses of unicellular algae (Eremosphaera and Chlamydomonas), with those of a multicellular alga (Chara), liverworts (Conocephalum), mosses (Physcomitrella) and several angiosperm cell types. Light-dependent plasma membrane responses of Eremosphaera and Chara are characterised by the dominant role of K(+) channels during membrane potential changes. In most other species, the Ca(2+)-dependent activation of plasma membrane anion channels represents a general light-triggered event. Cell type-specific responses are likely to have evolved by modification of this general response or through the development of additional light-dependent signalling pathways. Future research to elucidate these light-activated signalling chains is likely to benefit from the recent identification of S-type anion channel genes and proteins capable of regulating these channels.

On controllability of an integrated bioreactor and periodically operated membrane separation process

DEFF Research Database (Denmark)

Prado Rubio, Oscar Andres; Jørgensen, Sten Bay; Jonsson, Gunnar Eigil

the influence of membrane fouling. Previously, the REED and fermentation processes have been modeled and investigated separately (Prado- Rubio et al., 2011a; Boonmee, 2003). Additionally, a simple quasi-sequential strategy for integrated process design and control structure development has been proposed (Prado...... to understand the controlled operation of the integrated process, it is convenient to use a model based approach supported by experimental evidence. Recently, an integrated bioreactor and electrically driven membrane separation process (Reverse Electro- Enhanced Dialysis - REED) has been proposed as a method...... at a certain lactate concentration level. Hence, productivity can be enhanced by the in situ lactate removal from the cultivation broth during pH controlled fermentation. This can be done by means of ion exchange membranes and electrical potential gradients. The novelty of the integrated process lies...

Performance of integrated bioelectrochemical membrane reactor: Energy recovery, pollutant removal and membrane fouling alleviation

Science.gov (United States)

Dong, Yue; He, Weihua; Li, Chao; Liang, Dandan; Qu, Youpeng; Han, Xiaoyu; Feng, Yujie

2018-04-01

A novel hybrid bioelectrochemical membrane reactor with integrated microfiltration membrane as the separator between electrodes is developed for domestic wastewater treatment. After accumulation of biofilm, the organic pollutants are mainly degraded in anodic compartment, and microfiltration membrane blocks the adverse leakage of dissolved oxygen from aerated cathodic compartment. The maximum system power output is restricted by gas-water ratio following a Monod-like relationship. Within the tested gas-water ratios ranging from 0.6 to 42.9, the half-saturation constant (KQ) is 5.9 ± 0.9 with a theoretic maximum power density of 20.4 ± 1.0 W m-3. Energy balance analysis indicates an appropriate gas-water ratio regulation (from 2.3 to 28.6) for cathodic compartment is necessary to obtain positive energy output for the system. A maximum net electricity output is 9.09 × 10-3 kWh m-3 with gas-water ratio of 17.1. Notably, the system achieves the chemical oxygen demand removal of 98.3 ± 0.3%, ammonia nitrogen removal of 99.6 ± 0.1%, and total nitrogen removal of 80.0 ± 0.9%. This work verifies an effective integration of microfiltration membrane into bioelectrochemical system as separator for high-quality effluent and provides an insight into the operation and regulation of biocathode system for effective electrical energy output.

High yield cell-free production of integral membrane proteins without refolding or detergents.

Science.gov (United States)

Wuu, Jessica J; Swartz, James R

2008-05-01

Integral membrane proteins act as critical cellular components and are important drug targets. However, difficulties in producing membrane proteins have hampered investigations of structure and function. In vivo production systems are often limited by cell toxicity, and previous in vitro approaches have required unnatural folding pathways using detergents or lipid solutions. To overcome these limitations, we present an improved cell-free expression system which produces high yields of integral membrane proteins without the use of detergents or refolding steps. Our cell-free reaction activates an Escherichia coli-derived cell extract for transcription and translation. Purified E. coli inner membrane vesicles supply membrane-bound components and the lipid environment required for insertion and folding. Using this system, we demonstrated successful synthesis of two complex integral membrane transporters, the tetracycline pump (TetA) and mannitol permease (MtlA), in yields of 570+/-50 microg/mL and 130+/-30 microg/mL of vesicle-associated protein, respectively. These yields are up to 400 times typical in vivo concentrations. Insertion and folding of these proteins are verified by sucrose flotation, protease digestion, and activity assays. Whereas TetA incorporates efficiently into vesicle membranes with over two-thirds of the synthesized protein being inserted, MtlA yields appear to be limited by insufficient concentrations of a membrane-associated chaperone.

Ethanol fermentation integrated with PDMS composite membrane: An effective process.

Science.gov (United States)

Fu, Chaohui; Cai, Di; Hu, Song; Miao, Qi; Wang, Yong; Qin, Peiyong; Wang, Zheng; Tan, Tianwei

2016-01-01

The polydimethylsiloxane (PDMS) membrane, prepared in water phase, was investigated in separation ethanol from model ethanol/water mixture and fermentation-pervaporation integrated process. Results showed that the PDMS membrane could effectively separate ethanol from model solution. When integrated with batch ethanol fermentation, the ethanol productivity was enhanced compared with conventional process. Fed-batch and continuous ethanol fermentation with pervaporation were also performed and studied. 396.2-663.7g/m(2)h and 332.4-548.1g/m(2)h of total flux with separation factor of 8.6-11.7 and 8-11.6, were generated in the fed-batch and continuous fermentation with pervaporation scenario, respectively. At the same time, high titre ethanol production of ∼417.2g/L and ∼446.3g/L were also achieved on the permeate side of membrane in the two scenarios, respectively. The integrated process was environmental friendly and energy saving, and has a promising perspective in long-terms operation. Copyright © 2015 Elsevier Ltd. All rights reserved.

High quality single crystal Ge nano-membranes for opto-electronic integrated circuitry

Energy Technology Data Exchange (ETDEWEB)

Shah, V. A., E-mail: vishal.shah@warwick.ac.uk; Gammon, P. M. [Department of Engineering, The University of Warwick, Coventry CV4 7AL (United Kingdom); Department of Physics, The University of Warwick, Coventry CV4 7AL (United Kingdom); Rhead, S. D.; Halpin, J. E.; Trushkevych, O.; Wilson, N. R.; Myronov, M.; Edwards, R. S.; Patchett, D. H.; Allred, P. S.; Prest, M. J.; Whall, T. E.; Parker, E. H. C.; Leadley, D. R. [Department of Physics, The University of Warwick, Coventry CV4 7AL (United Kingdom); Chávez-Ángel, E. [ICN2-Institut Catala de Nanociencia i Nanotecnologia, Campus UAB, 08193 Bellaterra (Barcelona) (Spain); Department of Physics, UAB, 08193 Bellaterra (Barcelona) (Spain); Shchepetov, A.; Prunnila, M. [VTT Technical Research Centre of Finland, P.O. Box 1000, FI-02044 VTT, Espoo (Finland); Kachkanov, V.; Dolbnya, I. P. [Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE (United Kingdom); Reparaz, J. S. [ICN2-Institut Catala de Nanociencia i Nanotecnologia, Campus UAB, 08193 Bellaterra (Barcelona) (Spain); and others

2014-04-14

A thin, flat, and single crystal germanium membrane would be an ideal platform on which to mount sensors or integrate photonic and electronic devices, using standard silicon processing technology. We present a fabrication technique compatible with integrated-circuit wafer scale processing to produce membranes of thickness between 60 nm and 800 nm, with large areas of up to 3.5 mm{sup 2}. We show how the optical properties change with thickness, including appearance of Fabry-Pérot type interference in thin membranes. The membranes have low Q-factors, which allow the platforms to counteract distortion during agitation and movement. Finally, we report on the physical characteristics showing sub-nm roughness and a homogenous strain profile throughout the freestanding layer, making the single crystal Ge membrane an excellent platform for further epitaxial growth or deposition of materials.

High quality single crystal Ge nano-membranes for opto-electronic integrated circuitry

International Nuclear Information System (INIS)

Shah, V. A.; Gammon, P. M.; Rhead, S. D.; Halpin, J. E.; Trushkevych, O.; Wilson, N. R.; Myronov, M.; Edwards, R. S.; Patchett, D. H.; Allred, P. S.; Prest, M. J.; Whall, T. E.; Parker, E. H. C.; Leadley, D. R.; Chávez-Ángel, E.; Shchepetov, A.; Prunnila, M.; Kachkanov, V.; Dolbnya, I. P.; Reparaz, J. S.

2014-01-01

A thin, flat, and single crystal germanium membrane would be an ideal platform on which to mount sensors or integrate photonic and electronic devices, using standard silicon processing technology. We present a fabrication technique compatible with integrated-circuit wafer scale processing to produce membranes of thickness between 60 nm and 800 nm, with large areas of up to 3.5 mm 2 . We show how the optical properties change with thickness, including appearance of Fabry-Pérot type interference in thin membranes. The membranes have low Q-factors, which allow the platforms to counteract distortion during agitation and movement. Finally, we report on the physical characteristics showing sub-nm roughness and a homogenous strain profile throughout the freestanding layer, making the single crystal Ge membrane an excellent platform for further epitaxial growth or deposition of materials

Spectral studies of Lanthanide interactions with membrane surfaces

Energy Technology Data Exchange (ETDEWEB)

Karukstis, K.K.; Kao, M.Y.; Savin, D.A.; Bittker, R.A.; Kaphengst, K.J.; Emetarom, C.M.; Naito, N.R.; Takamoto, D.Y. [Harvey Mudd College, Claremont, CA (United States)

1995-03-23

We have monitored the interactions of the series of trivalent lanthanide cations with the thylakoid membrane surface of spinach chloroplasts using two complementary spectral techniques. Measurements of the fluorescence emission of the extrinsic probe 2-p-toluidinonaphthalene-6-sulfonate (TNS) and the absorbance of the intrinsic chromophore chlorophyll provide two sensitive means of characterizing the dependence of the cation-membrane interaction on the nature of the cation. In these systems, added lanthanide cations adsorb onto the membrane surface to neutralize exposed segments of membrane-embedded protein complexes. The lanthanide-induced charge neutralization increases the proximity of added TNS anion to the membrane surface as evidenced by variations in the TNS fluorescence level and wavelength of maximum emission. Our results reveal a strong dependence of TNS fluorescence parameters on both lanthanide size and total orbital angular momentum L value. Lanthanides with greater charge density (small size and/or low L value) enhance the TNS fluorescence level to a greater extent. A possible origin for the lanthanide-dependent TNS fluorescence levels is suggested in terms of a heterogeneity in the number and type of TNS binding sites. The data are consistent with the proposal that larger lanthanides with smaller enthalpies of hydration induce more significant membrane appression. 59 refs., 9 figs., 2 tabs.

Quality control of Photosystem II: reversible and irreversible protein aggregation decides the fate of Photosystem II under excessive illumination

Directory of Open Access Journals (Sweden)

Yasusi eYamamoto

2013-10-01

Full Text Available In response to excessive light, the thylakoid membranes of higher plant chloroplasts show dynamic changes including the degradation and reassembly of proteins, a change in the distribution of proteins, and large-scale structural changes such as unstacking of the grana. Here, we examined the aggregation of light-harvesting chlorophyll-protein complexes and Photosystem II core subunits of spinach thylakoid membranes under light stress with 77K chlorophyll fluorescence; aggregation of these proteins was found to proceed with increasing light intensity. Measurement of changes in the fluidity of thylakoid membranes with fluorescence polarization of diphenylhexatriene showed that membrane fluidity increased at a light intensity of 500–1,000 µmol photons m-2 s-1, and decreased at very high light intensity (1,500 µmol photons m-2 s-1. The aggregation of light-harvesting complexes at moderately high light intensity is known to be reversible, while that of Photosystem II core subunits at extremely high light intensity is irreversible. It is likely that the reversibility of protein aggregation is closely related to membrane fluidity: increases in fluidity should stimulate reversible protein aggregation, whereas irreversible protein aggregation might decrease membrane fluidity. When spinach leaves were pre-illuminated with moderately high light intensity, the qE component of non-photochemical quenching and the optimum quantum yield of Photosystem II increased, indicating that Photosystem II/ light-harvesting complexes rearranged in the thylakoid membranes to optimize Photosystem II activity. Transmission electron microscopy revealed that the thylakoids underwent partial unstacking under these light stress conditions. Thus, protein aggregation is involved in thylakoid dynamics and regulates photochemical reactions, thereby deciding the fate of Photosystem II.

Polyethylenimine-mediated impairment of mitochondrial membrane potential, respiration and membrane integrity

DEFF Research Database (Denmark)

Larsen, Anna Karina; Malinska, Dominika; Koszela-Piotrowska, Izabela

2012-01-01

The 25 kDa branched polyethylenimine (PEI) is a highly efficient synthetic polycation used in transfection protocols, but also triggers mitochondrial-mediated apoptotic cell death processes where the mechanistic issues are poorly understood. We now demonstrate that PEI in a concentration- and time......-dependent manner can affect functions (membrane potential, swelling and respiration) and ultrastructural integrity of freshly isolated rat liver mitochondria. The threshold concentration for detection of PEI-mediated impairment of rat liver mitochondrial functions is 3 µg/mL, however, lower PEI levels still exert...... some effects on mitochondrial morphology and respiration, and these may be related to the inherent membrane perturbing properties of this polycation. The PEI-mediated mitochondrial swelling phase is biphasic, with a fast decaying initial period (most prominent from 4 µg/mL PEI) followed by a slower...

Phenotype abnormality: 47 [Arabidopsis Phenome Database[Archive

Lifescience Database Archive (English)

Full Text Available 47 http://metadb.riken.jp/db/SciNetS_ria224i/cria224u1ria224u553i abnormal for trait of behavior...al quality in organ named thylakoid membrane during process named thylakoid membrane organization ... abnormal ... behavioral quality

Association of lipids with integral membrane surface proteins of Mycoplasma hyorhinis

International Nuclear Information System (INIS)

Bricker, T.M.; Boyer, M.J.; Keith, J.; Watson-McKown, R.; Wise, K.S.

1988-01-01

Triton X-114 (TX-114)-phase fractionation was used to identify and characterize integral membrane surface proteins of the wall-less procaryote Mycoplasma hyorhinis GDL. Phase fractionation of mycoplasmas followed by analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed selective partitioning of approximately 30 [ 35 S]methionine-labeled intrinsic membrane proteins into the TX-114 phase. Similar analysis of [ 3 H]palmitate-labeled cells showed that approximately 20 proteins of this organism were associated with lipid, all of which also efficiently partitioned as integral membrane components into the detergent phase. Immunoblotting and immunoprecipitation of TX-114-phase proteins from 125 I-surface-labeled cells with four monoclonal antibodies to distinct surface epitopes of M. hyorhinis identified surface proteins p120, p70, p42, and p23 as intrinsic membrane components. Immunoprecipitation of [ 3 H]palmitate-labeled TX-114-phase proteins further established that surface proteins p120, p70, and p23 (a molecule that mediates complement-dependent mycoplasmacidal monoclonal antibody activity) were among the lipid-associated proteins of this organism. Two of these proteins, p120 and p123, were acidic (pI less than or equal to 4.5), as shown by two-dimensional isoelectric focusing. This study established that M. hyorhinis contains an abundance of integral membrane proteins tightly associated with lipids and that many of these proteins are exposed at the external surface of the single limiting plasma membrane. Monoclonal antibodies are reported that will allow detailed analysis of the structure and processing of lipid-associated mycoplasma proteins

Transfer Printed Nanomembranes for Heterogeneously Integrated Membrane Photonics

Directory of Open Access Journals (Sweden)

Hongjun Yang

2015-11-01

Full Text Available Heterogeneous crystalline semiconductor nanomembrane (NM integration is investigated for single-layer and double-layer Silicon (Si NM photonics, III-V/Si NM lasers, and graphene/Si NM total absorption devices. Both homogeneous and heterogeneous integration are realized by the versatile transfer printing technique. The performance of these integrated membrane devices shows, not only intact optical and electrical characteristics as their bulk counterparts, but also the unique light and matter interactions, such as Fano resonance, slow light, and critical coupling in photonic crystal cavities. Such a heterogeneous integration approach offers tremendous practical application potentials on unconventional, Si CMOS compatible, and high performance optoelectronic systems.

Integral membrane protein structure determination using pseudocontact shifts

Energy Technology Data Exchange (ETDEWEB)

Crick, Duncan J.; Wang, Jue X. [University of Cambridge, Department of Biochemistry (United Kingdom); Graham, Bim; Swarbrick, James D. [Monash University, Monash Institute of Pharmaceutical Sciences (Australia); Mott, Helen R.; Nietlispach, Daniel, E-mail: dn206@cam.ac.uk [University of Cambridge, Department of Biochemistry (United Kingdom)

2015-04-15

Obtaining enough experimental restraints can be a limiting factor in the NMR structure determination of larger proteins. This is particularly the case for large assemblies such as membrane proteins that have been solubilized in a membrane-mimicking environment. Whilst in such cases extensive deuteration strategies are regularly utilised with the aim to improve the spectral quality, these schemes often limit the number of NOEs obtainable, making complementary strategies highly beneficial for successful structure elucidation. Recently, lanthanide-induced pseudocontact shifts (PCSs) have been established as a structural tool for globular proteins. Here, we demonstrate that a PCS-based approach can be successfully applied for the structure determination of integral membrane proteins. Using the 7TM α-helical microbial receptor pSRII, we show that PCS-derived restraints from lanthanide binding tags attached to four different positions of the protein facilitate the backbone structure determination when combined with a limited set of NOEs. In contrast, the same set of NOEs fails to determine the correct 3D fold. The latter situation is frequently encountered in polytopical α-helical membrane proteins and a PCS approach is thus suitable even for this particularly challenging class of membrane proteins. The ease of measuring PCSs makes this an attractive route for structure determination of large membrane proteins in general.

The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants.

Science.gov (United States)

Tyutereva, Elena V; Evkaikina, Anastasiia I; Ivanova, Alexandra N; Voitsekhovskaja, Olga V

2017-09-01

The lateral mobility of integral components of thylakoid membranes, such as plastoquinone, xanthophylls, and pigment-protein complexes, is critical for the maintenance of efficient light harvesting, high rates of linear electron transport, and successful repair of damaged photosystem II (PSII). The packaging of the photosynthetic pigment-protein complexes in the membrane depends on their size and stereometric parameters which in turn depend on the composition of the complexes. Chlorophyll b (Chlb) is an important regulator of antenna size and composition. In this study, the lateral mobility (the mobile fraction size) of pigment-protein complexes and lipids in grana membranes was analyzed in chlorina mutants of Arabidopsis and barley lacking Chlb. In the Arabidopsis ch1-3 mutant, diffusion of membrane lipids decreased as compared to wild-type plants, but the diffusion of photosynthetic complexes was not affected. In the barley chlorina f2 3613 mutant, the diffusion of pigment-protein complexes significantly decreased, while the diffusion of lipids increased, as compared to wild-type plants. We propose that the size of the mobile fractions of pigment-protein complexes in grana membranes in vivo is higher than reported previously. The data are discussed in the context of the protein composition of antennae, characteristics of the plastoquinone pool, and production of reactive oxygen species in leaves of chlorina mutants.

Changes in photosynthesis and activities of enzymes involved in ...

African Journals Online (AJOL)

AJL

2012-04-26

Apr 26, 2012 ... oxygen and carbohydrates. In photosynthesis, a series of redox reactions occur in the electron transport system present in the chloroplast thylakoid membranes. Oxi- dation of water is catalyzed by photosystem II (PSII), a multi-subunit pigment protein complex located in the thylakoid membrane (Hillier and ...

Efficient ethanol recovery from yeast fermentation broth with integrated distillation-membrane process

Science.gov (United States)

A hybrid process integrating vapor stripping with vapor compression and vapor permeation membrane separation, termed Membrane Assisted Vapor Stripping (MAVS), was evaluated for recovery and dehydration of ethanol from aqueous solution as an alternative to conventional distillatio...

Proteomics of the chloroplast: systematic identification and targeting analysis of lumenal and peripheral thylakoid proteins

DEFF Research Database (Denmark)

Peltier, J B; Friso, G; Kalume, D E

2000-01-01

The soluble and peripheral proteins in the thylakoids of pea were systematically analyzed by using two-dimensional electrophoresis, mass spectrometry, and N-terminal Edman sequencing, followed by database searching. After correcting to eliminate possible isoforms and post-translational modificati......The soluble and peripheral proteins in the thylakoids of pea were systematically analyzed by using two-dimensional electrophoresis, mass spectrometry, and N-terminal Edman sequencing, followed by database searching. After correcting to eliminate possible isoforms and post......-translational modifications, we estimated that there are at least 200 to 230 different lumenal and peripheral proteins. Sixty-one proteins were identified; for 33 of these proteins, a clear function or functional domain could be identified, whereas for 10 proteins, no function could be assigned. For 18 proteins, no expressed...... sequence tag or full-length gene could be identified in the databases, despite experimental determination of a significant amount of amino acid sequence. Nine previously unidentified proteins with lumenal transit peptides are presented along with their full-length genes; seven of these proteins possess...

Thermoelectric integrated membrane evaporation water recovery technology

Science.gov (United States)

Roebelen, G. J., Jr.; Winkler, H. E.; Dehner, G. F.

1982-01-01

The recently developed Thermoelectric Integrated Membrane Evaporation Subsystem (TIMES) offers a highly competitive approach to water recovery from waste fluids for future on-orbit stations such as the Space Operations Center. Low power, compactness and gravity insensitive operation are featured in this vacuum distillation subsystem that combines a hollow fiber membrane evaporator with a thermoelectric heat pump. The hollow fiber elements provide positive liquid/gas phase control with no moving parts other than pumps and an accumulator, thus solving problems inherent in other reclamation subsystem designs. In an extensive test program, over 850 hours of operation were accumulated during which time high quality product water was recovered from both urine and wash water at an average steady state production rate of 2.2 pounds per hour.

The lysosomal membrane protein SCAV-3 maintains lysosome integrity and adult longevity

Science.gov (United States)

Li, Yuan; Chen, Baohui; Zou, Wei; Wang, Xin; Wu, Yanwei; Zhao, Dongfeng; Sun, Yanan; Liu, Yubing

2016-01-01

Lysosomes degrade macromolecules and recycle metabolites as well as being involved in diverse processes that regulate cellular homeostasis. The lysosome is limited by a single phospholipid bilayer that forms a barrier to separate the potent luminal hydrolases from other cellular constituents, thus protecting the latter from unwanted degradation. The mechanisms that maintain lysosomal membrane integrity remain unknown. Here, we identified SCAV-3, the Caenorhabditis elegans homologue of human LIMP-2, as a key regulator of lysosome integrity, motility, and dynamics. Loss of scav-3 caused rupture of lysosome membranes and significantly shortened lifespan. Both of these phenotypes were suppressed by reinforced expression of LMP-1 or LMP-2, the C. elegans LAMPs, indicating that longevity requires maintenance of lysosome integrity. Remarkably, reduction in insulin/insulin-like growth factor 1 (IGF-1) signaling suppressed lysosomal damage and extended the lifespan in scav-3(lf) animals in a DAF-16–dependent manner. Our data reveal that SCAV-3 is essential for preserving lysosomal membrane stability and that modulation of lysosome integrity by the insulin/IGF-1 signaling pathway affects longevity. PMID:27810910

Activity, polypeptide and gene identification of thylakoid Ndh complex in trees: potential physiological relevance of fluorescence assays.

Science.gov (United States)

Serrot, Patricia H; Sabater, Bartolomé; Martín, Mercedes

2012-09-01

Three evergreen (Laurus nobilis, Viburnum tinus and Thuja plicata) and two autumnal abscission deciduous trees (Cydonia oblonga and Prunus domestica) have been investigated for the presence (zymogram and immunodetection) and functionality (post-illumination chlorophyll fluorescence) of the thylakoid Ndh complex. The presence of encoding ndh genes has also been investigated in T. plicata. Western assays allowed tentative identification of zymogram NADH dehydrogenase bands corresponding to the Ndh complex after native electrophoresis of solubilized fractions from L. nobilis, V. tinus, C. oblonga and P. domestica leaves, but not in those of T. plicata. However, Ndh subunits were detected after SDS-PAGE of thylakoid solubilized proteins of T. plicata. The leaves of the five plants showed the post-illumination chlorophyll fluorescence increase dependent on the presence of active Ndh complex. The fluorescence increase was higher in autumn in deciduous, but not in evergreen trees, which suggests that the thylakoid Ndh complex could be involved in autumnal leaf senescence. Two ndhB genes were sequenced from T. plicata that differ at the 350 bp 3' end sequence. Comparison with the mRNA revealed that ndhB genes have a 707-bp type II intron between exons 1 (723 bp) and 2 (729 bp) and that the UCA 259th codon is edited to UUA in mRNA. Phylogenetically, the ndhB genes of T. plicata group close to those of Metasequoia, Cryptomeria, Taxodium, Juniperus and Widdringtonia in the cupresaceae branch and are 5' end shortened by 18 codons with respect to that of angiosperms. Copyright © Physiologia Plantarum 2012.

Integrated approach to characterize fouling on a flat sheet membrane gravity driven submerged membrane bioreactor

KAUST Repository

Fortunato, Luca

2016-10-07

Fouling in membrane bioreactors (MBR) is acknowledged to be complex and unclear. An integrated characterization methodology was employed in this study to understand the fouling on a gravity-driven submerged MBR (GD-SMBR). It involved the use of different analytical tools, including optical coherence tomography (OCT), liquid chromatography with organic carbon detection (LC-OCD), total organic carbon (TOC), flow cytometer (FCM), adenosine triphosphate analysis (ATP) and scanning electron microscopy (SEM). The three-dimensional (3D) biomass morphology was acquired in a real-time through non-destructive and in situ OCT scanning of 75% of the total membrane surface directly in the tank. Results showed that the biomass layer was homogeneously distributed on the membrane surface. The amount of biomass was selectively linked with final destructive autopsy techniques. The LC-OCD analysis indicated the abundance of low molecular weight (LMW) organics in the fouling composition. Three different SEM techniques were applied to investigate the detailed fouling morphology on the membrane. © 2016 Elsevier Ltd

Integrated approach to characterize fouling on a flat sheet membrane gravity driven submerged membrane bioreactor.

Science.gov (United States)

Fortunato, Luca; Jeong, Sanghyun; Wang, Yiran; Behzad, Ali R; Leiknes, TorOve

2016-12-01

Fouling in membrane bioreactors (MBR) is acknowledged to be complex and unclear. An integrated characterization methodology was employed in this study to understand the fouling on a gravity-driven submerged MBR (GD-SMBR). It involved the use of different analytical tools, including optical coherence tomography (OCT), liquid chromatography with organic carbon detection (LC-OCD), total organic carbon (TOC), flow cytometer (FCM), adenosine triphosphate analysis (ATP) and scanning electron microscopy (SEM). The three-dimensional (3D) biomass morphology was acquired in a real-time through non-destructive and in situ OCT scanning of 75% of the total membrane surface directly in the tank. Results showed that the biomass layer was homogeneously distributed on the membrane surface. The amount of biomass was selectively linked with final destructive autopsy techniques. The LC-OCD analysis indicated the abundance of low molecular weight (LMW) organics in the fouling composition. Three different SEM techniques were applied to investigate the detailed fouling morphology on the membrane. Copyright © 2016 Elsevier Ltd. All rights reserved.

The effectiveness of styrene-maleic acid (SMA) copolymers for solubilisation of integral membrane proteins from SMA-accessible and SMA-resistant membranes.

Science.gov (United States)

Swainsbury, David J K; Scheidelaar, Stefan; Foster, Nicholas; van Grondelle, Rienk; Killian, J Antoinette; Jones, Michael R

2017-10-01

Solubilisation of biological lipid bilayer membranes for analysis of their protein complement has traditionally been carried out using detergents, but there is increasing interest in the use of amphiphilic copolymers such as styrene maleic acid (SMA) for the solubilisation, purification and characterisation of integral membrane proteins in the form of protein/lipid nanodiscs. Here we survey the effectiveness of various commercially-available formulations of the SMA copolymer in solubilising Rhodobacter sphaeroides reaction centres (RCs) from photosynthetic membranes. We find that formulations of SMA with a 2:1 or 3:1 ratio of styrene to maleic acid are almost as effective as detergent in solubilising RCs, with the best solubilisation by short chain variants (membranes was uniformly low, but could be increased through a variety of treatments to increase the lipid:protein ratio. However, proteins isolated from such membranes comprised clusters of complexes in small membrane patches rather than individual proteins. We conclude that short-chain 2:1 and 3:1 formulations of SMA are the most effective in solubilising integral membrane proteins, but that solubilisation efficiencies are strongly influenced by the size of the target protein and the density of packing of proteins in the membrane. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

Studies on improved integrated membrane-based chromatographic process for bioseparation

Science.gov (United States)

Xu, Yanke

To improve protein separation and purification directly from a fermentation broth, a novel membrane filtration-cum-chromatography device configuration having a relatively impermeable coated zone near the hollow fiber module outlet has been developed. The integrated membrane filtration-cum-chromatography unit packed with chromatographic beads on the shell side of the hollow fiber unit enjoys the advantages of both membrane filtration and chromatography; it allows one to load the chromatographic media directly from the fermentation broth or lysate and separate the adsorbed proteins through the subsequent elution step in a cyclic process. Interfacial polymerization was carried out to coat the bottom section of the hollow fiber membrane while leaving the rest of the hollow fiber membrane unaffected. Myoglobin (Mb), bovine serum albumin (BSA) and a-lactalbumin (a-LA) were used as model proteins in binary mixtures. Separation behaviors of binary protein mixtures were studied in devices using either an ultrafiltration (UF) membrane or a microfiltration (MF) membrane. Experimental results show that the breakthrough time and the protein loading capacities were dramatically improved after coating in both UF and MF modules. For a synthetic yeast fermentation broth feed, the Mb and a-LA elution profiles for the four consecutive cyclic runs were almost superimposable. Due to the lower transmembrane flux in this device plus the periodical washing-elution during the chromatographic separation, fouling was not a problem as it is in conventional microfiltration. A mathematical model describing the hydrodynamic and protein loading behaviors of the integrated device using UF membrane with a coated zone was developed. The simulation results for the breakthrough agree well with the experimental breakthrough curves. The optimal length of the coated zone was obtained from the simulation. A theoretical analysis of the protein mass transfer was performed using a diffusion-convection model

Development of a preprototype thermoelectric integrated membrane evaporation subsystem for water recovery

Science.gov (United States)

Winkler, H. E.; Roebelen, G. J., Jr.

1980-01-01

A three-man urine water recovery preprototype subsystem using a new concept to provide efficient potable water recovery from waste fluids on extended duration space flights has been designed, fabricated, and tested. Low power, compactness, and gravity insensitive operation are featured in this vacuum distillation subsystem that combines a hollow fiber polysulfone membrane evaporator with a thermoelectric heat pump. Application and integration of these key elements have solved problems inherent in previous reclamation subsystem designs. The hollow fiber elements provide positive liquid/gas phase control with no moving parts other than a waste liquid recirculation pump and a product water withdrawal pump. Tubular membranes provide structural integrity, improving on previous flat sheet membrane designs. A thermoelectric heat pump provides latent energy recovery.

Distinguishing the Roles of Thylakoid Respiratory Terminal Oxidases in the Cyanobacterium Synechocystis sp. PCC 6803.

Science.gov (United States)

Ermakova, Maria; Huokko, Tuomas; Richaud, Pierre; Bersanini, Luca; Howe, Christopher J; Lea-Smith, David J; Peltier, Gilles; Allahverdiyeva, Yagut

2016-06-01

Various oxygen-utilizing electron sinks, including the soluble flavodiiron proteins (Flv1/3), and the membrane-localized respiratory terminal oxidases (RTOs), cytochrome c oxidase (Cox) and cytochrome bd quinol oxidase (Cyd), are present in the photosynthetic electron transfer chain of Synechocystis sp. PCC 6803. However, the role of individual RTOs and their relative importance compared with other electron sinks are poorly understood, particularly under light. Via membrane inlet mass spectrometry gas exchange, chlorophyll a fluorescence, P700 analysis, and inhibitor treatment of the wild type and various mutants deficient in RTOs, Flv1/3, and photosystem I, we investigated the contribution of these complexes to the alleviation of excess electrons in the photosynthetic chain. To our knowledge, for the first time, we demonstrated the activity of Cyd in oxygen uptake under light, although it was detected only upon inhibition of electron transfer at the cytochrome b6f site and in ∆flv1/3 under fluctuating light conditions, where linear electron transfer was drastically inhibited due to impaired photosystem I activity. Cox is mostly responsible for dark respiration and competes with P700 for electrons under high light. Only the ∆cox/cyd double mutant, but not single mutants, demonstrated a highly reduced plastoquinone pool in darkness and impaired gross oxygen evolution under light, indicating that thylakoid-based RTOs are able to compensate partially for each other. Thus, both electron sinks contribute to the alleviation of excess electrons under illumination: RTOs continue to function under light, operating on slower time ranges and on a limited scale, whereas Flv1/3 responds rapidly as a light-induced component and has greater capacity. © 2016 American Society of Plant Biologists. All Rights Reserved.

Integrated Structural Biology for α-Helical Membrane Protein Structure Determination.

Science.gov (United States)

Xia, Yan; Fischer, Axel W; Teixeira, Pedro; Weiner, Brian; Meiler, Jens

2018-04-03

While great progress has been made, only 10% of the nearly 1,000 integral, α-helical, multi-span membrane protein families are represented by at least one experimentally determined structure in the PDB. Previously, we developed the algorithm BCL::MP-Fold, which samples the large conformational space of membrane proteins de novo by assembling predicted secondary structure elements guided by knowledge-based potentials. Here, we present a case study of rhodopsin fold determination by integrating sparse and/or low-resolution restraints from multiple experimental techniques including electron microscopy, electron paramagnetic resonance spectroscopy, and nuclear magnetic resonance spectroscopy. Simultaneous incorporation of orthogonal experimental restraints not only significantly improved the sampling accuracy but also allowed identification of the correct fold, which is demonstrated by a protein size-normalized transmembrane root-mean-square deviation as low as 1.2 Å. The protocol developed in this case study can be used for the determination of unknown membrane protein folds when limited experimental restraints are available. Copyright © 2018 Elsevier Ltd. All rights reserved.

Effects of Bloom-Forming Algae on Fouling of Integrated Membrane Systems in Seawater Desalination

Science.gov (United States)

Ladner, David Allen

2009-01-01

Combining low- and high-pressure membranes into an integrated membrane system is an effective treatment strategy for seawater desalination. Low-pressure microfiltration (MF) and ultrafiltration (UF) membranes remove particulate material, colloids, and high-molecular-weight organics leaving a relatively foulant-free salt solution for treatment by…

Trienoic fatty acids and plant tolerance of temperature

Directory of Open Access Journals (Sweden)

Routaboul Jean-Marc

2002-01-01

Full Text Available The biophysical reactions of light harvesting and electron transport during photosynthesis take place in a uniquely constructed bilayer, the thylakoid. In all photosynthetic eukaryotes, the complement of atypical glycerolipid molecules that form the foundation of this membrane are characterised by sugar head-groups and a very high level of unsaturation in the fatty acids that occupy the central portion of the thylakoid bilayer. alpha-linolenic (18:3 or a combination of 18:3 and hexadecatrienoic (16:3 acids typically account for approximately two-thirds of all thylakoid membrane fatty acids and over 90% of the fatty acids of monogalactosyl diacylglycerol, the major thylakoid lipid [1, 2]. The occurrence of trienoic fatty acids as a major component of the thylakoid membrane is especially remarkable since these fatty acids form highly reactive targets for active oxygen species and free radicals, which are often the by-products of oxygenic photosynthesis. Photosynthesis is one of the most temperature-sensitive functions of plant [3, 4]. There remains a widespread belief that these trienoic fatty acids might have some crucial role in plants to be of such universal occurrence, especially in photosynthesis tolerance of temperature [5].

Integrated investigation approach for determining mechanical properties of poly-silicon membranes

OpenAIRE

Brueckner, J.; Dehe, A.; Auerswald, E.; Dudek, R.; Michel, B.; Rzepka, S.

2014-01-01

A methodology is presented for determining mechanical properties of free-standing thin films such as poly-silicon membranes. The integrated investigation approach comprises test structure development, mechanical testing, and numerical simulation. All membrane test structures developed and manufactured consist of the same material but have different stiffness due to variations in the geometric design. The mechanical tests apply microscopic loads utilizing a nanoindentation tool. Young's modulu...

Integrable systems from membranes on AdS4 x S7

International Nuclear Information System (INIS)

Bozhilov, P.

2008-01-01

We describe how Neumann and Neumann-Rosochatius type integrable systems, as well as the continuous limit of the SU(2) integrable spin chain, can be obtained from membranes on AdS 4 x S 7 background, in the framework of AdS/CFT correspondence. (Abstract Copyright [2008], Wiley Periodicals, Inc.)

Membrane skeletal proteins and their integral membrane protein anchors are targets for tyrosine and threonine kinases in Euglena.

Science.gov (United States)

Fazio, M J; Da Silva, A C; Rosiere, T K; Bouck, G B

1995-01-01

Proteins of the membrane skeleton of Euglena gracilis were extensively phosphorylated in vivo and in vitro after incubation with [32P]-orthophosphate or gamma-[32P] ATP. Endogenous protein threonine/serine activity phosphorylated the major membrane skeletal proteins (articulins) and the putative integral membrane protein (IP39) anchor for articulins. The latter was also the major target for endogenous protein tyrosine kinase activity. A cytoplasmic domain of IP39 was specifically phosphorylated, and removal of this domain with papain eliminated the radiolabeled phosphoamino acids and eliminated or radically shifted the PI of the multiple isoforms of IP39. In gel kinase assays IP39 autophosphorylated and a 25 kDa protein which does not autophosphorylate was identified as a threonine/serine (casein) kinase. Plasma membranes from the membrane skeletal protein complex contained threonine/serine (casein) kinase activity, and cross-linking experiments suggested that IP39 was the likely source for this membrane activity. pH optima, cation requirements and heparin sensitivity of the detergent solubilized membrane activity were determined. Together these results suggest that protein kinases may be important modulators of protein assembly and function of the membrane skeleton of these protistan cells.

Investigating the role of viral integral membrane proteins in promoting the assembly of nepovirus and comovirus replication factories

Directory of Open Access Journals (Sweden)

Helene eSanfacon

2013-01-01

Full Text Available Formation of plant virus membrane-associated replication factories requires the association of viral replication proteins and viral RNA with intracellular membranes, the recruitment of host factors and the modification of membranes to form novel structures that house the replication complex. Many viruses encode integral membrane proteins that act as anchors for the replication complex. These hydrophobic proteins contain trans-membrane domains and/or amphipathic helices that associate with the membrane and modify its structure. The comovirus Co-Pro and NTP-binding (NTB, putative helicase proteins and the cognate nepovirus X2 and NTB proteins are among the best characterized plant virus integral membrane replication proteins and are functionally related to the picornavirus 2B, 2C and 3A membrane proteins. The identification of membrane-association domains and analysis of the membrane topology of these proteins is discussed. The evidence suggesting that these proteins have the ability to induce membrane proliferation, alter the structure and integrity of intracellular membranes and modulate the induction of symptoms in infected plants is also reviewed. Finally, areas of research that need further investigation are highlighted.

α-Synuclein oligomers induced by docosahexaenoic acid affect membrane integrity.

Directory of Open Access Journals (Sweden)

Chiara Fecchio

Full Text Available A key feature of Parkinson disease is the aggregation of α-synuclein and its intracellular deposition in fibrillar form. Increasing evidence suggests that the pathogenicity of α-synuclein is correlated with the activity of oligomers formed in the early stages of its aggregation process. Oligomers toxicity seems to be associated with both their ability to bind and affect the integrity of lipid membranes. Previously, we demonstrated that α-synuclein forms oligomeric species in the presence of docosahexaenoic acid and that these species are toxic to cells. Here we studied how interaction of these oligomers with membranes results in cell toxicity, using cellular membrane-mimetic and cell model systems. We found that α-synuclein oligomers are able to interact with large and small unilamellar negatively charged vesicles acquiring an increased amount of α-helical structure, which induces small molecules release. We explored the possibility that oligomers effects on membranes could be due to pore formation, to a detergent-like effect or to fibril growth on the membrane. Our biophysical and cellular findings are consistent with a model where α-synuclein oligomers are embedded into the lipid bilayer causing transient alteration of membrane permeability.

First integrals of the axisymmetric shape equation of lipid membranes

Science.gov (United States)

Zhang, Yi-Heng; McDargh, Zachary; Tu, Zhan-Chun

2018-03-01

The shape equation of lipid membranes is a fourth-order partial differential equation. Under the axisymmetric condition, this equation was transformed into a second-order ordinary differential equation (ODE) by Zheng and Liu (Phys. Rev. E 48 2856 (1993)). Here we try to further reduce this second-order ODE to a first-order ODE. First, we invert the usual process of variational calculus, that is, we construct a Lagrangian for which the ODE is the corresponding Euler–Lagrange equation. Then, we seek symmetries of this Lagrangian according to the Noether theorem. Under a certain restriction on Lie groups of the shape equation, we find that the first integral only exists when the shape equation is identical to the Willmore equation, in which case the symmetry leading to the first integral is scale invariance. We also obtain the mechanical interpretation of the first integral by using the membrane stress tensor. Project supported by the National Natural Science Foundation of China (Grant No. 11274046) and the National Science Foundation of the United States (Grant No. 1515007).

INTRASURGICAL MICROSCOPE-INTEGRATED SPECTRAL DOMAIN OPTICAL COHERENCE TOMOGRAPHY-ASSISTED MEMBRANE PEELING.

Science.gov (United States)

Falkner-Radler, Christiane I; Glittenberg, Carl; Gabriel, Max; Binder, Susanne

2015-10-01

To evaluate microscope-integrated intrasurgical spectral domain optical coherence tomography during macular surgery in a prospective monocenter study. Before pars plana vitrectomy and before, during, and after membrane peeling, 512 × 128 macular cube scans were performed using a Carl Zeiss Meditec Cirrus high-definition OCT system adapted to the optical pathway of a Zeiss OPMI VISU 200 surgical microscope and compared with retinal staining. The study included 51 patients with epiretinal membranes, with 8 of those having additional lamellar macular holes, 11 patients with vitreomacular traction, and 8 patients with full-thickness macular holes. Intraoperative spectral domain optical coherence tomography allowed performing membrane peeling without using retinal dyes in 40% of cases (28 of 70 patients). No residual membranes were found in 94.3% of patients (66 of 70 patients) in intrasurgical spectral domain optical coherence tomography and subsequent (re)staining. In patients with vitreomacular traction, intrasurgical spectral domain optical coherence tomography scans facilitated decisions on the need for an intraocular tamponade after membrane peeling. Intraoperative spectral domain optical coherence tomography was comparable with retinal dyes in confirming success after membrane peeling. However, the visualization of flat membranes was better after staining.

Anchoring a plant cytochrome P450 via PsaM to the thylakoids in Synechococcus sp. PCC 7002: evidence for light-driven biosynthesis.

Directory of Open Access Journals (Sweden)

Lærke Münter Lassen

Full Text Available Plants produce an immense variety of specialized metabolites, many of which are of high value as their bioactive properties make them useful as for instance pharmaceuticals. The compounds are often produced at low levels in the plant, and due to their complex structures, chemical synthesis may not be feasible. Here, we take advantage of the reducing equivalents generated in photosynthesis in developing an approach for producing plant bioactive natural compounds in a photosynthetic microorganism by functionally coupling a biosynthetic enzyme to photosystem I. This enables driving of the enzymatic reactions with electrons extracted from the photosynthetic electron transport chain. As a proof of concept, we have genetically fused the soluble catalytic domain of the cytochrome P450 CYP79A1, originating from the endoplasmic reticulum membranes of Sorghum bicolor, to a photosystem I subunit in the cyanobacterium Synechococcus sp. PCC 7002, thereby targeting it to the thylakoids. The engineered enzyme showed light-driven activity both in vivo and in vitro, demonstrating the possibility to achieve light-driven biosynthesis of high-value plant specialized metabolites in cyanobacteria.

The Tobacco mosaic virus Movement Protein Associates with but Does Not Integrate into Biological Membranes

Science.gov (United States)

Peiró, Ana; Martínez-Gil, Luis; Tamborero, Silvia; Pallás, Vicente

2014-01-01

ABSTRACT Plant positive-strand RNA viruses require association with plant cell endomembranes for viral translation and replication, as well as for intra- and intercellular movement of the viral progeny. The membrane association and RNA binding of the Tobacco mosaic virus (TMV) movement protein (MP) are vital for orchestrating the macromolecular network required for virus movement. A previously proposed topological model suggests that TMV MP is an integral membrane protein with two putative α-helical transmembrane (TM) segments. Here we tested this model using an experimental system that measured the efficiency with which natural polypeptide segments were inserted into the ER membrane under conditions approximating the in vivo situation, as well as in planta. Our results demonstrated that the two hydrophobic regions (HRs) of TMV MP do not span biological membranes. We further found that mutations to alter the hydrophobicity of the first HR modified membrane association and precluded virus movement. We propose a topological model in which the TMV MP HRs intimately associate with the cellular membranes, allowing maximum exposure of the hydrophilic domains of the MP to the cytoplasmic cellular components. IMPORTANCE To facilitate plant viral infection and spread, viruses encode one or more movement proteins (MPs) that interact with ER membranes. The present work investigated the membrane association of the 30K MP of Tobacco mosaic virus (TMV), and the results challenge the previous topological model, which predicted that the TMV MP behaves as an integral membrane protein. The current data provide greatly needed clarification of the topological model and provide substantial evidence that TMV MP is membrane associated only at the cytoplasmic face of the membrane and that neither of its domains is integrated into the membrane or translocated into the lumen. Understanding the topology of MPs in the ER is vital for understanding the role of the ER in plant virus transport

Integrating complex functions: coordination of nuclear pore complex assembly and membrane expansion of the nuclear envelope requires a family of integral membrane proteins.

Science.gov (United States)

Schneiter, Roger; Cole, Charles N

2010-01-01

The nuclear envelope harbors numerous large proteinaceous channels, the nuclear pore complexes (NPCs), through which macromolecular exchange between the cytosol and the nucleoplasm occurs. This double-membrane nuclear envelope is continuous with the endoplasmic reticulum and thus functionally connected to such diverse processes as vesicular transport, protein maturation and lipid synthesis. Recent results obtained from studies in Saccharomyces cerevisiae indicate that assembly of the nuclear pore complex is functionally dependent upon maintenance of lipid homeostasis of the ER membrane. Previous work from one of our laboratories has revealed that an integral membrane protein Apq12 is important for the assembly of functional nuclear pores. Cells lacking APQ12 are viable but cannot grow at low temperatures, have aberrant NPCs and a defect in mRNA export. Remarkably, these defects in NPC assembly can be overcome by supplementing cells with a membrane fluidizing agent, benzyl alcohol, suggesting that Apq12 impacts the flexibility of the nuclear membrane, possibly by adjusting its lipid composition when cells are shifted to a reduced temperature. Our new study now expands these findings and reveals that an essential membrane protein, Brr6, shares at least partially overlapping functions with Apq12 and is also required for assembly of functional NPCs. A third nuclear envelope membrane protein, Brl1, is related to Brr6, and is also required for NPC assembly. Because maintenance of membrane homeostasis is essential for cellular survival, the fact that these three proteins are conserved in fungi that undergo closed mitoses, but are not found in metazoans or plants, may indicate that their functions are performed by proteins unrelated at the primary sequence level to Brr6, Brl1 and Apq12 in cells that disassemble their nuclear envelopes during mitosis.

Membrane with integrated spacer

NARCIS (Netherlands)

Balster, J.H.; Stamatialis, Dimitrios; Wessling, Matthias

2010-01-01

Many membrane processes are severely influenced by concentration polarisation. Turbulence promoting spacers placed in between the membranes can reduce the diffusional resistance of concentration polarisation by inducing additional mixing. Electrodialysis (ED) used for desalination suffers from

The Primitive Thylakoid-Less Cyanobacterium Gloeobacter Is a Common Rock-Dwelling Organism.

Directory of Open Access Journals (Sweden)

Jan Mareš

Full Text Available Cyanobacteria are an ancient group of photosynthetic prokaryotes, which are significant in biogeochemical cycles. The most primitive among living cyanobacteria, Gloeobacter violaceus, shows a unique ancestral cell organization with a complete absence of inner membranes (thylakoids and an uncommon structure of the photosynthetic apparatus. Numerous phylogenetic papers proved its basal position among all of the organisms and organelles capable of plant-like photosynthesis (i.e., cyanobacteria, chloroplasts of algae and plants. Hence, G. violaceus has become one of the key species in evolutionary study of photosynthetic life. It also numbers among the most widely used organisms in experimental photosynthesis research. Except for a few related culture isolates, there has been little data on the actual biology of Gloeobacter, being relegated to an "evolutionary curiosity" with an enigmatic identity. Here we show that members of the genus Gloeobacter probably are common rock-dwelling cyanobacteria. On the basis of morphological, ultrastructural, pigment, and phylogenetic comparisons of available Gloeobacter strains, as well as on the basis of three new independent isolates and historical type specimen, we have produced strong evidence as to the close relationship of Gloeobacter to a long known rock-dwelling cyanobacterial morphospecies Aphanothece caldariorum. Our results bring new clues to solving the 40 year old puzzle of the true biological identity of Gloeobacter violaceus, a model organism with a high value in several biological disciplines. A probable broader distribution of Gloeobacter in common wet-rock habitats worldwide is suggested by our data, and its ecological meaning is discussed taking into consideration the background of cyanobacterial evolution. We provide observations of previously unknown genetic variability and phenotypic plasticity, which we expect to be utilized by experimental and evolutionary researchers worldwide.

Integration of oxygen membranes for oxygen production in cement plants

DEFF Research Database (Denmark)

Puig Arnavat, Maria; Søgaard, Martin; Hjuler, Klaus

2015-01-01

The present paper describes the integration of oxygen membranes in cement plants both from an energy, exergy and economic point of view. Different configurations for oxygen enrichment of the tertiary air for combustion in the pre-calciner and full oxy-fuel combustion in both pre-calciner and kiln...

Efficient ethanol recovery from fermentation broths with integrated distillation-membrane process

Science.gov (United States)

The energy demand of distillation-molecular sieve systems for ethanol recovery/dehydration can be significant, particularly for dilute solutions. An alternative process integrating vapor stripping (like a beer still) with vapor compression and a vapor permeation membrane separati...

Fabrication and characterization of an integrated ionic device from suspended polypyrrole and alamethicin-reconstituted lipid bilayer membranes

International Nuclear Information System (INIS)

Northcutt, Robert; Sundaresan, Vishnu-Baba

2012-01-01

Conducting polymers are electroactive materials that undergo conformal relaxation of the polymer backbone in the presence of an electrical field through ion exchange with solid or aqueous electrolytes. This conformal relaxation and the associated morphological changes make conducting polymers highly suitable for actuation and sensing applications. Among smart materials, bioderived active materials also use ion transport for sensing and actuation functions via selective ion transport. The transporter proteins extracted from biological cell membranes and reconstituted into a bilayer lipid membrane in bioderived active materials regulate ion transport for engineering functions. The protein transporter reconstituted in the bilayer lipid membrane is referred to as the bioderived membrane and serves as the active component in bioderived active materials. Inspired by the similarities in the physics of transduction in conducting polymers and bioderived active materials, an integrated ionic device is formed from the bioderived membrane and the conducting polymer membrane. This ionic device is fabricated into a laminated thin-film membrane and a common ion that can be processed by the bioderived and the conducting polymer membranes couple the ionic function of these two membranes. An integrated ionic device, fabricated from polypyrrole (PPy) doped with sodium dodecylbenzenesulfonate (NaDBS) and an alamethicin-reconstituted DPhPC bilayer lipid membrane, is presented in this paper. A voltage-gated sodium current regulates the electrochemical response in the PPy(DBS) layer. The integrated device is fabricated on silicon-based substrates through microfabrication, electropolymerization, and vesicle fusion, and ionic activity is characterized through electrochemical measurements. (paper)

Evaluation of the participation of ferredoxin in oxygen reduction in the photosynthetic electron transport chain of isolated pea thylakoids.

Science.gov (United States)

Kozuleva, Marina A; Ivanov, Boris N

2010-07-01

The contribution to reduction of oxygen by ferredoxin (Fd) to the overall reduction of oxygen in isolated pea thylakoids was studied in the presence of Fd versus Fd + NADP(+). The overall rate of electron transport was measured using a determination of Photosystem II quantum yield from chlorophyll fluorescence parameters, and the rate of oxidation of Fd was measured from the light-induced redox changes of Fd. At low light intensity, increasing Fd concentration from 5 to 30 microM in the absence of NADP(+) increased the proportion of oxygen reduction by Fd from 25-35 to 40-60% in different experiments. This proportion decreased with increasing light intensity. When NADP(+) was added in the presence of 15 microM Fd, which was optimal for the NADP(+) reduction rate, the participation of Fd in the reduction of oxygen was low, no more than 10%, and it also decreased with increasing light intensity. At high light intensity, the overall oxygen reduction rates in the presence of Fd + NADP(+) and in the presence of Fd alone were comparable. The significance of reduction of dioxygen either by water-soluble Fd or by the membrane-bound carriers of the photosynthetic electron transport chain for redox signaling under different light intensities is discussed.

CCR5 internalisation and signalling have different dependence on membrane lipid raft integrity.

Science.gov (United States)

Cardaba, Clara Moyano; Kerr, Jason S; Mueller, Anja

2008-09-01

The chemokine receptor, CCR5, acts as a co-receptor for human immunodeficiency virus entry into cells. CCR5 has been shown to be targeted to cholesterol- and sphingolipid-rich membrane microdomains termed lipid rafts or caveolae. Cholesterol is essential for CCL4 binding to CCR5 and for keeping the conformational integrity of the receptor. Filipin treatment leads to loss of caveolin-1 from the membrane and therefore to a collapse of the caveolae. We have found here that sequestration of membrane cholesterol with filipin did not affect receptor signalling, however a loss of ligand-induced internalisation of CCR5 was observed. Cholesterol extraction with methyl-beta-cyclodextrin (MCD) reduced signalling through CCR5 as measured by release of intracellular Ca(2+) and completely abolished the inhibition of forskolin-stimulated cAMP accumulation with no effect on internalisation. Pertussis toxin (PTX) treatment inhibited the intracellular release of calcium that is transduced via Galphai G-proteins. Depletion of cholesterol destroyed microdomains in the membrane and switched CCR5/G-protein coupling to a PTX-independent G-protein. We conclude that cholesterol in the membrane is essential for CCR5 signalling via the Galphai G-protein subunit, and that integrity of lipid rafts is not essential for effective CCR5 internalisation however it is crucial for proper CCR5 signal transduction via Galphai G-proteins.

Further Improvement and System Integration of High Temperature Polymer Electrolyte Membrane Fuel Cells

DEFF Research Database (Denmark)

Jensen, Jens Oluf; Li, Qingfeng

Polymer electrolyte membrane fuel cell (PEMFC) technology based on Nafion membranes can operate at temperatures around 80°C. The new development in the field is high temperature PEMFC for operation above 100°C, which has been successfully demonstrated through the previous EC Joule III and the 5th......, and system integration of the high temperature PEMFC. The strategic developments of the FURIM are in three steps: (1) further improvement of the high temperature polymer membranes and related materials; (2) development of technological units including fuel cell stack, hydrocarbon reformer, afterburner...... and power management system, that are compatible with the HT-PEMFC; and (3) integration of the HT-PEMFC stack with these compatible subunits. The main goal of the project is a 2kWel HT-PEMFC stack operating in a temperature range of 120-220°C, with a single cell performance target of 0.7 A/cm² at a cell...

Neumann and Neumann-Rosochatius integrable systems from membranes on AdS4 x S7

International Nuclear Information System (INIS)

Bozhilov, Plamen

2007-01-01

It is known that large class of classical string solutions in the type IIB AdS 5 x S 5 background is related to the Neumann and Neumann-Rosochatius integrable systems, including spiky strings and giant magnons. It is also interesting if these integrable systems can be associated with some membrane configurations in M-theory. We show here that this is indeed the case by presenting explicitly several types of membrane embedding in AdS 4 x S 7 with the searched properties

Integration of membrane filtration and photoelectrocatalysis using a TiO{sub 2}/carbon/Al{sub 2}O{sub 3} membrane for enhanced water treatment

Energy Technology Data Exchange (ETDEWEB)

Wang, Guanlong; Chen, Shuo, E-mail: shuochen@dlut.edu.cn; Yu, Hongtao; Quan, Xie

2015-12-15

Highlights: • Membrane filtration was integrated with photoelectrocatalysis for water treatment. • This integrated process (PECM) displays good antifouling capacity in NOMs removal. • PECM process enables efficient removal of chemical contaminants (e.g., RhB). • Enhanced charge separation of PECM process leads to its improved performance. - Abstract: Coupling membrane filtration with photocatalysis provides multifunction involving filtration and photocatalytic degradation for removing pollutants from water, but the performance of photocatalytic membrane is limited due to the quick recombination of photogenerated electron-holes in photocatalytic layer. Herein, a TiO{sub 2}/carbon/Al{sub 2}O{sub 3} membrane was designed and constructed through sequentially depositing graphitic carbon layer with good electro-conductivity and TiO{sub 2} nanoparticles layer with photocatalytic activity on Al{sub 2}O{sub 3} membrane support. When light irradiated on the membrane with a voltage supply, the photogenerated electrons could be drained from photocatalytic layer and separated with holes efficiently, thus endowing the membrane with photoelectrocatalytic function. Membrane performance tests indicated that the photoelectrocatalytic membrane filtration (PECM) showed improved removal of natural organic matters (NOMs) and permeate flux with increasing voltage supply. For PECM process at 1.0 V, its NOMs removal was 1.2 or 1.7 times higher than that of filtration with UV irradiation or filtration alone, and its stable permeate flux was 1.3 or 3 times higher than that of filtration with UV irradiation or filtration alone. Moreover, the PECM process exhibited special advantage in removing organic chemicals (e.g., Rhodamine B), which displayed 1.3 or 3 times higher removal than that of filtration with UV irradiation or filtration alone.

A Printed Equilibrium Dialysis Device with Integrated Membranes for Improved Binding Affinity Measurements.

Science.gov (United States)

Pinger, Cody W; Heller, Andrew A; Spence, Dana M

2017-07-18

Equilibrium dialysis is a simple and effective technique used for investigating the binding of small molecules and ions to proteins. A three-dimensional (3D) printer was used to create a device capable of measuring binding constants between a protein and a small ion based on equilibrium dialysis. Specifically, the technology described here enables the user to customize an equilibrium dialysis device to fit their own experiments by choosing membranes of various material and molecular-weight cutoff values. The device has dimensions similar to that of a standard 96-well plate, thus being amenable to automated sample handlers and multichannel pipettes. The device consists of a printed base that hosts multiple windows containing a porous regenerated-cellulose membrane with a molecular-weight cutoff of ∼3500 Da. A key step in the fabrication process is a print-pause-print approach for integrating membranes directly into the windows subsequently inserted into the base. The integrated membranes display no leaking upon placement into the base. After characterizing the system's requirements for reaching equilibrium, the device was used to successfully measure an equilibrium dissociation constant for Zn 2+ and human serum albumin (K d = (5.62 ± 0.93) × 10 -7 M) under physiological conditions that is statistically equal to the constants reported in the literature.

Membrane potential and microsecond to millisecond delayed light emission after a single excitation flash in isolated chloroplasts

International Nuclear Information System (INIS)

Jursinic, P.; Govindjee; Wraight, C.A.

1978-01-01

The effect of light-induced and salt-jump induced membrane potential on microsecond and millisecond delayed light emission from chloroplasts, following a single 10 ns flash, have been studied. Microsecond delayed light emission is shown to be independent of the membrane potential contrary to proposals that the activation energy for delayed light emission can be modulated by transmembrane electric fields. This result is discussed in terms of the possible origin of this short-lived emission. Millisecond delayed light after a single excitation flash is enhanced by membrane potential only if a proton gradient is present. By measuring changes in ms delayed light caused by simultaneous injection of KCl and Na-benzoate (which creates a proton gradient) in the presence of valinomycin, the light-induced potential generated across the thylakoid membrane by a single excitation flash was calibrated and found to be 128 +- 10 mV in agreement with the recent measurements of Zickler and Witt, (FEBS Lett. 66, 142-148 (1976)), based on voltage-dependent ionophores. It is concluded that the secondary charges that give rise to ms delayed light, after a single flash, do not fully span the membrane. (author)

The cell-free integration of a polytopic mitochondrial membrane protein into liposomes occurs cotranslationally and in a lipid-dependent manner.

Directory of Open Access Journals (Sweden)

Ashley R Long

Full Text Available The ADP/ATP Carrier (AAC is the most abundant transporter of the mitochondrial inner membrane. The central role that this transporter plays in cellular energy production highlights the importance of understanding its structure, function, and the basis of its pathologies. As a means of preparing proteoliposomes for the study of membrane proteins, several groups have explored the use of cell-free translation systems to facilitate membrane protein integration directly into preformed unilamellar vesicles without the use of surfactants. Using AAC as a model, we report for the first time the detergent-free reconstitution of a mitochondrial inner membrane protein into liposomes using a wheat germ-based in vitro translation system. Using a host of independent approaches, we demonstrate the efficient integration of AAC into vesicles with an inner membrane-mimetic lipid composition and, more importantly, that the integrated AAC is functionally active in transport. By adding liposomes at different stages of the translation reaction, we show that this direct integration is obligatorily cotranslational, and by synthesizing stable ribosome-bound nascent chain intermediates, we show that the nascent AAC polypeptide interacts with lipid vesicles while ribosome-bound. Finally, we show that the presence of the phospholipid cardiolipin in the liposomes specifically enhances AAC translation rate as well as the efficiency of vesicle association and integration. In light of these results, the possible mechanisms of liposome-assisted membrane protein integration during cell-free translation are discussed with respect to the mode of integration and the role of specific lipids.

An integrated membrane system for the biocatalytic production of 3′-sialyllactose from dairy by-products

DEFF Research Database (Denmark)

Luo, Jianquan; Nordvang, Rune Thorbjørn; Morthensen, Sofie Thage

2014-01-01

An integrated membrane system was investigated for the production of 30-sialyllactose by an engineered sialidase using casein glycomacropeptide (CGMP) and lactose as substrates. CGMP was purified by ultrafiltration (UF) to remove any small molecules present and then an enzymatic membrane reactor ...

Diffusion of Integral Membrane Proteins in Protein-Rich Membranes

DEFF Research Database (Denmark)

Javanainen, Matti; Martinez-Seara, Hector; Metzler, Ralf

2017-01-01

of being protein-poor, native cell membranes are extremely crowded with proteins. On the basis of extensive molecular simulations, we here demonstrate that protein crowding of the membrane at physiological levels leads to deviations from the SD relation and to the emergence of a stronger Stokes......-like dependence D ∝ 1/R. We propose that this 1/R law mainly arises due to geometrical factors: smaller proteins are able to avoid confinement effects much better than their larger counterparts. The results highlight that the lateral dynamics in the crowded setting found in native membranes is radically different......The lateral diffusion of embedded proteins along lipid membranes in protein-poor conditions has been successfully described in terms of the Saffman-Delbrück (SD) model, which predicts that the protein diffusion coefficient D is weakly dependent on its radius R as D ∝ ln(1/R). However, instead...

Integrated antibacterial and antifouling surfaces via cross-linking chitosan-g-eugenol/zwitterionic copolymer on electrospun membranes.

Science.gov (United States)

Li, Zhenguang; Hu, Wenhong; Zhao, Yunhui; Ren, Lixia; Yuan, Xiaoyan

2018-04-27

Integrated antibacterial and antifouling surfaces in favor of avoiding implant-related infections are necessarily required for biomaterials when they contact with the body fluid. In this work, an antibacterial and antifouling membrane was developed via cross-linking chitosan-g-eugenol and the zwitterionic copolymer poly(sulfobetaine methylacrylate-co-2-aminoethyl methacrylate) on the electrospun polycarbonate urethane substrate using genipin as a cross-linker. Antibacterial assays demonstrated that the prepared membranes had efficient antibacterial activity with 92.8 ± 2.5% and 95.2 ± 1.3% growth inhibition rates against Escherichia coli and Staphylococcus aureus, respectively. The investigations on antifouling activity and hemocompatibility of the membranes showed significant resistances to bacterial attachment, non-specific protein adsorption and platelet adhesion, and presented lower hemolytic activity and good anticoagulant activity as well. Moreover, cell culture assays indicated that the prepared membranes exerted no obvious cytotoxicity with more than 80% of relative L929 fibroblast viability. Therefore, the membranes with integrated antibacterial and antifouling properties could be potentially applied in promising indwelling devices. Copyright © 2018 Elsevier B.V. All rights reserved.

FAD oxidizes the ERO1-PDI electron transfer chain: The role of membrane integrity

International Nuclear Information System (INIS)

Papp, Eszter; Nardai, Gabor; Mandl, Jozsef; Banhegyi, Gabor; Csermely, Peter

2005-01-01

The molecular steps of the electron transfer in the endoplasmic reticulum from the secreted proteins during their oxidation are relatively unknown. We present here that flavine adenine dinucleotide (FAD) is a powerful oxidizer of the oxidoreductase system, Ero1 and PDI, besides the proteins of rat liver microsomes and HepG2 hepatoma cells. Inhibition of FAD transport hindered the action of FAD. Microsomal membrane integrity was mandatory for all FAD-related oxidation steps downstream of Ero1. The PDI inhibitor bacitracin could inhibit FAD-mediated oxidation of microsomal proteins and PDI, but did not hinder the FAD-driven oxidation of Ero1. Our data demonstrated that Ero1 can utilize FAD as an electron acceptor and that FAD-driven protein oxidation goes through the Ero1-PDI pathway and requires the integrity of the endoplasmic reticulum membrane. Our findings prompt further studies to elucidate the membrane-dependent steps of PDI oxidation and the role of FAD in redox folding

Structure determination of an integral membrane protein at room temperature from crystals in situ

International Nuclear Information System (INIS)

Axford, Danny; Foadi, James; Hu, Nien-Jen; Choudhury, Hassanul Ghani; Iwata, So; Beis, Konstantinos; Evans, Gwyndaf; Alguel, Yilmaz

2015-01-01

The X-ray structure determination of an integral membrane protein using synchrotron diffraction data measured in situ at room temperature is demonstrated. The structure determination of an integral membrane protein using synchrotron X-ray diffraction data collected at room temperature directly in vapour-diffusion crystallization plates (in situ) is demonstrated. Exposing the crystals in situ eliminates manual sample handling and, since it is performed at room temperature, removes the complication of cryoprotection and potential structural anomalies induced by sample cryocooling. Essential to the method is the ability to limit radiation damage by recording a small amount of data per sample from many samples and subsequently assembling the resulting data sets using specialized software. The validity of this procedure is established by the structure determination of Haemophilus influenza TehA at 2.3 Å resolution. The method presented offers an effective protocol for the fast and efficient determination of membrane-protein structures at room temperature using third-generation synchrotron beamlines

Structure determination of an integral membrane protein at room temperature from crystals in situ

Energy Technology Data Exchange (ETDEWEB)

Axford, Danny [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Foadi, James [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Imperial College London, London SW7 2AZ (United Kingdom); Hu, Nien-Jen; Choudhury, Hassanul Ghani [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Imperial College London, London SW7 2AZ (United Kingdom); Rutherford Appleton Laboratory, Oxfordshire OX11 0FA (United Kingdom); Iwata, So [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Imperial College London, London SW7 2AZ (United Kingdom); Rutherford Appleton Laboratory, Oxfordshire OX11 0FA (United Kingdom); Kyoto University, Kyoto 606-8501 (Japan); Beis, Konstantinos [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Imperial College London, London SW7 2AZ (United Kingdom); Rutherford Appleton Laboratory, Oxfordshire OX11 0FA (United Kingdom); Evans, Gwyndaf, E-mail: gwyndaf.evans@diamond.ac.uk [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Alguel, Yilmaz, E-mail: gwyndaf.evans@diamond.ac.uk [Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire OX11 0DE (United Kingdom); Imperial College London, London SW7 2AZ (United Kingdom); Rutherford Appleton Laboratory, Oxfordshire OX11 0FA (United Kingdom)

2015-05-14

The X-ray structure determination of an integral membrane protein using synchrotron diffraction data measured in situ at room temperature is demonstrated. The structure determination of an integral membrane protein using synchrotron X-ray diffraction data collected at room temperature directly in vapour-diffusion crystallization plates (in situ) is demonstrated. Exposing the crystals in situ eliminates manual sample handling and, since it is performed at room temperature, removes the complication of cryoprotection and potential structural anomalies induced by sample cryocooling. Essential to the method is the ability to limit radiation damage by recording a small amount of data per sample from many samples and subsequently assembling the resulting data sets using specialized software. The validity of this procedure is established by the structure determination of Haemophilus influenza TehA at 2.3 Å resolution. The method presented offers an effective protocol for the fast and efficient determination of membrane-protein structures at room temperature using third-generation synchrotron beamlines.

Towards a fully integrated indium-phosphide membrane on silicon photonics platform

NARCIS (Netherlands)

van Engelen, J.P.; Pogoretskiy, V.; Smit, M.K.; van der Tol, J.J.G.M.; Jiao, Y.

2017-01-01

Recently a uni-traveling-carrier photodetector with high speed (> 67GHz) and a high-gain optical amplifier (110/cm at 4 kA/cm2) have been demonstrated using the InP membrane-on-Silicon (IMOS) integration technology. Passives in IMOS have shown features comparable to SOI platforms due to the tight

Integrated forward osmosis-membrane distillation process for human urine treatment.

Science.gov (United States)

Liu, Qianliang; Liu, Caihong; Zhao, Lei; Ma, Weichao; Liu, Huiling; Ma, Jun

2016-03-15

This study demonstrated a forward osmosis-membrane distillation (FO-MD) hybrid system for real human urine treatment. A series of NaCl solutions at different concentrations were adopted for draw solutions in FO process, which were also the feed solutions of MD process. To establish a stable and continuous integrated FO-MD system, individual FO process with different NaCl concentrations and individual direct contact membrane distillation (DCMD) process with different feed temperatures were firstly investigated separately. Four stable equilibrium conditions were obtained from matching the water transfer rates of individual FO and MD processes. It was found that the integrated system is stable and sustainable when the water transfer rate of FO subsystem is equal to that of MD subsystem. The rejections to main contaminants in human urine were also investigated. Although individual FO process had relatively high rejection to Total Organic Carbon (TOC), Total Nitrogen (TN) and Ammonium Nitrogen (NH4(+)-N) in human urine, these contaminants could also accumulate in draw solution after long term performance. The MD process provided an effective rejection to contaminants in draw solution after FO process and the integrated system revealed nearly complete rejection to TOC, TN and NH4(+)-N. This work provided a potential treatment process for human urine in some fields such as water regeneration in space station and water or nutrient recovery from source-separated urine. Copyright © 2016 Elsevier Ltd. All rights reserved.

A continuous membrane microbioreactor system for development of integrated pectin modification and separation processes

DEFF Research Database (Denmark)

Zainal Alam, Muhd Nazrul Hisham Bin; Pinelo, Manuel; Samanta, Kama

2011-01-01

present a continuous membrane microbioreactor prototype for development of enzyme catalyzed degradation of pectin. Membrane reactors are becoming increasingly important for the novel ‘biorefining’ type of processes that either require product removal to avoid product inhibition or rest on partial...... hydrolysis of the substrate to obtain e.g. value-added oligosaccharides from complex biopolymers. The microbioreactor prototype was fabricated from poly(methylmethacrylate) (PMMA) and poly(dimethylsiloxane) (PDMS) and designed as a loop reactor (working volume approximately 190μL) integrated...... with a regenerated cellulose membrane for separation of low molecular weight products. The main technical considerations and challenges related to establishing the continuous membrane microbioreactor are discussed. The workability of the prototype was validated by comparing the process data at microscale to those...

Characterization of chloroplast phosphoproteins controlling manganese use efficiency using quantitative proteomics

DEFF Research Database (Denmark)

Petersen, Jørgen; Sprenger, Richard Remko; Rogowska-Wrzesinska, Adelina

Manganese is important for molecular functions in plants, i.e. as a co-factor in enzymes and in the oxygen evolving complex of photosystem II, located like most of the photosynthetic machinery, in the thylakoid membranes of chloroplasts. Soils that lack plant available micronutrients such as mang......Manganese is important for molecular functions in plants, i.e. as a co-factor in enzymes and in the oxygen evolving complex of photosystem II, located like most of the photosynthetic machinery, in the thylakoid membranes of chloroplasts. Soils that lack plant available micronutrients...... involved in manganese use efficiency, focusing on the phosphoproteome from thylakoid preparations from two barley genotypes, manganese efficient (Vanessa) and inefficient (Antonia) genotype. Experimental: By monitoring the photosynthetic efficiency (Fv/Fm) a decline in activity is observed as a consequence...

Risk assessment of Giardia from a full scale MBR sewage treatment plant caused by membrane integrity failure.

Science.gov (United States)

Zhang, Yu; Chen, Zhimin; An, Wei; Xiao, Shumin; Yuan, Hongying; Zhang, Dongqing; Yang, Min

2015-04-01

Membrane bioreactors (MBR) are highly efficient at intercepting particles and microbes and have become an important technology for wastewater reclamation. However, many pathogens can accumulate in activated sludge due to the long residence time usually adopted in MBR, and thus may pose health risks when membrane integrity problems occur. This study presents data from a survey on the occurrence of water-borne Giardia pathogens in reclaimed water from a full-scale wastewater treatment plant with MBR experiencing membrane integrity failure, and assessed the associated risk for green space irrigation. Due to membrane integrity failure, the MBR effluent turbidity varied between 0.23 and 1.90 NTU over a period of eight months. Though this turbidity level still met reclaimed water quality standards (≤5 NTU), Giardia were detected at concentrations of 0.3 to 95 cysts/10 L, with a close correlation between effluent turbidity and Giardia concentration. All β-giardin gene sequences of Giardia in the WWTP influents were genotyped as Assemblages A and B, both of which are known to infect humans. An exponential dose-response model was applied to assess the risk of infection by Giardia. The risk in the MBR effluent with chlorination was 9.83×10(-3), higher than the acceptable annual risk of 1.0×10(-4). This study suggested that membrane integrity is very important for keeping a low pathogen level, and multiple barriers are needed to ensure the biological safety of MBR effluent. Copyright © 2015. Published by Elsevier B.V.

Subcellular localization and logistics of integral membrane protein biogenesis in Escherichia coli.

Science.gov (United States)

Bogdanov, Mikhail; Aboulwafa, Mohammad; Saier, Milton H

2013-01-01

Transporters catalyze entry and exit of molecules into and out of cells and organelles, and protein-lipid interactions influence their activities. The bacterial phosphoenolpyruvate: sugar phosphotransferase system (PTS) catalyzes transport-coupled sugar phosphorylation as well as nonvectorial sugar phosphorylation in the cytoplasm. The vectorial process is much more sensitive to the lipid environment than the nonvectorial process. Moreover, cytoplasmic micellar forms of these enzyme-porters have been identified, and non-PTS permeases have similarly been shown to exist in 'soluble' forms. The latter porters exhibit lipid-dependent activities and can adopt altered topologies by simply changing the lipid composition. Finally, intracellular membranes and vesicles exist in Escherichia coli leading to the following unanswered questions: (1) what determines whether a PTS permease catalyzes vectorial or nonvectorial sugar phosphorylation? (2) How do phospholipids influence relative amounts of the plasma membrane, intracellular membrane, inner membrane-derived vesicles and cytoplasmic micelles? (3) What regulates the route(s) of permease insertion and transfer into and between the different subcellular sites? (4) Do these various membranous forms have distinct physiological functions? (5) What methods should be utilized to study the biogenesis and interconversion of these membranous structures? While research concerning these questions is still in its infancy, answers will greatly enhance our understanding of protein-lipid interactions and how they control the activities, conformations, cellular locations and biogenesis of integral membrane proteins. Copyright © 2013 S. Karger AG, Basel.

MapA, an iron-regulated, cytoplasmic membrane protein in the cyanobacterium Synechococcus sp. strain PCC7942.

Science.gov (United States)

Webb, R; Troyan, T; Sherman, D; Sherman, L A

1994-08-01

Growth of Synechococcus sp. strain PCC 7942 in iron-deficient media leads to the accumulation of an approximately 34-kDa protein. The gene encoding this protein, mapA (membrane-associated protein A), has been cloned and sequenced (GenBank accession number, L01621). The mapA transcript is not detectable in normally grown cultures but is stably accumulated by cells grown in iron-deficient media. However, the promoter sequence for this gene does not resemble other bacterial iron-regulated promoters described to date. The carboxyl-terminal region of the derived amino acid sequence of MapA resembles bacterial proteins involved in iron acquisition, whereas the amino-terminal end of MapA has a high degree of amino acid identity with the abundant, chloroplast envelope protein E37. An approach employing improved cellular fractionation techniques as well as electron microscopy and immunocytochemistry was essential in localizing MapA protein to the cytoplasmic membrane of Synechococcus sp. strain PCC 7942. When these cells were grown under iron-deficient conditions, a significant fraction of MapA could also be localized to the thylakoid membranes.

Monitoring changes in membrane polarity, membrane integrity, and intracellular ion concentrations in Streptococcus pneumoniae using fluorescent dyes.

Science.gov (United States)

Clementi, Emily A; Marks, Laura R; Roche-Håkansson, Hazeline; Håkansson, Anders P

2014-02-17

Membrane depolarization and ion fluxes are events that have been studied extensively in biological systems due to their ability to profoundly impact cellular functions, including energetics and signal transductions. While both fluorescent and electrophysiological methods, including electrode usage and patch-clamping, have been well developed for measuring these events in eukaryotic cells, methodology for measuring similar events in microorganisms have proven more challenging to develop given their small size in combination with the more complex outer surface of bacteria shielding the membrane. During our studies of death-initiation in Streptococcus pneumoniae (pneumococcus), we wanted to elucidate the role of membrane events, including changes in polarity, integrity, and intracellular ion concentrations. Searching the literature, we found that very few studies exist. Other investigators had monitored radioisotope uptake or equilibrium to measure ion fluxes and membrane potential and a limited number of studies, mostly in Gram-negative organisms, had seen some success using carbocyanine or oxonol fluorescent dyes to measure membrane potential, or loading bacteria with cell-permeant acetoxymethyl (AM) ester versions of ion-sensitive fluorescent indicator dyes. We therefore established and optimized protocols for measuring membrane potential, rupture, and ion-transport in the Gram-positive organism S. pneumoniae. We developed protocols using the bis-oxonol dye DiBAC4(3) and the cell-impermeant dye propidium iodide to measure membrane depolarization and rupture, respectively, as well as methods to optimally load the pneumococci with the AM esters of the ratiometric dyes Fura-2, PBFI, and BCECF to detect changes in intracellular concentrations of Ca(2+), K(+), and H(+), respectively, using a fluorescence-detection plate reader. These protocols are the first of their kind for the pneumococcus and the majority of these dyes have not been used in any other bacterial

Further Improvement and System Integration of High Temperature Polymer Electrolyte Membrane Fuel Cells

DEFF Research Database (Denmark)

Li, Qingfeng; Jensen, Jens Oluf

The new development in the field of polymer electrolyte membrane fuel cell (PEMFC) is high temperature PEMFC for operation above 100°C, which has been successfully demonstrated through the previous EC Joule III and the 5th framework programme. New challenges are encountered, bottlenecks for the new...... technology have been identified, and new concepts and solutions have been provisionally identified. FURIM is directed at tackling these key issues by concentrating on the further materials development, compatible technologies, and system integration of the high temperature PEMFC. The strategic developments...... of the FURIM are in three steps: (1) further improvement of the high temperature polymer membranes and related materials; (2) development of technological units including fuel cell stack, hydrocarbon reformer and afterburner, that are compatible with the HT-PEMFC; and (3) integration of the HT-PEMFC stack...

Seawater desalination with solar-energy-integrated vacuum membrane distillation system

Directory of Open Access Journals (Sweden)

Fang Wang

2017-03-01

Full Text Available This study designed and tested a novel type of solar-energy-integrated vacuum membrane distillation (VMD system for seawater desalination under actual environmental conditions in Wuhan, China. The system consists of eight parts: a seawater tank, solar collector, solar cooker, inclined VMD evaporator, circulating water vacuum pump, heat exchanger, fresh water tank, and brine tank. Natural seawater was used as feed and a hydrophobic hollow-fiber membrane module was used to improve seawater desalination. The experiment was conducted during a typical summer day. Results showed that when the highest ambient temperature was 33 °C, the maximum value of the average solar intensity was 1,080 W/m2. The system was able to generate 36 kg (per m2 membrane module distilled fresh water during 1 day (7:00 am until 6:00 pm, the retention rate was between 99.67 and 99.987%, and electrical conductivity was between 0.00276 and 0.0673 mS/cm. The average salt rejection was over 90%. The proposed VMD system shows favorable potential application in desalination of brackish waters or high-salt wastewater treatment, as well.

Integrated Wireless Monitoring and Control System in Reverse Osmosis Membrane Desalination Plants

Directory of Open Access Journals (Sweden)

Al Haji Ahmad

2015-01-01

Full Text Available The operational processes of the Reverse Osmosis (RO membrane desalination plants require continuous monitoring through the constant attendance of operators to ensure proper productivity and minimize downtime and prevent membrane failure. Therefore, the plant must be equipped with a control system that monitors and controls the operational variables. Monitoring and controlling the affecting parameters are critical to the evaluation of the performance of the desalination plant, which will help the operator find and resolve problems immediately. Therefore, this paper was aimed at developing an RO unit by utilizing a wireless sensor network (WSN system. Hence, an RO pilot plant with a feed capacity of 1.2 m3/h was utilized, commissioned, and tested in Kuwait to assess and verify the performance of the integrated WSN in RO membrane desalination system. The investigated system allowed the operators to remotely monitor the operational process of the RO system. The operational data were smoothly recorded and monitored. Furthermore, the technical problems were immediately determined, which reduced the time and effort in rectifying the technical problems relevant to the RO performance. The manpower requirements of such treatment system were dramatically reduced by about 50%. Based on a comparison between manual and wireless monitoring operational processes, the availability of the integrated RO unit with a wireless monitoring was increased by 10%

Nanowire-integrated microporous silicon membrane for continuous fluid transport in micro cooling device

International Nuclear Information System (INIS)

So, Hongyun; Pisano, Albert P.; Cheng, Jim C.

2013-01-01

We report an efficient passive micro pump system combining the physical properties of nanowires and micropores. This nanowire-integrated microporous silicon membrane was created to feed coolant continuously onto the surface of the wick in a micro cooling device to ensure it remains hydrated and in case of dryout, allow for regeneration of the system. The membrane was fabricated by photoelectrochemical etching to form micropores followed by hydrothermal growth of nanowires. This study shows a promising approach to address thermal management challenges for next generation electronic devices with absence of external power

Recovery of flavonoids from orange press liquor by an integrated membrane process.

Science.gov (United States)

Cassano, Alfredo; Conidi, Carmela; Ruby-Figueroa, René

2014-08-11

Orange press liquor is a by-product generated by the citrus processing industry containing huge amounts of natural phenolic compounds with recognized antioxidant activity. In this work, an integrated membrane process for the recovery of flavonoids from orange press liquors was investigated on a laboratory scale. The liquor was previously clarified by ultrafiltration (UF) in selected operating conditions by using hollow fiber polysulfone membranes. Then, the clarified liquor with a total soluble solids (TSS) content of 10 g·100 g-1 was pre-concentrated by nanofiltration (NF) up to 32 g TSS 100 g-1 by using a polyethersulfone spiral-wound membrane. A final concentration step, up to 47 g TSS 100 g-1, was performed by using an osmotic distillation (OD) apparatus equipped with polypropylene hollow fiber membranes. Suspended solids were completely removed in the UF step producing a clarified liquor containing most part of the flavonoids of the original press liquor due to the low rejection of the UF membrane towards these compounds. Flavanones and anthocyanins were highly rejected by the NF membrane, producing a permeate stream with a TSS content of 4.5 g·100 g-1. An increasing of both the flavanones and anthocyanins concentration was observed in the NF retentate by increasing the volume reduction factor (VRF). The final concentration of flavonoids by OD produced a concentrated solution of interest for nutraceutical and pharmaceutical applications.

Recovery of Flavonoids from Orange Press Liquor by an Integrated Membrane Process

Directory of Open Access Journals (Sweden)

Alfredo Cassano

2014-08-01

Full Text Available Orange press liquor is a by-product generated by the citrus processing industry containing huge amounts of natural phenolic compounds with recognized antioxidant activity. In this work, an integrated membrane process for the recovery of flavonoids from orange press liquors was investigated on a laboratory scale. The liquor was previously clarified by ultrafiltration (UF in selected operating conditions by using hollow fiber polysulfone membranes. Then, the clarified liquor with a total soluble solids (TSS content of 10 g·100 g−1 was pre-concentrated by nanofiltration (NF up to 32 g TSS 100 g−1 by using a polyethersulfone spiral-wound membrane. A final concentration step, up to 47 g TSS 100 g−1, was performed by using an osmotic distillation (OD apparatus equipped with polypropylene hollow fiber membranes. Suspended solids were completely removed in the UF step producing a clarified liquor containing most part of the flavonoids of the original press liquor due to the low rejection of the UF membrane towards these compounds. Flavanones and anthocyanins were highly rejected by the NF membrane, producing a permeate stream with a TSS content of 4.5 g·100 g−1. An increasing of both the flavanones and anthocyanins concentration was observed in the NF retentate by increasing the volume reduction factor (VRF. The final concentration of flavonoids by OD produced a concentrated solution of interest for nutraceutical and pharmaceutical applications.

Proteomic Analysis of Chloroplast-to-Chromoplast Transition in Tomato Reveals Metabolic Shifts Coupled with Disrupted Thylakoid Biogenesis Machinery and Elevated Energy-Production Components1[W

Science.gov (United States)

Barsan, Cristina; Zouine, Mohamed; Maza, Elie; Bian, Wanping; Egea, Isabel; Rossignol, Michel; Bouyssie, David; Pichereaux, Carole; Purgatto, Eduardo; Bouzayen, Mondher; Latché, Alain; Pech, Jean-Claude

2012-01-01

A comparative proteomic approach was performed to identify differentially expressed proteins in plastids at three stages of tomato (Solanum lycopersicum) fruit ripening (mature-green, breaker, red). Stringent curation and processing of the data from three independent replicates identified 1,932 proteins among which 1,529 were quantified by spectral counting. The quantification procedures have been subsequently validated by immunoblot analysis of six proteins representative of distinct metabolic or regulatory pathways. Among the main features of the chloroplast-to-chromoplast transition revealed by the study, chromoplastogenesis appears to be associated with major metabolic shifts: (1) strong decrease in abundance of proteins of light reactions (photosynthesis, Calvin cycle, photorespiration) and carbohydrate metabolism (starch synthesis/degradation), mostly between breaker and red stages and (2) increase in terpenoid biosynthesis (including carotenoids) and stress-response proteins (ascorbate-glutathione cycle, abiotic stress, redox, heat shock). These metabolic shifts are preceded by the accumulation of plastid-encoded acetyl Coenzyme A carboxylase D proteins accounting for the generation of a storage matrix that will accumulate carotenoids. Of particular note is the high abundance of proteins involved in providing energy and in metabolites import. Structural differentiation of the chromoplast is characterized by a sharp and continuous decrease of thylakoid proteins whereas envelope and stroma proteins remain remarkably stable. This is coincident with the disruption of the machinery for thylakoids and photosystem biogenesis (vesicular trafficking, provision of material for thylakoid biosynthesis, photosystems assembly) and the loss of the plastid division machinery. Altogether, the data provide new insights on the chromoplast differentiation process while enriching our knowledge of the plant plastid proteome. PMID:22908117

Role for ribosome-associated complex and stress-seventy subfamily B (RAC-Ssb) in integral membrane protein translation.

Science.gov (United States)

Acosta-Sampson, Ligia; Döring, Kristina; Lin, Yuping; Yu, Vivian Y; Bukau, Bernd; Kramer, Günter; Cate, Jamie H D

2017-12-01

Targeting of most integral membrane proteins to the endoplasmic reticulum is controlled by the signal recognition particle, which recognizes a hydrophobic signal sequence near the protein N terminus. Proper folding of these proteins is monitored by the unfolded protein response and involves protein degradation pathways to ensure quality control. Here, we identify a new pathway for quality control of major facilitator superfamily transporters that occurs before the first transmembrane helix, the signal sequence recognized by the signal recognition particle, is made by the ribosome. Increased rates of translation elongation of the N-terminal sequence of these integral membrane proteins can divert the nascent protein chains to the ribosome-associated complex and stress-seventy subfamily B chaperones. We also show that quality control of integral membrane proteins by ribosome-associated complex-stress-seventy subfamily B couples translation rate to the unfolded protein response, which has implications for understanding mechanisms underlying human disease and protein production in biotechnology. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Action spectrum for inhibition by ultraviolet radiation of photosystem 2 activity in spinach thylakoids

International Nuclear Information System (INIS)

Bornman, J.F.; Bjoern, L.O.; Aakerlund, H.-E.

1984-01-01

The effect of ultraviolet (UV) radiation (half-band width 10 nm) in the wavelength range 248-340 nm on chlorophyll fluorescence from a thin layer of spinach thylakoid suspension was investigated. It was found that the parameter most sensitive to UV radiation was the rise time of variable fluorescence. The increase in rise time was proportional to UV photon fluence and was used for the determination of an action spectrum. The action spectrum falls off from a maximum at ca. 275 nm towards longer wavelengths and rises from a minimum at 260 nm towards shorter wavelengths. The results also suggest that the UV inhibition is mainly on the PS 2 oxidizing side. Possibly damage is also inflicted to the PS 2 reaction center. (orig.)

Chloroplastic responses of ponderosa pine (Pinus ponderosa) seedlings to ozone exposure.

Science.gov (United States)

Anderson, Paul D; Palmer, Brent; Houpis, James L J; Smith, Mary K; Pushnik, James C

2003-06-01

Integrity of chloroplast membranes is essential to photosynthesis. Loss of thylakoid membrane integrity has been proposed as a consequence of ozone (O(3)) exposure and therefore may be a mechanistic basis for decreased photosynthetic rates commonly associated with ozone exposure. To investigate this hypothesis, Pinus ponderosa seedlings were exposed to ambient air or ozone concentrations maintained at 0.15 or 0.30 microliter l(-1) for 10 h day(-1) for 51 days during their second growing season. Over the course of the study, foliage samples were periodically collected for thylakoid membrane, chlorophyll and protein analyses. Additionally, gas-exchange measurements were made in conjunction with foliage sampling to verify that observed chloroplastic responses were associated with ozone-induced changes in photosynthesis. Needles exposed to elevated ozone exhibited decreases in chlorophyll a and b content. The decreases were dependent on the duration and intensity of ozone exposure. When based on equal amounts of chlorophyll, ozone-exposed sample tissue exhibited an increase in total protein. When based on equal amounts of protein, ozone-exposed samples exhibited an increase in 37 kDa proteins, possibly consisting of breakdown products, and a possible decrease in 68 kDa proteins, Rubisco small subunit. There was also a change in the ratio of Photosystem I protein complexes CPI and CPII that may have contributed to decreased photosynthesis. Net photosynthetic rates were decreased in the high ozone treatment suggesting that observed structural and biochemical changes in the chloroplast were associated with alterations of the photosynthetic process.

Integrated gasification gas combined cycle plant with membrane reactors: Technological and economical analysis

International Nuclear Information System (INIS)

Amelio, Mario; Morrone, Pietropaolo; Gallucci, Fausto; Basile, Angelo

2007-01-01

In the present work, the capture and storage of carbon dioxide from the fossil fuel power plant have been considered. The main objective was to analyze the thermodynamic performances and the technological aspects of two integrated gasification gas combined cycle plants (IGCC), as well as to give a forecast of the investment costs for the plants and the resulting energy consumptions. The first plant considered is an IGCC* plant (integrated gasification gas combined cycle plant with traditional shift reactors) characterized by the traditional water gas shift reactors and a CO 2 physical adsorption system followed by the power section. The second one is an IGCC M plant (integrated gasification gas combined cycle plant with membrane reactor) where the coal thermal input is the same as the first one, but the traditional shift reactors and the physical adsorption unit are replaced by catalytic palladium membrane reactors (CMR). In the present work, a mono-dimensional computational model of the membrane reactor was proposed to simulate and evaluate the capability of the IGCC M plant to capture carbon dioxide. The energetic performances, efficiency and net power of the IGCC* and IGCC M plants were, thus, compared, assuming as standard a traditional IGCC plant without carbon dioxide capture. The economical aspects of the three plants were compared through an economical analysis. Since the IGCC* and IGCC M plants have additional costs related to the capture and disposal of the carbon dioxide, a Carbon Tax (adopted in some countries like Sweden) proportional to the number of kilograms of carbon dioxide released in the environment was assumed. According to the economical analysis, the IGCC M plant proved to be more convenient than the IGCC* one

Photosystem II repair and plant immunity: Lessons learned from Arabidopsis mutant lacking the THYLAKOID LUMEN PROTEIN 18.3

Directory of Open Access Journals (Sweden)

Sari eJärvi

2016-03-01

Full Text Available Chloroplasts play an important role in the cellular sensing of abiotic and biotic stress. Signals originating from photosynthetic light reactions, in the form of redox and pH changes, accumulation of reactive oxygen and electrophile species or stromal metabolites are of key importance in chloroplast retrograde signaling. These signals initiate plant acclimation responses to both abiotic and biotic stresses. To reveal the molecular responses activated by rapid fluctuations in growth light intensity, gene expression analysis was performed with Arabidopsis thaliana wild type and the tlp18.3 mutant plants, the latter showing a stunted growth phenotype under fluctuating light conditions (Biochem. J, 406, 415-425. Expression pattern of genes encoding components of the photosynthetic electron transfer chain did not differ between fluctuating and constant light conditions, neither in wild type nor in tlp18.3 plants, and the composition of the thylakoid membrane protein complexes likewise remained unchanged. Nevertheless, the fluctuating light conditions repressed in wild-type plants a broad spectrum of genes involved in immune responses, which likely resulted from shade-avoidance responses and their intermixing with hormonal signaling. On the contrary, in the tlp18.3 mutant plants there was an imperfect repression of defense-related transcripts upon growth under fluctuating light, possibly by signals originating from minor malfunction of the photosystem II (PSII repair cycle, which directly or indirectly modulated the transcript abundances of genes related to light perception via phytochromes. Consequently, a strong allocation of resources to defense reactions in the tlp18.3 mutant plants presumably results in the stunted growth phenotype under fluctuating light.

Integrated membrane distillation-crystallization: process design and cost estimations for seawater treatment and fluxes of single salt solutions

NARCIS (Netherlands)

Creusen, R.J.M.; Medevoort, J. van; Roelands, C.P.M.; Renesse van Duivenbode, J.A.D. van; Hanemaaijer, J.H.; Leerdam, R.C. van

2013-01-01

The goal of this research is to design an integrated membrane distillation-crystallization (MDC) process for desalination of seawater with pure water and dry salts as the only products. The process is based on a combination of membrane distillation (MD) and osmotic distillation (OD) steps with

Stability of transmembrane amyloid β-peptide and membrane integrity tested by molecular modeling of site-specific Aβ42 mutations.

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Chetan Poojari

Full Text Available Interactions of the amyloid β-protein (Aβ with neuronal cell membranes, leading to the disruption of membrane integrity, are considered to play a key role in the development of Alzheimer's disease. Natural mutations in Aβ42, such as the Arctic mutation (E22G have been shown to increase Aβ42 aggregation and neurotoxicity, leading to the early-onset of Alzheimer's disease. A correlation between the propensity of Aβ42 to form protofibrils and its effect on neuronal dysfunction and degeneration has been established. Using rational mutagenesis of the Aβ42 peptide it was further revealed that the aggregation of different Aβ42 mutants in lipid membranes results in a variety of polymorphic aggregates in a mutation dependent manner. The mutant peptides also have a variable ability to disrupt bilayer integrity. To further test the connection between Aβ42 mutation and peptide-membrane interactions, we perform molecular dynamics simulations of membrane-inserted Aβ42 variants (wild-type and E22G, D23G, E22G/D23G, K16M/K28M and K16M/E22G/D23G/K28M mutants as β-sheet monomers and tetramers. The effects of charged residues on transmembrane Aβ42 stability and membrane integrity are analyzed at atomistic level. We observe an increased stability for the E22G Aβ42 peptide and a decreased stability for D23G compared to wild-type Aβ42, while D23G has the largest membrane-disruptive effect. These results support the experimental observation that the altered toxicity arising from mutations in Aβ is not only a result of the altered aggregation propensity, but also originates from modified Aβ interactions with neuronal membranes.

Formation of integral asymmetric membranes of AB diblock and ABC triblock copolymers by phase inversion.

Science.gov (United States)

Jung, Adina; Filiz, Volkan; Rangou, Sofia; Buhr, Kristian; Merten, Petra; Hahn, Janina; Clodt, Juliana; Abetz, Clarissa; Abetz, Volker

2013-04-12

The formation of integral asymmetric membranes from ABC triblock terpolymers by non-solvent-induced phase separation is shown. They are compared with the AB diblock copolymer precursors. Triblock terpolymers of polystyrene-block-poly(2-vinylpyridine)-block-poly(ethylene oxide) (PS-b-P2VP-b-PEO) with two compositions are investigated. The third block supports the formation of a membrane in a case, where the corresponding diblock copolymer does not form a good membrane. In addition, the hydrophilicity is increased by the third block and due to the hydroxyl group the possibility of post-functionalization is given. The morphologies are imaged by scanning electron microscopy. The influence of the PEO on the membrane properties is analyzed by water flux, retention, and dynamic contact angle measurements. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Electrolyte control of photosynthetic electron transport in cyanobacteria

International Nuclear Information System (INIS)

Papageorgiou, G.C.

1986-01-01

Ion-permeable cells (permeaplasts) of the cyanobacterium Anacystis nidulans were prepared enzymatically and were characterized with respect to several structural and functional indices. The permeaplasts contain intact, ion-impermeable thylakoids and are photosynthetically active. The authors discuss how, employing these cells, they investigated the effects of cations, acting either on the outer, or on the inner thylakoid membrane surface, on photoinduced electron exchanges with anionic donors (Cyt c-550, plastocyanin, innersurface), or anionic acceptors (FeCN 3- ; outer surface). Cations accelerate such exchanges by accumulating near the solution-membrane interfaces and screening the negative surface charge of membranes. Electrostatic screening, however, is not the only contributing factor, and other electrolyte-linked influences must be invoked in order to interpret the experimental observations

The 1.7 Å resolution structure of At2g44920, a pentapeptide-repeat protein in the thylakoid lumen of Arabidopsis thaliana

International Nuclear Information System (INIS)

Ni, Shuisong; McGookey, Michael E.; Tinch, Stuart L.; Jones, Alisha N.; Jayaraman, Seetharaman; Tong, Liang; Kennedy, Michael A.

2011-01-01

The crystal structure of At2g44920, a pentapeptide repeat protein (PRP) from Arabidopsis thaliana, has been determined at 1.7 Å resolution. The structure represents the first PRP protein whose subcellular localization has been experimentally confirmed to be the thylakoid lumen of a plant species. At2g44920 belongs to a diverse family (Pfam PF00805) of pentapeptide-repeat proteins (PRPs) that are present in all known organisms except yeast. PRPs contain at least eight tandem-repeating sequences of five amino acids with an approximate consensus sequence (STAV)(D/N)(L/F)(S/T/R)(X). Recent crystal structures show that PRPs adopt a highly regular four-sided right-handed β-helical structure consisting mainly of type II and type IV β-turns, sometimes referred to as a repeated five-residue (or Rfr) fold. Among sequenced genomes, PRP genes are most abundant in cyanobacteria, leading to speculation that PRPs play an important role in the unique lifestyle of photosynthetic cyanobacteria. Despite the recent structural characterization of several cyanobacterial PRPs, most of their functions remain unknown. Plants, whose chloroplasts are of cyanobacterial origin, have only four PRP genes in their genomes. At2g44920 is one of three PRPs located in the thylakoid lumen. Here, the crystal structure of a double methionine mutant of residues 81–224 of At2g44920, the naturally processed fragment of one of its full-length isoforms, is reported at 1.7 Å resolution. The structure of At2g44920 consists of the characteristic Rfr fold with five uninterrupted coils made up of 25 pentapeptide repeats and α-helical elements capping both termini. A disulfide bridge links the two α-helices with a conserved loop between the helical elements at its C-terminus. This structure represents the first structure of a PRP protein whose subcellular location has been experimentally confirmed to be the thylakoid lumen in a plant species

Molecular electronics of a single photosystem I reaction center: Studies with scanning tunneling microscopy and spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Lee, I.; Lee, J.W.; Warmack, R.J.; Allison, D.P.; Greenbaum, E. [Oak Ridge National Lab., TN (United States)

1995-03-14

Thylakoids and photosystem I (PSI) reaction centers were imaged by scanning tunneling microscopy. The thylakoids were isolated from spinach chloroplasts, and PSI reaction centers were extracted from thylakoid membranes. Because thylakoids are relatively thick nonconductors, they were sputter-coated with Pd/Au before imaging. PSI photosynthetic centers and chemically platinized PSI were investigated without sputter-coating. They were mounted on flat gold substrates that had been treated with mercaptoacetic acid to help bind the proteins. With tunneling spectroscopy, the PSI centers displayed a semiconductor-like response with a band gap of 1.8 eV. Lightly platinized (platinized for 1 hr) centers displayed diode-like conduction that resulted in dramatic contrast changes between images taken with opposite bias voltages. The electronic properties of this system were stable under long-term storage. 42 refs., 7 figs.

Evolutionary conservation of dual Sec translocases in the cyanelles of Cyanophora paradoxa

Directory of Open Access Journals (Sweden)

Löffelhardt Wolfgang

2008-11-01

Full Text Available Abstract Background Cyanelles, the peptidoglycan-armored plastids of glaucocystophytes, occupy a unique bridge position in between free-living cyanobacteria and chloroplasts. In some respects they side with cyanobacteria whereas other features are clearly shared with chloroplasts. The Sec translocase, an example for "conservative sorting" in the course of evolution, is found in the plasma membrane of all prokaryotes, in the thylakoid membrane of chloroplasts and in both these membrane types of cyanobacteria. Results In this paper we present evidence for a dual location of the Sec translocon in the thylakoid as well as inner envelope membranes of the cyanelles from Cyanophora paradoxa, i. e. conservative sorting sensu stricto. The prerequisite was the generation of specific antisera directed against cyanelle SecY that allowed immunodetection of the protein on SDS gels from both membrane types separated by sucrose density gradient floatation centrifugation. Immunoblotting of blue-native gels yielded positive but differential results for both the thylakoid and envelope Sec complexes, respectively. In addition, heterologous antisera directed against components of the Toc/Tic translocons and binding of a labeled precursor protein were used to discriminate between inner and outer envelope membranes. Conclusion The envelope translocase can be envisaged as a prokaryotic feature missing in higher plant chloroplasts but retained in cyanelles, likely for protein transport to the periplasm. Candidate passengers are cytochrome c6 and enzymes of peptidoglycan metabolism. The minimal set of subunits of the Toc/Tic translocase of a primitive plastid is proposed.

Spacesuit Water Membrane Evaporator Integration with the ISS Extravehicular Mobility

Science.gov (United States)

Margiott, Victoria; Boyle, Robert

2014-01-01

NASA has developed a Solid Water Membrane Evaporation (SWME) to provide cooling for the next generation spacesuit. One approach to increasing the TRL of the system is to incorporate this hardware with the existing EMU. Several integration issues were addressed to support a potential demonstration of the SWME with the existing EMU. Systems analysis was performed to assess the capability of the SWME to maintain crewmember cooling and comfort as a replacement for sublimation. The materials of the SWME were reviewed to address compatibility with the EMU. Conceptual system placement and integration with the EMU via an EVA umbilical system to ensure crew mobility and Airlock egress were performed. A concept of operation for EVA use was identified that is compatible with the existing system. This concept is extensible as a means to provide cooling for the existing EMU. The cooling system of one of the EMUs on orbit has degraded, with the root cause undetermined. Should there be a common cause resident on ISS, this integration could provide a means to recover cooling capability for EMUs on orbit.

The microalga Chlamydomonas reinhardtii CW-15 as a solar cell for hydrogen peroxide photoproduction. Comparison between free and immobilized cells and thylakoids for energy conversion efficiency

Energy Technology Data Exchange (ETDEWEB)

Scholz, W.; Galvan, F.; Rosa, F.F. de la [Instituto de Bioquimica Vegetal y Fotosintesis, Universidad de Sevilla y CSIC, Sevilla (Spain)

1995-11-28

Immobilized cells and thylakoid vesicles of the microalga Chlamydomonas reinhardtii CW-15 have been developed as a solar cell because of their capabilities of producing hydrogen peroxide. This compound is an efficient and clean fuel used for rocket propulsion, motors and for heating. Hydrogen peroxide is produced by the photosystem in a catalyst cycle in which a redox mediator (methyl viologen) is reduced by electrons obtained from water by the photosynthetic apparatus of the microalga and it is re-oxidized by the oxygen dissolved in the solution. The photoproduction has been investigated using a discontinuous system with whole cells, or thylakoid vesicles, free or immobilized on alginate. The stimulation by azide as an inhibitor of catalase has also been analyzed. Under determined optimum conditions, the photoproduction by Ca-alginate entrapped cells, with a rate of 33 {mu}mol H{sub 2}O{sub 2}/mg Chl.h, was maintained for several hours with an energy conversion efficiency of 0.25%

The short-term response of Arabidopsis thaliana (C3) and Zea mays (C4) chloroplasts to red and far red light.

Science.gov (United States)

Zienkiewicz, Maksymilian; Drożak, Anna; Wasilewska, Wioleta; Bacławska, Ilona; Przedpełska-Wąsowicz, Ewa; Romanowska, Elżbieta

2015-12-01

Light quality has various effects on photochemistry and protein phosphorylation in Zea mays and Arabidopsis thaliana thylakoids due to different degrees of light penetration across leaves and redox status in chloroplasts. The effect of the spectral quality of light (red, R and far red, FR) on the function of thylakoid proteins in Zea mays and Arabidopsis thaliana was investigated. It was concluded that red light stimulates PSII activity in A. thaliana thylakoids and in maize bundle sheath (BS) thylakoids, but not in mesophyll (M) thylakoids. The light quality did not change PSI activity in M thylakoids of maize. FR used after a white light period increased PSI activity significantly in maize BS and only slightly in A. thaliana thylakoids. As shown by blue native (BN)-PAGE followed by SDS-PAGE, proteins were differently phosphorylated in the thylakoids, indicating their different functions. FR light increased dephosphorylation of LHCII proteins in A. thaliana thylakoids, whereas in maize, dephosphorylation did not occur at all. The rate of phosphorylation was higher in maize BS than in M thylakoids. D1 protein phosphorylation increased in maize and decreased in A. thaliana upon irradiation with both R and growth light (white light, W). Light variations did not change the level of proteins in thylakoids. Our data strongly suggest that response to light quality is a species-dependent phenomenon. We concluded that the maize chloroplasts were differently stimulated, probably due to different degrees of light penetration across the leaf and thereby the redox status in the chloroplasts. These acclimation changes induced by light quality are important in the regulation of chloroplast membrane flexibility and thus its function.

Integrating Microbial Electrochemical Technology with Forward Osmosis and Membrane Bioreactors: Low-Energy Wastewater Treatment, Energy Recovery and Water Reuse

KAUST Repository

Werner, Craig M.

2014-06-01

Wastewater treatment is energy intensive, with modern wastewater treatment processes consuming 0.6 kWh/m3 of water treated, half of which is required for aeration. Considering that wastewater contains approximately 2 kWh/m3 of energy and represents a reliable alternative water resource, capturing part of this energy and reclaiming the water would offset or even eliminate energy requirements for wastewater treatment and provide a means to augment traditional water supplies. Microbial electrochemical technology is a novel technology platform that uses bacteria capable of producing an electric current outside of the cell to recover energy from wastewater. These bacteria do not require oxygen to respire but instead use an insoluble electrode as their terminal electron acceptor. Two types of microbial electrochemical technologies were investigated in this dissertation: 1) a microbial fuel cell that produces electricity; and 2) a microbial electrolysis cell that produces hydrogen with the addition of external power. On their own, microbial electrochemical technologies do not achieve sufficiently high treatment levels. Innovative approaches that integrate microbial electrochemical technologies with emerging and established membrane-based treatment processes may improve the overall extent of wastewater treatment and reclaim treated water. Forward osmosis is an emerging low-energy membrane-based technology for seawater desalination. In forward osmosis water is transported across a semipermeable membrane driven by an osmotic gradient. The microbial osmotic fuel cell described in this dissertation integrates a microbial fuel cell with forward osmosis to achieve wastewater treatment, energy recovery and partial desalination. This system required no aeration and generated more power than conventional microbial fuel cells using ion exchange membranes by minimizing electrochemical losses. Membrane bioreactors incorporate semipermeable membranes within a biological wastewater

Membrane composition and physiological activity of plastids from an oenothera plastome mutator-induced chloroplast mutant.

Science.gov (United States)

Johnson, E M; Sears, B B

1990-01-01

Plastids were isolated from a plastome mutator-induced mutant (pm7) of Oenothera hookeri and were analyzed for various physiological and biochemical attributes. No photosynthetic electron transport activity was detected in the mutant plastids. This is consistent with previous ultrastructural analysis showing the absence of thylakoid membranes in the pm7 plastids and with the observation of aberrant processing and accumulation of chloroplast proteins in the mutant. In comparison to wild type, the mutant tissue lacks chlorophyll, and has significant differences in levels of four fatty acids. The analyses did not reveal any differences in carotenoid levels nor in the synthesis of several chloroplast lipids. The consequences of the altered composition of the chloroplast membrane are discussed in terms of their relation to the aberrant protein processing of the pm7 plastids. The pigment, fatty acid, and lipid measurements were also performed on two distinct nuclear genotypes (A/A and A/C) which differ in their compatibility with the plastid genome (type I) contained in these lines. In these cases, only chlorophyll concentrations differed significantly.

Differential Mobility of Pigment-Protein Complexes in Granal and Agranal Thylakoid Membranes of C-3 and C-4 Plants

Czech Academy of Sciences Publication Activity Database

Kirchhoff, H.; Sharpe, R.M.; Herbstová, Miroslava; Yarbrough, R.; Edwards, G.E.

2013-01-01

Roč. 161, č. 1 (2013), s. 497-507 ISSN 0032-0889 Institutional support: RVO:60077344 Keywords : Photosystem-II * Photosynthetic membranes * Electron tomography Subject RIV: ED - Physiology Impact factor: 7.394, year: 2013

Hybrid bio-photo-electro-chemical cells for solar water splitting.

Science.gov (United States)

Pinhassi, Roy I; Kallmann, Dan; Saper, Gadiel; Dotan, Hen; Linkov, Artyom; Kay, Asaf; Liveanu, Varda; Schuster, Gadi; Adir, Noam; Rothschild, Avner

2016-08-23

Photoelectrochemical water splitting uses solar power to decompose water to hydrogen and oxygen. Here we show how the photocatalytic activity of thylakoid membranes leads to overall water splitting in a bio-photo-electro-chemical (BPEC) cell via a simple process. Thylakoids extracted from spinach are introduced into a BPEC cell containing buffer solution with ferricyanide. Upon solar-simulated illumination, water oxidation takes place and electrons are shuttled by the ferri/ferrocyanide redox couple from the thylakoids to a transparent electrode serving as the anode, yielding a photocurrent density of 0.5 mA cm(-2). Hydrogen evolution occurs at the cathode at a bias as low as 0.8 V. A tandem cell comprising the BPEC cell and a Si photovoltaic module achieves overall water splitting with solar to hydrogen efficiency of 0.3%. These results demonstrate the promise of combining natural photosynthetic membranes and man-made photovoltaic cells in order to convert solar power into hydrogen fuel.

Effect of commercial long-term extenders on metabolic activity and membrane integrity of boar spermatozoa stored at 17 degrees C.

Science.gov (United States)

Dziekońska, A; Fraser, L; Majewska, A; Lecewicz, M; Zasiadczyk, Ł; Kordan, W

2013-01-01

This study was aimed to analyze the metabolic activity and membrane integrity of boar spermatozoa following storage in long-term semen extenders. Boar semen was diluted with Androhep EnduraGuard (AeG), DILU-Cell (DC), SafeCell Plus (SCP) and Vitasem LD (VLD) extenders and stored for 10 days at 17 degrees C. Parameters of the analyzed sperm metabolic activity included total motility (TMOT), progressive motility (PMOT), high mitochondrial membrane potential (MMP) and ATP content, whereas those of the membrane integrity included plasma membrane integrity (PMI) and normal apical ridge (NAR) acrosome. Extender type was a significant (P semen storage. In all extenders the metabolic activity and membrane integrity of the stored spermatozoa decreased continuously over time. Among the four analyzed extenders, AeG and SCP showed the best performance in terms of TMOT and PMI on Days 5, 7 and 10 of storage. Marked differences in the proportions of spermatozoa with high MMP were observed between the extenders, particularly on Day 10 of storage. There were not any marked differences in sperm ATP content between the extenders, regardless of the storage time. Furthermore, the percentage of spermatozoa with NAR acrosomes decreased during prolonged storage, being markedly lower in DC-diluted semen compared with semen diluted with either AeG or SCP extender. The results of this study indicated that components of the long-term extenders have different effects on the sperm functionality and prolonged semen longevity by delaying the processes associated with sperm ageing during liquid storage.

Self-organizing amyloid in bacteria

DEFF Research Database (Denmark)

Otzen, D. E.; Vad, B. S.; Dueholm, M. S.

2017-01-01

Real biological membranes are complex multi-component systems which are not easily subjected to systematic investigations, although e.g. the dynamics of light-induced structural reorganizations of thylakoid membranes in chloroplast organelles have been studied using time-resolved small-angle neut...

Catalytic, Conductive Bipolar Membrane Interfaces through Layer-by-Layer Deposition for the Design of Membrane-Integrated Artificial Photosynthesis Systems.

Science.gov (United States)

McDonald, Michael B; Freund, Michael S; Hammond, Paula T

2017-11-23

In the presence of an electric field, bipolar membranes (BPMs) are capable of initiating water disassociation (WD) within the interfacial region, which can make water splitting for renewable energy in the presence of a pH gradient possible. In addition to WD catalytic efficiency, there is also the need for electronic conductivity in this region for membrane-integrated artificial photosynthesis (AP) systems. Graphene oxide (GO) was shown to catalyze WD and to be controllably reduced, which resulted in electronic conductivity. Layer-by-layer (LbL) film deposition was employed to improve GO film uniformity in the interfacial region to enhance WD catalysis and, through the addition of a conducting polymer in the process, add electronic conductivity in a hybrid film. Three different deposition methods were tested to optimize conducting polymer synthesis with the oxidant in a metastable solution and to yield the best film properties. It was found that an approach that included substrate dipping in a solution containing the expected final monomer/oxidant ratio provided the most predictable film growth and smoothest films (by UV/Vis spectroscopy and atomic force microscopy/scanning electron microscopy, respectively), whereas dipping in excess oxidant or co-spraying the oxidant and monomer produced heterogeneous films. Optimized films were found to be electronically conductive and produced a membrane ohmic drop that was acceptable for AP applications. Films were integrated into the interfacial region of BPMs and revealed superior WD efficiency (≥1.4 V at 10 mA cm -2 ) for thinner films (<10 bilayers≈100 nm) than for either the pure GO catalyst or conducting polymer individually, which indicated that there was a synergistic effect between these materials in the structure configured by the LbL method. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Integral and peripheral association of proteins and protein complexes with Yersinia pestis inner and outer membranes

Directory of Open Access Journals (Sweden)

Bunai Christine L

2009-02-01

Full Text Available Abstract Yersinia pestis proteins were sequentially extracted from crude membranes with a high salt buffer (2.5 M NaBr, an alkaline solution (180 mM Na2CO3, pH 11.3 and membrane denaturants (8 M urea, 2 M thiourea and 1% amidosulfobetaine-14. Separation of proteins by 2D gel electrophoresis was followed by identification of more than 600 gene products by MS. Data from differential 2D gel display experiments, comparing protein abundances in cytoplasmic, periplasmic and all three membrane fractions, were used to assign proteins found in the membrane fractions to three protein categories: (i integral membrane proteins and peripheral membrane proteins with low solubility in aqueous solutions (220 entries; (ii peripheral membrane proteins with moderate to high solubility in aqueous solutions (127 entries; (iii cytoplasmic or ribosomal membrane-contaminating proteins (80 entries. Thirty-one proteins were experimentally associated with the outer membrane (OM. Circa 50 proteins thought to be part of membrane-localized, multi-subunit complexes were identified in high Mr fractions of membrane extracts via size exclusion chromatography. This data supported biologically meaningful assignments of many proteins to the membrane periphery. Since only 32 inner membrane (IM proteins with two or more predicted transmembrane domains (TMDs were profiled in 2D gels, we resorted to a proteomic analysis by 2D-LC-MS/MS. Ninety-four additional IM proteins with two or more TMDs were identified. The total number of proteins associated with Y. pestis membranes increased to 456 and included representatives of all six β-barrel OM protein families and 25 distinct IM transporter families.

Reference: 483 [Arabidopsis Phenome Database[Archive

Lifescience Database Archive (English)

Full Text Available ent and protein content of the thylakoid membranes were otherwise almost unchanged. However, there was a maj...or change in the macroorganization of PSII within these membranes; electron micro...ing complex CP24 affects the structure and function of the grana membranes of higher plant chloroplasts. 11

Very Low Surface Energy (Membrane Separations: An Integrated Polymer Chemistry/Engineering Approach and The Influence of Backpulsing on Fouling Properties of Novel Nanofiltration Membranes for Wastewater Remediation

National Research Council Canada - National Science Library

Freeman, Benny

1998-01-01

...: An Integrated Polymer Chemistry/Engineering Approach, is to explore several new classes of polymeric materials to identify promising routes for developing low-fouling nanofiltration membranes for wastewater remediation...

Effects of alpha-lipoic acids on sperm membrane integrity during liquid storage of boar semen

Directory of Open Access Journals (Sweden)

Laura Parlapan

2015-05-01

Full Text Available Preliminary studies have shown that sperm membrane from swine shows high sensitivity to cryopreservation process, causing a dramatic reduction in sperm quality. This has been attributed to the production of reactive oxygen species, that cause lipid peroxidation in sperm membranes. The aim of the present study was to minimize the oxidative attack by adding different concentration of alpha-lipoic acid into the sperm liquid storage at 17ºC for 7 days. Freshly ejaculated boar semen was diluted with Beltsville Thawing Solution (BTS and supplemented with 5 levels of alpha-lipoic  acid (0.015, 0.02, 0.05, 0.1, 0.15 mmol/ml. The membrane integrity was evaluated at days 0, 1, 3, 5 and 7 of liquid preservation, using flow cytometer FACSCanto II (BD Biociencias systems. The experiment indicate that supplementation of alpha-lipoic  acid to the semen liquid storage extender improve sperm membrane

Integration of ceramic membrane and compressed air-assisted solvent extraction (CASX) for metal recovery.

Science.gov (United States)

Li, Chi-Wang; Chiu, Chun-Hao; Lee, Yu-Cheng; Chang, Chia-Hao; Lee, Yu-Hsun; Chen, Yi-Ming

2010-01-01

In our previous publications, compressed air-assisted solvent extraction process (CASX) was developed and proved to be kinetically efficient process for metal removal. In the current study, CASX with a ceramic MF membrane integrated for separation of spent solvent was employed to remove and recover metal from wastewater. MF was operated either in crossflow mode or dead-end with intermittent flushing mode. Under crossflow mode, three distinct stages of flux vs. TMP (trans-membrane pressure) relationship were observed. In the first stage, flux increases with increasing TMP which is followed by the stage of stable flux with increasing TMP. After reaching a threshold TMP which is dependent of crossflow velocity, flux increases again with increasing TMP. At the last stage, solvent was pushed through membrane pores as indicated by increasing permeate COD. In dead-end with intermittent flushing mode, an intermittent flushing flow (2 min after a 10-min or a 30-min dead-end filtration) was incorporated to reduce membrane fouling by flush out MSAB accumulated on membrane surface. Effects of solvent concentration and composition were also investigated. Solvent concentrations ranging from 0.1 to 1% (w/w) have no adverse effect in terms of membrane fouling. However, solvent composition, i.e. D(2)EHPA/kerosene ratio, shows impact on membrane fouling. The type of metal extractants employed in CASX has significant impact on both membrane fouling and the quality of filtrate due to the differences in their viscosity and water solubility. Separation of MSAB was the limiting process controlling metal removal efficiency, and the removal efficiency of Cd(II) and Cr(VI) followed the same trend as that for COD.

Direct effects of ionizing radiation on integral membrane proteins. Noncovalent energy transfer requires specific interpeptide interactions

International Nuclear Information System (INIS)

Jhun, E.; Jhun, B.H.; Jones, L.R.; Jung, C.Y.

1991-01-01

The 12 transmembrane alpha helices (TMHs) of human erythrocyte glucose transporter were individually cut by pepsin digestion as membrane-bound 2.5-3.5-kDa peptide fragments. Radiation-induced chemical degradation of these fragments showed an average target size of 34 kDa. This is 10-12 x larger than the average size of an individual TMH, demonstrating that a significant energy transfer occurs among these TMHs in the absence of covalent linkage. Heating this TMH preparation at 100 degree C for 15 min reduced the target size to 5 kDa or less, suggesting that the noncovalent energy transfer requires specific helix-helix interactions. Purified phospholamban, a small (6-kDa) integral membrane protein containing a single TMH, formed a pentameric assembly in sodium dodecyl sulfate. The chemical degradation target size of this phospholamban pentamer was 5-6 kDa, illustrating that not all integral membrane protein assemblies permit intersubunit energy transfer. These findings together with other published observations suggest strongly that significant noncovalent energy transfer can occur within the tertiary and quaternary structure of membrane proteins and that as yet undefined proper molecular interactions are required for such covalent energy transfer. Our results with pepsin-digested glucose transporter also illustrate the importance of the interhelical interaction as a predominating force in maintaining the tertiary structure of a transmembrane protein

Hemorrhagic shock impairs myocardial cell volume regulation and membrane integrity in dogs

International Nuclear Information System (INIS)

Horton, J.W.

1987-01-01

An in vitro myocardial slice technique was used to quantitate alterations in cell volume regulation and membrane integrity after 2 h or hemorrhagic shock. After in vitro incubation in Krebs-Ringer-phosphate medium containing trace [ 14 C]inulin, values (ml H 2 O/g dry wt) for control nonshocked myocardial slices were 4.03 /plus minus/ 0.11 (SE) for total water, 2.16 /plus minus/ 0.07 for inulin impermeable space, and 1.76 /plus minus/ 0.15 for inulin diffusible space. Shocked myocardial slices showed impaired response to cold incubation. After 2 h of in vivo shock, total tissue water, inulin diffusible space, and inulin impermeable space increased significantly for subendocardium, whereas changes in subepicardium parameters were minimal. Shock-induced cellular swelling was accompanied by an increased total tissue sodium, but no change in tissue potassium. Calcium entry blockade in vivo significantly reduced subendocardial total tissue water as compared with shock-untreated dogs. In addition, calcium entry blockade reduced shock-induced increases in inulin diffusible space. In vitro myocardial slice studies confirm alterations in subendocardial membrane integrity after 2 h of in vivo hemorrhagic shock. Shock-induced abnormalities in myocardial cell volume regulation are reduced by calcium entry blockade in vivo

Lipid bilayer-bound conformation of an integral membrane beta barrel protein by multidimensional MAS NMR

International Nuclear Information System (INIS)

Eddy, Matthew T.; Su, Yongchao; Silvers, Robert; Andreas, Loren; Clark, Lindsay; Wagner, Gerhard; Pintacuda, Guido; Emsley, Lyndon; Griffin, Robert G.

2015-01-01

The human voltage dependent anion channel 1 (VDAC) is a 32 kDa β-barrel integral membrane protein that controls the transport of ions across the outer mitochondrial membrane. Despite the determination of VDAC solution and diffraction structures, a structural basis for the mechanism of its function is not yet fully understood. Biophysical studies suggest VDAC requires a lipid bilayer to achieve full function, motivating the need for atomic resolution structural information of VDAC in a membrane environment. Here we report an essential step toward that goal: extensive assignments of backbone and side chain resonances for VDAC in DMPC lipid bilayers via magic angle spinning nuclear magnetic resonance (MAS NMR). VDAC reconstituted into DMPC lipid bilayers spontaneously forms two-dimensional lipid crystals, showing remarkable spectral resolution (0.5–0.3 ppm for 13 C line widths and <0.5 ppm 15 N line widths at 750 MHz). In addition to the benefits of working in a lipid bilayer, several distinct advantages are observed with the lipid crystalline preparation. First, the strong signals and sharp line widths facilitated extensive NMR resonance assignments for an integral membrane β-barrel protein in lipid bilayers by MAS NMR. Second, a large number of residues in loop regions were readily observed and assigned, which can be challenging in detergent-solubilized membrane proteins where loop regions are often not detected due to line broadening from conformational exchange. Third, complete backbone and side chain chemical shift assignments could be obtained for the first 25 residues, which comprise the functionally important N-terminus. The reported assignments allow us to compare predicted torsion angles for VDAC prepared in DMPC 2D lipid crystals, DMPC liposomes, and LDAO-solubilized samples to address the possible effects of the membrane mimetic environment on the conformation of the protein. Concluding, we discuss the strengths and weaknesses of the reported

Antibacterial Effect of Gallic Acid against Aeromonas hydrophila and Aeromonas sobria Through Damaging Membrane Integrity.

Science.gov (United States)

Lu, Jing; Wang, Zhenning; Ren, Mengrou; Huang, Guoren; Fang, Baochen; Bu, Xiujuan; Liu, Yanhui; Guan, Shuang

In the study, we investigated the antibacterial activity and mechanism of gallic acid against Aeromonas hydrophila and Aeromonas sobria. Gallic acid showed strong antimicrobial activity against the two bacteria. Furthermore, the antibacterial mechanism of gallic acid (0, 3, 6, 12 mM) was performed by membrane integrity assay and scanning electron microscopy (SEM) assay. The results showed that gallic acid notably increased the released material absorption value at 260, 280 nm and electric conductivity in a dose-dependent manner. Moreover, the SEM assay showed that gallic acid induced severe shrink of bacterial intima and irregular morphology in a dose-dependent manner. The SDS-PAGE profiles further confirmed that gallic acid could damage bacterial cells. These results indicated gallic acid exhibited antibacterial effect by destroying membrane integrity of A. hydrophila and A. sobria. Hence, gallic acid has great potential as a new natural food preservative in food fresh-keeping and storage.

Critical review of membrane bioreactor models--part 2: hydrodynamic and integrated models.

Science.gov (United States)

Naessens, W; Maere, T; Ratkovich, N; Vedantam, S; Nopens, I

2012-10-01

Membrane bioreactor technology exists for a couple of decades, but has not yet overwhelmed the market due to some serious drawbacks of which operational cost due to fouling is the major contributor. Knowledge buildup and optimisation for such complex systems can heavily benefit from mathematical modelling. In this paper, the vast literature on hydrodynamic and integrated MBR modelling is critically reviewed. Hydrodynamic models are used at different scales and focus mainly on fouling and only little on system design/optimisation. Integrated models also focus on fouling although the ones including costs are leaning towards optimisation. Trends are discussed, knowledge gaps identified and interesting routes for further research suggested. Copyright © 2012 Elsevier Ltd. All rights reserved.

Integration of mixed conducting membranes in an oxygen–steam biomass gasification process

DEFF Research Database (Denmark)

Puig Arnavat, Maria; Soprani, Stefano; Søgaard, Martin

2013-01-01

. The two configurations demonstrating the highest efficiency are then thermally integrated into an oxygen– steam biomass gasification plant. The energy demand for oxygen production and the membrane area required for a 6 MWth biomass plant are calculated for different operating conditions. Increasing......Oxygen–steam biomass gasification produces a high quality syngas with a high H2/CO ratio that is suitable for upgrading to liquid fuels. Such a gas is also well suited for use in conjunction with solid oxide fuel cells giving rise to a system yielding high electrical efficiency based on biomass...... distillation, especially for small to medium scale plants. This paper examines different configurations for oxygen production using MIEC membranes where the oxygen partial pressure difference is achieved by creating a vacuum on the permeate side, compressing the air on the feed side or a combination of the two...

Membrane Composition and Physiological Activity of Plastids from an Oenothera Plastome Mutator-Induced Chloroplast Mutant 1

Science.gov (United States)

Johnson, Ellen M.; Sears, Barbara B.

1990-01-01

Plastids were isolated from a plastome mutator-induced mutant (pm7) of Oenothera hookeri and were analyzed for various physiological and biochemical attributes. No photosynthetic electron transport activity was detected in the mutant plastids. This is consistent with previous ultrastructural analysis showing the absence of thylakoid membranes in the pm7 plastids and with the observation of aberrant processing and accumulation of chloroplast proteins in the mutant. In comparison to wild type, the mutant tissue lacks chlorophyll, and has significant differences in levels of four fatty acids. The analyses did not reveal any differences in carotenoid levels nor in the synthesis of several chloroplast lipids. The consequences of the altered composition of the chloroplast membrane are discussed in terms of their relation to the aberrant protein processing of the pm7 plastids. The pigment, fatty acid, and lipid measurements were also performed on two distinct nuclear genotypes (A/A and A/C) which differ in their compatibility with the plastid genome (type I) contained in these lines. In these cases, only chlorophyll concentrations differed significantly. PMID:16667256

Insulin alters the target size of the peripheral cyclic AMP phosphodiesterase but not the integral cyclic GMP-stimulated cyclic AMP phosphodiesterase in liver plasma membranes

International Nuclear Information System (INIS)

Wallace, A.V.; Martin, B.R.; Houslay, M.D.

1990-01-01

Radiation inactivation of the two high affinity cyclic AMP phosphodiesterases (PDE) found in liver plasma membranes afforded an estimation of their molecular target sizes in situ. The activity of the peripheral plasma membrane PDE decayed as a single exponential with a target size corresponding to a monomer of circa 54 kDa. The integral, cyclic GMP-stimulated PDE decayed as a dimer of circa 125 kDa. Preincubation of plasma membranes with insulin (10nM), prior to irradiation, caused the target size of only the peripheral plasma membrane PDE to increase. We suggest that insulin addition causes the peripheral plasma membrane PDE to alter its coupling to an integral plasma membrane protein with a target size of circa 90 kDa

Expression of the recombinant bacterial outer surface protein A in tobacco chloroplasts leads to thylakoid localization and loss of photosynthesis.

Science.gov (United States)

Hennig, Anna; Bonfig, Katharina; Roitsch, Thomas; Warzecha, Heribert

2007-11-01

Bacterial lipoproteins play crucial roles in host-pathogen interactions and pathogenesis and are important targets for the immune system. A prominent example is the outer surface protein A (OspA) of Borrelia burgdorferi, which has been efficiently used as a vaccine for the prevention of Lyme disease. In a previous study, OspA could be produced in tobacco chloroplasts in a lipidated and immunogenic form. To further explore the potential of chloroplasts for the production of bacterial lipoproteins, the role of the N-terminal leader sequence was investigated. The amount of recombinant OspA could be increased up to ten-fold by the variation of the insertion site in the chloroplast genome. Analysis of OspA mutants revealed that replacement of the invariant cysteine residue as well as deletion of the leader sequence abolishes palmitolyation of OspA. Also, decoration of OspA with an N-terminal eukaryotic lipidation motif does not lead to palmitoylation in chloroplasts. Strikingly, the bacterial signal peptide of OspA efficiently targets the protein to thylakoids, and causes a mutant phenotype. Plants accumulating OspA at 10% total soluble protein could not grow without exogenously supplied sugars and rapidly died after transfer to soil under greenhouse conditions. The plants were found to be strongly affected in photosystem II, as revealed by the analyses of temporal and spatial dynamics of photosynthetic activity by chlorophyll fluorescence imaging. Thus, overexpression of OspA in chloroplasts is limited by its concentration-dependent interference with essential functions of chloroplastic membranes required for primary metabolism.

Hybrid bio-photo-electro-chemical cells for solar water splitting

OpenAIRE

Pinhassi, Roy I.; Kallmann, Dan; Saper, Gadiel; Dotan, Hen; Linkov, Artyom; Kay, Asaf; Liveanu, Varda; Schuster, Gadi; Adir, Noam; Rothschild, Avner

2016-01-01

Photoelectrochemical water splitting uses solar power to decompose water to hydrogen and oxygen. Here we show how the photocatalytic activity of thylakoid membranes leads to overall water splitting in a bio-photo-electro-chemical (BPEC) cell via a simple process. Thylakoids extracted from spinach are introduced into a BPEC cell containing buffer solution with ferricyanide. Upon solar-simulated illumination, water oxidation takes place and electrons are shuttled by the ferri/ferrocyanide redox...

Ethanol production in an integrated fermentation/membrane system. Process simulations and economics

Energy Technology Data Exchange (ETDEWEB)

Groot, W J; Kraayenbrink, M R; Lans, R.G.J.M. van der; Luyben, K C.A.M. [Delft Univ. of Technology (Netherlands). Dept. of Biochemical Engineering

1993-01-01

Four systems comprising of an ethanol fermentation integrated with microfiltration and/or pervaporation, and a conventional continuous culture, were compared with respect to the performance of the fermentation and economics. The processes are compared on the basis of the same kinetic model. It is found that cell retention by microfiltration leads to lower production costs, compared to a conventional continuous culture. Pervaporation becomes profitable at a high selectivity of ethanol/water separation and low membrane prices. (orig.).

Single-Molecule Studies of Bacterial Protein Translocation

NARCIS (Netherlands)

Kedrov, Alexej; Kusters, Ilja; Driessen, Arnold J. M.

2013-01-01

In prokaryotes, a large share of the proteins are secreted from the cell through a process that requires their translocation across the cytoplasmic membrane. This process is mediated by the universally conserved Sec system with homologues in the endoplasmic reticulum and thylakoid membranes of

Desiccation induces accumulations of antheraxanthin and zeaxanthin in intertidal macro-alga Ulva pertusa (Chlorophyta.

Directory of Open Access Journals (Sweden)

Xiujun Xie

Full Text Available For plants and algae, exposure to high light levels is deleterious to their photosynthetic machineries. It also can accelerate water evaporation and thus potentially lead to drought stress. Most photosynthetic organisms protect themselves against high light caused photodamages by xanthophyll cycle-dependent thermal energy dissipation. It is generally accepted that high light activates xanthophyll cycle. However, the relationship between xanthophyll cycle and drought stress remains ambiguous. Herein, Ulva pertusa (Chlorophyta, a representative perennial intertidal macro-algae species with high drought-tolerant capabilities and simple structures, was used to investigate the operation of xanthophyll cycle during desiccation in air. The results indicate that desiccation under dim light induced accumulation of antheraxanthin (Ax and zeaxanthin (Zx at the expense of violaxanthin (Vx. This accumulation could be arrested by dithiothreitol completely and by uncoupler (carbonyl cyanide p-trifluoromethoxyphenylhydrazone partially, implying the participation of Vx de-epoxidase in conversion of Vx to Ax and Zx. Treatment with inhibitors of electron transport along thylakoid membrane, e.g. DCMU, PG and DBMIB, did not significantly arrest desiccation-induced accumulation of Ax and Zx. We propose that for U. pertusa, besides excess light, desiccation itself could also induce accumulation of Ax and Zx. This accumulation could proceed without electron transport along thylakoid membrane, and is possibly resulting from the reduction of thylakoid lumen volume during desiccation. Considering the pleiotropic effects of Ax and Zx, accumulated Ax and Zx may function in protecting thylakoid membrane and enhancing thermal quenching during emersion in air.

Effect of storage in short--and long-term commercial semen extenders on the motility, plasma membrane and chromatin integrity of boar spermatozoa.

Science.gov (United States)

De Ambrogi, Marco; Ballester, Juan; Saravia, Fernando; Caballero, Ignacio; Johannisson, Anders; Wallgren, Margareta; Andersson, Magnus; Rodriguez-Martinez, Heriberto

2006-10-01

For artificial insemination (AI) in pigs, preservation of liquid boar semen at 16-20 degrees C is still common practice as sperm cryopreservation remains suboptimal in this species. To meet the different needs of the swine industry, several extenders have been developed to preserve semen in liquid form for short--and long-term storage. In the present study, three different commercial extenders devised for short-term (BTS+) or long-term preservation (MR-A and X-Cell), were used to test whether storage of semen from four mature, fertile boars at 17 degrees C for 96 h would affect sperm characteristics relevant for fertility, such as motility, membrane integrity and chromatin stability. Computer-assisted sperm analysis, and stainings with the acylated membrane dye SYBR-14/propidium iodide, and acridine orange in connection with flow cytometry were used to evaluate these variables. Percentages of total motile spermatozoa decreased slightly, but significantly, after 72-96 h. While membrane integrity values varied during the period of study, no significant changes in either membrane integrity or chromatin stability were, however, registered. This suggests a customary 96-day storage at 17 degrees C in these extenders was too short an interval to cause losses of integrity in nuclear DNA in the boar population studied.

Metabolic dysfunction and unabated respiration precede the loss of membrane integrity during dehydration of germinating radicles

NARCIS (Netherlands)

Leprince, O.; Harren, F.J.M.; Alberda, M.; Hoekstra, F.A.

2000-01-01

This study shows that dehydration induces imbalanced metabolism before loss of membrane integrity in desiccation-sensitive germinated radicles. Using a photoacoustic detection system, responses of CO2 emission and fermentation to drying were analyzed non-invasively in desiccation-tolerant and

[THE EFFECT OF ACID RAIN ON ULTRASTRUCTURE AND FUNCTIONAL PARAMETERS OF PHOTOSYNTHETIC APPARATUS OF PEA LEAVES].

Science.gov (United States)

Polishchuk, A V; Vodka, M V; Belyavskaya, N A; Khomochkin, A P; Zolotareva, E K

2016-01-01

The effects of simulated acid rain (SAR) on the ultrastructure and functional parameters of the photosynthetic apparatus were studied using 14-day-old pea leaves as test system. Pea plants were sprayed with an aqueous solution containing NaNO₃(0.2 mM) and Na₂SO₄(0.2 mM) (pH 5.6, a control variant), or with the same solution, which was acidified to pH 2.5 (acid variant). Functional characteristics were determined by chlorophyll fluorescence analysis. Acid rain application caused reduction in the efficiency of the photosynthetic electron transport by 25%, which was accompanied by an increase by 85% in the quantum yield of thermal dissipation of excess light quanta. Ultrastructural changes in chloroplast were registered by transmission electron microscopy (TEM) after two days of the SAR-treatment of pea leaves. In this case, the changes in the structure of grana, heterogeneity of thylakoids packaging in granum, namely, the increase of intra-thylakoid gaps and thickness of granal thylakoids compared to the control were found. The migration of protein complexes in thylakoid membranes of chloroplasts isolated from leaves treated with SAR was suppressed. It was shown also that carbonic anhydrase activity was inhibited in chloroplast preparations isolated from SAR-treated pea leaves. We proposed a hypothesis on the possible inactivation of thylakoid carbonic anhydrase under SAR and its involvement in the inhibition of photochemical activity of chloroplasts. The data obtained allows to suggest that acid rains negatively affect the photosynthetic apparatus disrupting the membrane system of chloroplast.

Isolation of plant Photosystem II complexes by fractional solubilization

Directory of Open Access Journals (Sweden)

Patrycja eHaniewicz

2015-12-01

Full Text Available PSII occurs in different forms and supercomplexes in thylakoid membranes. Using a transplastomic strain of Nicotiana tabacum histidine tagged on the subunit PsbE, we have previously shown that a mild extraction protocol with β-dodecylmaltoside enriches PSII characteristic of lamellae and grana margins. Here, we characterize residual granal PSII that is not extracted by this first solubilization step. Using affinity purification, we demonstrate that this PSII fraction consists of PSII-LHCII mega- and supercomplexes, PSII dimers and PSII monomers, which were separated by gel filtration and functionally characterized. Our findings represent an alternative demonstration of different PSII populations in thylakoid membranes, and they make it possible to prepare PSII-LHCII supercomplexes in high yield.

Integrated pyrolucite fluidized bed-membrane hybrid process for improved iron and manganese control in drinking water.

Science.gov (United States)

Dashtban Kenari, Seyedeh Laleh; Barbeau, Benoit

2017-04-15

Newly developed ceramic membrane technologies offer numerous advantages over the conventional polymeric membranes. This work proposes a new configuration, an integrated pyrolucite fluidized bed (PFB)-ceramic MF/UF hybrid process, for improved iron and manganese control in drinking water. A pilot-scale study was undertaken to evaluate the performance of this process with respect to iron and manganese control as well as membrane fouling. In addition, the fouling of commercially available ceramic membranes in conventional preoxidation-MF/UF process was compared with the hybrid process configuration. In this regard, a series of experiments were conducted under different influent water quality and operating conditions. Fouling mechanisms and reversibility were analyzed using blocking law and resistance-in-series models. The results evidenced that the flux rate and the concentration of calcium and humic acids in the feed water have a substantial impact on the filtration behavior of both membranes. The model for constant flux compressible cake formation well described the rise in transmembrane pressure. The compressibility of the filter cake substantially increased in the presence of 2 mg/L humic acids. The presence of calcium ions caused significant aggregation of manganese dioxide and humic acid which severely impacted the extent of membrane fouling. The PFB pretreatment properly alleviated membrane fouling by removing more than 75% and 95% of iron and manganese, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

Profiling of integral membrane proteins and their post translational modifications using high-resolution mass spectrometry

Science.gov (United States)

Souda, Puneet; Ryan, Christopher M.; Cramer, William A.; Whitelegge, Julian

2011-01-01

Integral membrane proteins pose challenges to traditional proteomics approaches due to unique physicochemical properties including hydrophobic transmembrane domains that limit solubility in aqueous solvents. A well resolved intact protein molecular mass profile defines a protein’s native covalent state including post-translational modifications, and is thus a vital measurement toward full structure determination. Both soluble loop regions and transmembrane regions potentially contain post-translational modifications that must be characterized if the covalent primary structure of a membrane protein is to be defined. This goal has been achieved using electrospray-ionization mass spectrometry (ESI-MS) with low-resolution mass analyzers for intact protein profiling, and high-resolution instruments for top-down experiments, toward complete covalent primary structure information. In top-down, the intact protein profile is supplemented by gas-phase fragmentation of the intact protein, including its transmembrane regions, using collisionally activated and/or electroncapture dissociation (CAD/ECD) to yield sequence-dependent high-resolution MS information. Dedicated liquid chromatography systems with aqueous/organic solvent mixtures were developed allowing us to demonstrate that polytopic integral membrane proteins are amenable to ESI-MS analysis, including top-down measurements. Covalent post-translational modifications are localized regardless of their position in transmembrane domains. Top-down measurements provide a more detail oriented high-resolution description of post-transcriptional and post-translational diversity for enhanced understanding beyond genomic translation. PMID:21982782

Tunable integration of absorption-membrane-adsorption for efficiently separating low boiling gas mixtures near normal temperature

Science.gov (United States)

Liu, Huang; Pan, Yong; Liu, Bei; Sun, Changyu; Guo, Ping; Gao, Xueteng; Yang, Lanying; Ma, Qinglan; Chen, Guangjin

2016-01-01

Separation of low boiling gas mixtures is widely concerned in process industries. Now their separations heavily rely upon energy-intensive cryogenic processes. Here, we report a pseudo-absorption process for separating low boiling gas mixtures near normal temperature. In this process, absorption-membrane-adsorption is integrated by suspending suitable porous ZIF material in suitable solvent and forming selectively permeable liquid membrane around ZIF particles. Green solvents like water and glycol were used to form ZIF-8 slurry and tune the permeability of liquid membrane surrounding ZIF-8 particles. We found glycol molecules form tighter membrane while water molecules form looser membrane because of the hydrophobicity of ZIF-8. When using mixing solvents composed of glycol and water, the permeability of liquid membrane becomes tunable. It is shown that ZIF-8/water slurry always manifests remarkable higher separation selectivity than solid ZIF-8 and it could be tuned to further enhance the capture of light hydrocarbons by adding suitable quantity of glycol to water. Because of its lower viscosity and higher sorption/desorption rate, tunable ZIF-8/water-glycol slurry could be readily used as liquid absorbent to separate different kinds of low boiling gas mixtures by applying a multistage separation process in one traditional absorption tower, especially for the capture of light hydrocarbons. PMID:26892255

Reference: 673 [Arabidopsis Phenome Database[Archive

Lifescience Database Archive (English)

Full Text Available et al. 2007 Dec. J. Biol. Chem. 282(49):35945-53. Polar lipid trafficking is essential in eukaryotic cells as membranes... of lipid assembly are often distinct from final destination membranes. A striking example is the... biogenesis of the photosynthetic membranes (thylakoids) in plastids of plants. L...ipid biosynthetic enzymes at the endoplasmic reticulum and the inner and outer plastid envelope membranes ar

Functional update of the auxiliary proteins PsbW, PsbY, HCF136, PsbN, TerC and ALB3 in maintenance and assembly of PSII

Directory of Open Access Journals (Sweden)

Magdalena ePlöchinger

2016-04-01

Full Text Available Assembly of Photosystem (PS II in plants has turned out to be a highly complex process which, at least in part, occurs in a sequential order and requires many more auxiliary proteins than subunits present in the complex. Owing to the high evolutionary conservation of the subunit composition and the three-dimensional structure of the PSII complex, most plant factors involved in the biogenesis of PSII originated from cyanobacteria and only rarely evolved de novo. Furthermore, in chloroplasts the initial assembly steps occur in the non-appressed stroma lamellae, whereas the final assembly including the attachment of the major LHCII antenna proteins takes place in the grana regions. The stroma lamellae are also the place where part of PSII repair occurs, which very likely also involves assembly factors. In cyanobacteria initial PSII assembly also occurs in the thylakoid membrane, in so-called thylakoid centres, which are in contact with the plasma membrane. Here, we provide an update on the structures, localisations, topologies, functions, expression and interactions of the low molecular mass PSII subunits PsbY, PsbW and the auxiliary factors HCF136, PsbN, TerC and ALB3, assisting in PSII complex assembly and protein insertion into the thylakoid membrane.

Integrating seawater desalination and wastewater reclamation forward osmosis process using thin-film composite mixed matrix membrane with functionalized carbon nanotube blended polyethersulfone support layer.

Science.gov (United States)

Choi, Hyeon-Gyu; Son, Moon; Choi, Heechul

2017-10-01

Thin-film composite mixed matrix membrane (TFC MMM) with functionalized carbon nanotube (fCNT) blended in polyethersulfone (PES) support layer was synthesized via interfacial polymerization and phase inversion. This membrane was firstly tested in lab-scale integrating seawater desalination and wastewater reclamation forward osmosis (FO) process. Water flux of TFC MMM was increased by 72% compared to that of TFC membrane due to enhanced hydrophilicity. Although TFC MMM showed lower water flux than TFC commercial membrane, enhanced reverse salt flux selectivity (RSFS) of TFC MMM was observed compared to TFC membrane (15% higher) and TFC commercial membrane (4% higher), representing membrane permselectivity. Under effluent organic matter (EfOM) fouling test, 16% less normalized flux decline of TFC MMM was observed compared to TFC membrane. There was 8% less decline of TFC MMM compared to TFC commercial membrane due to fCNT effect on repulsive foulant-membrane interaction enhancement, caused by negatively charged membrane surface. After 10 min physical cleaning, TFC MMM displayed higher recovered normalized flux than TFC membrane (6%) and TFC commercial membrane (4%); this was also supported by visualized characterization of fouling layer. This study presents application of TFC MMM to integrated seawater desalination and wastewater reclamation FO process for the first time. It can be concluded that EfOM fouling of TFC MMM was suppressed due to repulsive foulant-membrane interaction. Copyright © 2017 Elsevier Ltd. All rights reserved.

A novel potassium channel in photosynthetic cyanobacteria.

Directory of Open Access Journals (Sweden)

Manuela Zanetti

Full Text Available Elucidation of the structure-function relationship of a small number of prokaryotic ion channels characterized so far greatly contributed to our knowledge on basic mechanisms of ion conduction. We identified a new potassium channel (SynK in the genome of the cyanobacterium Synechocystis sp. PCC6803, a photosynthetic model organism. SynK, when expressed in a K(+-uptake-system deficient E. coli strain, was able to recover growth of these organisms. The protein functions as a potassium selective ion channel when expressed in Chinese hamster ovary cells. The location of SynK in cyanobacteria in both thylakoid and plasmamembranes was revealed by immunogold electron microscopy and Western blotting of isolated membrane fractions. SynK seems to be conserved during evolution, giving rise to a TPK (two-pore K(+ channel family member which is shown here to be located in the thylakoid membrane of Arabidopsis. Our work characterizes a novel cyanobacterial potassium channel and indicates the molecular nature of the first higher plant thylakoid cation channel, opening the way to functional studies.

Endomembrane Cation Transporters and Membrane Trafficking

Energy Technology Data Exchange (ETDEWEB)

Sze, Heven [Univ. of Maryland, College Park, MD (United States). Dept. of Cell Biology & Molecular Genetics

2017-04-01

fertilization. Based on localization and mutant analyses, we conclude that CHXs modulate the ion balance, pH or both in micro-regions of endoplasmic reticulum, endosomes and prevacuolar compartment (PVC), and so influence membrane trafficking and signaling resulting in proper osmoregulation in guard cells and seed formation. We also demonstrated for the first time that AtKEA2 associates with chloroplasts, especially at the two poles of developing plastids. These results show that AtKEA1 and AtKEA2 transporters in specific microdomains of the inner envelope link local osmotic, ionic, and pH homeostasis to plastid division and thylakoid membrane formation. The first 3-D structure model of AtCHX was generated, and architecture-directed mutagenesis identified critical residues of the transport core giving insights to the transport mode of this family. Thus we have revealed for the first time crucial roles of an unknown K⁺/H⁺ transport family on plant growth (KEA), gas exchange, pollen cell wall, and different phases of reproduction (CHXs). The dynamic endomembrane of plant cells is integral to cytokinesis, cell expansion, defense, and cell wall formation, thus these studies are directly relevant to the mission of the Department of Energy and to a better understanding of determinants for enhancing plant biomass and plant tolerance to abiotic stress.

Atomic force microscopy studies of native photosynthetic membranes.

Science.gov (United States)

Sturgis, James N; Tucker, Jaimey D; Olsen, John D; Hunter, C Neil; Niederman, Robert A

2009-05-05

for quinones to diffuse freely. Measurement of the intercomplex distances between adjacent LH2 rings of Phaeospirillum molischianum has permitted the first calculation of the separation of bacteriochlorophyll a molecules in the native ICM. A recent AFM analysis of the organization of green plant photosystem II (PSII) in grana thylakoids revealed the protruding oxygen-evolving complex, crowded together in parallel alignment at three distinct levels of stacked membranes over the lumenal surface. The results also confirmed that PSII-LHCII supercomplexes are displaced relative to one another in opposing grana membranes.

An Integrated Framework Advancing Membrane Protein Modeling and Design.

Directory of Open Access Journals (Sweden)

Rebecca F Alford

2015-09-01

Full Text Available Membrane proteins are critical functional molecules in the human body, constituting more than 30% of open reading frames in the human genome. Unfortunately, a myriad of difficulties in overexpression and reconstitution into membrane mimetics severely limit our ability to determine their structures. Computational tools are therefore instrumental to membrane protein structure prediction, consequently increasing our understanding of membrane protein function and their role in disease. Here, we describe a general framework facilitating membrane protein modeling and design that combines the scientific principles for membrane protein modeling with the flexible software architecture of Rosetta3. This new framework, called RosettaMP, provides a general membrane representation that interfaces with scoring, conformational sampling, and mutation routines that can be easily combined to create new protocols. To demonstrate the capabilities of this implementation, we developed four proof-of-concept applications for (1 prediction of free energy changes upon mutation; (2 high-resolution structural refinement; (3 protein-protein docking; and (4 assembly of symmetric protein complexes, all in the membrane environment. Preliminary data show that these algorithms can produce meaningful scores and structures. The data also suggest needed improvements to both sampling routines and score functions. Importantly, the applications collectively demonstrate the potential of combining the flexible nature of RosettaMP with the power of Rosetta algorithms to facilitate membrane protein modeling and design.

Proteasome-mediated degradation of integral inner nuclear membrane protein emerin in fibroblasts lacking A-type lamins

International Nuclear Information System (INIS)

Muchir, Antoine; Massart, Catherine; Engelen, Baziel G. van; Lammens, Martin; Bonne, Gisele; Worman, Howard J.

2006-01-01

We previously identified and characterized a homozygous LMNA nonsense mutation leading to the absence of A-type lamins in a premature neonate who died at birth. We show here that the absence of A-type lamins is due to degradation of the aberrant mRNA transcript with a premature termination codon. In cultured fibroblasts from the subject with the homozygous LMNA nonsense mutation, there was a decreased steady-state expression of the integral inner nuclear membrane proteins emerin and nesprin-1α associated with their mislocalization to the bulk endoplasmic reticulum and a hyperphosphorylation of emerin. To determine if decreased emerin expression occurred post-translationally, we treated cells with a selective proteasome inhibitor and observed an increase in expression. Our results show that mislocalization of integral inner nuclear membrane proteins to the endoplasmic reticulum in human cells lacking A-type lamins leads to their degradation and provides the first evidence that their degradation is mediated by the proteasome

Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa.

Science.gov (United States)

Prapaiwan, N; Tharasanit, T; Punjachaipornpol, S; Yamtang, D; Roongsitthichai, A; Moonarmart, W; Kaeoket, K; Manee-In, S

2016-05-01

Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL) has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05) than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05). In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

Low-density Lipoprotein Improves Motility and Plasma Membrane Integrity of Cryopreserved Canine Epididymal Spermatozoa

Directory of Open Access Journals (Sweden)

N. Prapaiwan

2016-05-01

Full Text Available Cryopreservation of caudal epididymal spermatozoa is an effective technique to conserve genetic potentials of superior dogs when it is not possible to collect ejaculated spermatozoa. Although hen egg yolk is commonly supplemented into the semen extender, active substances within the egg yolk which protect sperm against cryoinjury remain to be discovered. Among its compositions, low-density lipoprotein (LDL has been reported to have a cryoprotective property for sperm cryopreservation. However, the effects of LDL on dog epididymal spermatozoa during cryopreservation have not yet been investigated. This study aimed to investigate the effects of LDL on epididymal spermatozoa quality following cryopreservation and thawing. After routine castration of 12 dogs, caudal epididymides from individuals were separated from the testes and cut into a few pieces in a Tris-buffer. Spermatozoa recovered from each sample were examined at once for sperm quality and divided into six groups of extender: no LDL, 20% egg yolk, 4%, 8%, 16%, and 24% LDL, before cryopreservation. The sperm aliquots were then equilibrated and conventionally frozen. After thawing, sperm motility, morphology, plasma membrane integrity, and acrosome integrity were evaluated. The results revealed that 4% LDL and 20% egg yolk yielded significantly higher sperm motility (57.69% and 52.69%, respectively, p<0.05 than other LDLs. In addition, 4% LDL yielded the significantly highest plasma membrane integrity (70.54%, p<0.05. In conclusion, the supplementation of 4% LDL in Tris-glucose extender could be applied for cryopreservation of canine epididymal spermatozoa.

Photoprotection vs. Photoinhibition of Photosystem II in Transplastomic Lettuce (Lactuca sativa) Dominantly Accumulating Astaxanthin.

Science.gov (United States)

Fujii, Ritsuko; Yamano, Nami; Hashimoto, Hideki; Misawa, Norihiko; Ifuku, Kentaro

2016-07-01

Transplastomic (chloroplast genome-modified; CGM) lettuce that dominantly accumulates astaxanthin grows similarly to a non-transgenic control with almost no accumulation of naturally occurring photosynthetic carotenoids. In this study, we evaluated the activity and assembly of PSII in CGM lettuce. The maximum quantum yield of PSII in CGM lettuce was <0.6; however, the quantum yield of PSII was comparable with that in control leaves under higher light intensity. CGM lettuce showed a lower ability to induce non-photochemical quenching (NPQ) than the control under various light intensities. The fraction of slowly recovering NPQ in CGM lettuce, which is considered to be photoinhibitory quenching (qI), was less than half that of the control. In fact, 1 O 2 generation was lower in CGM than in control leaves under high light intensity. CGM lettuce contained less PSII, accumulated mostly as a monomer in thylakoid membranes. The PSII monomers purified from the CGM thylakoids bound echinenone and canthaxanthin in addition to β-carotene, suggesting that a shortage of β-carotene and/or the binding of carbonyl carotenoids would interfere with the photophysical function as well as normal assembly of PSII. In contrast, high accumulation of astaxanthin and other carbonyl carotenoids was found within the thylakoid membranes. This finding would be associated with the suppression of photo-oxidative stress in the thylakoid membranes. Our observation suggests the importance of a specific balance between photoprotection and photoinhibition that can support normal photosynthesis in CGM lettuce producing astaxanthin. © The Author 2015. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Micro-coil NMR to monitor optimization of the reconstitution conditions for the integral membrane protein OmpW in detergent micelles

International Nuclear Information System (INIS)

Stanczak, Pawel; Zhang Qinghai; Horst, Reto; Serrano, Pedro; Wüthrich, Kurt

2012-01-01

Optimization of aqueous solutions of the integral membrane protein (IMP) OmpW for NMR structure determination has been monitored with micro-coil NMR, which enables the acquisition of NMR spectra using only micrograms of protein and detergent. The detergent 30-Fos (2-undecylphosphocholine) was found to yield the best 2D [ 15 N, 1 H]-TROSY correlation NMR spectra of [ 2 H, 15 N]-labeled OmpW. For the OmpW structure determination we then optimized the 30-Fos concentration, the sample temperature and long-time stability, and the deuteration level of the protein. Some emerging guidelines for reconstitution of β-barrel integral membrane proteins in structural biology are discussed.

3D pressure field in lipid membranes and membrane-protein complexes

DEFF Research Database (Denmark)

Ollila, O H Samuli; Risselada, H Jelger; Louhivuori, Martti

2009-01-01

We calculate full 3D pressure fields for inhomogeneous nanoscale systems using molecular dynamics simulation data. The fields represent systems with increasing level of complexity, ranging from semivesicles and vesicles to membranes characterized by coexistence of two phases, including also...... a protein-membrane complex. We show that the 3D pressure field is distinctly different for curved and planar bilayers, the pressure field depends strongly on the phase of the membrane, and that an integral protein modulates the tension and elastic properties of the membrane....

Diffusion of Integral Membrane Proteins in Protein-Rich Membranes

Czech Academy of Sciences Publication Activity Database

Javanainen, M.; Martinez-Seara, Hector; Metzler, R.; Vattulainen, I.

2017-01-01

Roč. 8, č. 17 (2017), s. 4308-4313 ISSN 1948-7185 R&D Projects: GA ČR(CZ) GBP208/12/G016 Institutional support: RVO:61388963 Keywords : giant unilamellar vesicles * single-molecule tracking * lipid bilayer membranes Subject RIV: CF - Physical ; Theoretical Chemistry OBOR OECD: Physical chemistry Impact factor: 9.353, year: 2016

Is chloroplast import of photosynthesis proteins facilitated by an actin-TOC-TIC-VIPP1 complex?

Science.gov (United States)

Jouhet, Juliette; Gray, John C

2009-10-01

Actin filaments are major components of the cytoskeleton that interact with chloroplast envelope membranes to allow chloroplast positioning and movement, stromule mobility and gravitropism perception. We recently reported that Toc159, a component of the TOC complex of the chloroplast protein import apparatus, interacts directly with actin. The interaction of Toc159 and actin was identified by co-immunoprecipitation and co-sedimentation experiments with detergent-solubilised pea chloroplast envelope membranes. In addition, many of the components of the TOC-TIC protein import apparatus and VIPP1 (vesicle-inducing protein in plastids 1) were identified by mass spectroscopy in the material co-immunoprecipitated with antibodies to actin. Toc159 is the receptor for the import of photosynthesis proteins and VIPP1 is involved in thylakoid membrane formation by inducing vesicle formation from the chloroplast inner envelope membrane, suggesting we may have identified an actin-TOC-TIC-VIPP1 complex that may provide a means of channeling cytosolic preproteins to the thylakoid membrane. The interaction of Toc159 with actin may facilitate exchange between the putative soluble and membrane forms of Toc159 and promote the interaction of cytosolic preproteins with the TOC complex.

Membrane Bioreactor (MBR) Technology for Wastewater Treatment and Reclamation: Membrane Fouling.

Science.gov (United States)

Iorhemen, Oliver Terna; Hamza, Rania Ahmed; Tay, Joo Hwa

2016-06-15

The membrane bioreactor (MBR) has emerged as an efficient compact technology for municipal and industrial wastewater treatment. The major drawback impeding wider application of MBRs is membrane fouling, which significantly reduces membrane performance and lifespan, resulting in a significant increase in maintenance and operating costs. Finding sustainable membrane fouling mitigation strategies in MBRs has been one of the main concerns over the last two decades. This paper provides an overview of membrane fouling and studies conducted to identify mitigating strategies for fouling in MBRs. Classes of foulants, including biofoulants, organic foulants and inorganic foulants, as well as factors influencing membrane fouling are outlined. Recent research attempts on fouling control, including addition of coagulants and adsorbents, combination of aerobic granulation with MBRs, introduction of granular materials with air scouring in the MBR tank, and quorum quenching are presented. The addition of coagulants and adsorbents shows a significant membrane fouling reduction, but further research is needed to establish optimum dosages of the various coagulants/adsorbents. Similarly, the integration of aerobic granulation with MBRs, which targets biofoulants and organic foulants, shows outstanding filtration performance and a significant reduction in fouling rate, as well as excellent nutrients removal. However, further research is needed on the enhancement of long-term granule integrity. Quorum quenching also offers a strong potential for fouling control, but pilot-scale testing is required to explore the feasibility of full-scale application.

A microbial fuel cell–membrane bioreactor integrated system for cost-effective wastewater treatment

International Nuclear Information System (INIS)

Wang, Yong-Peng; Liu, Xian-Wei; Li, Wen-Wei; Li, Feng; Wang, Yun-Kun; Sheng, Guo-Ping; Zeng, Raymond J.; Yu, Han-Qing

2012-01-01

Highlights: ► An MFC–MBR integrated system for wastewater treatment and electricity generation. ► Stable electricity generation during 1000-h continuous operation. ► Low-cost electrode, separator and filter materials were adopted. -- Abstract: Microbial fuel cell (MFC) and membrane bioreactor (MBR) are both promising technologies for wastewater treatment, but both with limitations. In this study, a novel MFC–MBR integrated system, which combines the advantages of the individual systems, was proposed for simultaneous wastewater treatment and energy recovery. The system favored a better utilization of the oxygen in the aeration tank of MBR by the MFC biocathode, and enabled a high effluent quality. Continuous and stable electricity generation, with the average current of 1.9 ± 0.4 mA, was achieved over a long period of about 40 days. The maximum power density reached 6.0 W m −3 . Moreover, low-cost materials were used for the reactor construction. This integrated system shows great promise for practical wastewater treatment application.

An integrated field-effect microdevice for monitoring membrane transport in Xenopus laevis oocytes via lateral proton diffusion.

Directory of Open Access Journals (Sweden)

Daniel Felix Schaffhauser

Full Text Available An integrated microdevice for measuring proton-dependent membrane activity at the surface of Xenopus laevis oocytes is presented. By establishing a stable contact between the oocyte vitelline membrane and an ion-sensitive field-effect (ISFET sensor inside a microperfusion channel, changes in surface pH that are hypothesized to result from facilitated proton lateral diffusion along the membrane were detected. The solute diffusion barrier created between the sensor and the active membrane area allowed detection of surface proton concentration free from interference of solutes in bulk solution. The proposed sensor mechanism was verified by heterologously expressing membrane transport proteins and recording changes in surface pH during application of the specific substrates. Experiments conducted on two families of phosphate-sodium cotransporters (SLC20 & SLC34 demonstrated that it is possible to detect phosphate transport for both electrogenic and electroneutral isoforms and distinguish between transport of different phosphate species. Furthermore, the transport activity of the proton/amino acid cotransporter PAT1 assayed using conventional whole cell electrophysiology correlated well with changes in surface pH, confirming the ability of the system to detect activity proportional to expression level.

Arabidopsis Tic40 expression in tobacco chloroplasts results in massive proliferation of the inner envelope membrane and upregulation of associated proteins.

Science.gov (United States)

Singh, Nameirakpam Dolendro; Li, Ming; Lee, Sueng-Bum; Schnell, Danny; Daniell, Henry

2008-12-01

The chloroplast inner envelope membrane (IM) plays essential roles in lipid synthesis, metabolite transport, and cellular signaling in plants. We have targeted a model nucleus-encoded IM protein from Arabidopsis thaliana, pre-Tic40-His, by relocating its expression from the nucleus to the chloroplast genome. Pre-Tic40-His was properly targeted, processed, and inserted. It attained correct topology and was folded and assembled into a TIC complex, where it accounted for up to 15% of the total chloroplast protein. These results confirm the existence of a novel pathway for protein targeting to the IM. Tic40-His overexpression resulted in a massive proliferation of the IM (up to 19 layers in electron micrographs) without significant effects on plant growth or reproduction. Consistent with IM proliferation, the expression levels of other endogenous IM proteins (IEP37, PPT, Tic110) were significantly (10-fold) upregulated but those of outer envelope membrane (Toc159), stromal (hsp93, cpn60), or thylakoid (LHCP, OE23) proteins were not increased, suggesting retrograde signal transduction between chloroplast and nuclear genomes to increase lipid and protein components for accommodation of increased accumulation of Tic40. This study opens the door for understanding the regulation of membrane biogenesis within the organelle and the utilization of transgenic chloroplasts as bioreactors for hyperaccumulation of membrane proteins for biotechnological applications.

[Effect of damage integrity rat brain synaptic membranes on the functional activity GABA(A)-receptor/Cl(-)-ionophore complex in the CNC].

Science.gov (United States)

Rebrov, I G; Kalinina, M V

2013-01-01

Functional activity of the CGABA(A)-receptor/Cl(-) ionophore complex was investigated the muscimol-stimulated entry of the radioactive isotope 36Cl(-) in synaptoneurosomes in changing the structure and permeability of neuronal membranes. Integrity of the membranes was damaged by removal of Ca(+2) and Mg(+2) from the incubation medium and by the method of freezing-thawing synaptoneurosomes. In both cases, an increase in basal 36Cl(-) entry into synaptoneurosomes, indicating increased nonspecific permeability of neuronal membranes, and decreased activity the CABA(A)-receptor/Cl(-) ionophore complex. The conclusion about the relationship of processes damage neuronal membranes and reducing the inhibitory processes in the epileptic focus.

Multichannel detection of ionic currents through two nanopores fabricated on integrated Si3N4 membranes.

Science.gov (United States)

Yanagi, Itaru; Akahori, Rena; Aoki, Mayu; Harada, Kunio; Takeda, Ken-Ichi

2016-08-16

Integration of solid-state nanopores and multichannel detection of signals from each nanopore are effective measures for realizing high-throughput nanopore sensors. In the present study, we demonstrated fabrication of Si3N4 membrane arrays and the simultaneous measurement of ionic currents through two nanopores formed in two adjacent membranes. Membranes with thicknesses as low as 6.4 nm and small nanopores with diameters of less than 2 nm could be fabricated using the poly-Si sacrificial-layer process and multilevel pulse-voltage injection. Using the fabricated nanopore membranes, we successfully achieved simultaneous detection of clear ionic-current blockades when single-stranded short homopolymers (poly(dA)60) passed through two nanopores. In addition, we investigated the signal crosstalk and leakage current among separated chambers. When two nanopores were isolated on the front surface of the membrane, there was no signal crosstalk or leakage current between the chambers. However, when two nanopores were isolated on the backside of the Si substrate, signal crosstalk and leakage current were observed owing to high-capacitance coupling between the chambers and electrolysis of water on the surface of the Si substrate. The signal crosstalk and leakage current could be suppressed by oxidizing the exposed Si surface in the membrane chip. Finally, the observed ionic-current blockade when poly(dA)60 passed through the nanopore in the oxidized chip was approximately half of that observed in the non-oxidized chip.

Hydrogen Selective Exfoliated Zeolite Membranes

Energy Technology Data Exchange (ETDEWEB)

Tsapatsis, Michael [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Daoutidis, Prodromos [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Elyassi, Bahman [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Lima, Fernando [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Iyer, Aparna [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Agrawal, Kumar [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science; Sabnis, Sanket [Univ. of Minnesota, Minneapolis, MN (United States). Department of Chemical Engineering and Materials Science

2015-04-06

The objective of this project was to develop and evaluate an innovative membrane technology at process conditions that would be representative of Integrated Gasification Combined Cycle (IGCC) advanced power generation with pre-combustion capture of carbon dioxide (CO₂). This research focused on hydrogen (H₂)-selective zeolite membranes that could be utilized to separate conditioned syngas into H₂-rich and CO₂-rich components. Both experiments and process design and optimization calculations were performed to evaluate the concept of ultra-thin membranes made from zeolites nanosheets. In this work, efforts in the laboratory were made to tackle two fundamental challenges in application of zeolite membranes in harsh industrial environments, namely, membrane thickness and membrane stability. Conventional zeolite membranes have thicknesses in the micron range, limiting their performance. In this research, we developed a method for fabrication of ultimately thin zeolite membranes based on zeolite nanosheets. A range of layered zeolites (MWW, RWR, NSI structure types) suitable for hydrogen separation was successfully exfoliated to their constituent nanosheets. Further, membranes were made from one of these zeolites, MWW, to demonstrate the potential of this group of materials. Moreover, long-term steam stability of these zeolites (up to 6 months) was investigated in high concentrations of steam (35 mol% and 95 mole%), high pressure (10 barg), and high temperatures (350 °C and 600 °C) relevant to conditions of water-gas-shift and steam methane reforming reactions. It was found that certain nanosheets are stable, and that stability depends on the concentration of structural defects. Additionally, models that represent a water-gas-shift (WGS) membrane reactor equipped with the zeolite membrane were developed for systems studies. These studies had the aim of analyzing the effect of the membrane reactor integration into IGCC plants

Novel process integration for biodiesel blend in membrane reactive divided wall (MRDW column

Directory of Open Access Journals (Sweden)

Sakhre Vandana

2016-03-01

Full Text Available The paper proposes a novel process integration for biodiesel blend in the Membrane assisted Reactive Divided Wall Distillation (MRDW column. Biodiesel is a green fuel and grade of biodiesel blend is B20 (% which consist of 20% biodiesel and rest 80% commercial diesel. Instead of commercial diesel, Tertiary Amyl Ethyl Ether (TAEE was used as an environment friendly fuel for blending biodiesel. Biodiesel and TAEE were synthesized in a pilot scale reactive distillation column. Dual reactive distillation and MRDW were simulated using aspen plus. B20 (% limit calculation was performed using feed flow rates of both TAEE and biodiesel. MRDW was compared with dual reactive distillation column and it was observed that MRDW is comparatively cost effective and suitable in terms of improved heat integration and flow pattern.

VIABILITY AND PLASMA MEMBRANE INTEGRITY OF THE SPOTTED BUFFALO EPIDIDYMAL SPERMATOZOA AFTER THAWING WITH THE ADDITION OF DEXTROSE INTO THE EXTENDER

Directory of Open Access Journals (Sweden)

M. RIZAL

2009-01-01

Full Text Available h e objective of this study was to obtain the viability and plasma membrane integrity of the spotted buff alo epididymal sperm after addition of dextrose into Andromed® extender. Spermatozoa that have been collected from cauda epididymis were diluted with Andromed® extender as control (K and Andromed® + 0.2% dextrose (P1 and Andromed® + 0.4% dextrose (P2 as treatments. h e results showed that the quality of epididymal spermatozoa decreased during cryopreservation process. h e percentage of motility after thawing in P1 (46% and P2 (46.67% were signifi cantly higher (P<0.05 compared to K (41% as well as the percentage of live sperm in P1 (58.8% and P2 (60% compared to K (52.2%. h e percentage of membrane integrity in P1, P2 and K were 67.4; 66.8 and 68 %, respectively. In conclusion, the addition of 0.2 and 0.4% of dextrose into Andromed® acted as an extra cellular cryoprotectant and could maintain the viability and membrane integrity of the spotted buff alo epididymal spermatozoa after thawing.

Integrated distillation-membrane process for bio-ethanol and bio-butanol recovery from actual fermentation broths: Separation energy efficiency and fate of secondary fermentation products

Science.gov (United States)

A hybrid process integrating vapor stripping with vapor compression and vapor permeation membrane separation, termed Membrane Assisted Vapor Stripping (MAVS), was evaluated for recovery and dehydration of ethanol and/or 1-butanol from aqueous solution as an alternative to convent...

A membrane stirrer for product recovery and substrate feeding.

Science.gov (United States)

Femmer, T; Carstensen, F; Wessling, M

2015-02-01

During fermentation processes, in situ product recovery (ISPR) using submerged membranes allows a continuous operation mode with effective product removal. Continuous recovery reduces product inhibition and organisms in the reactor are not exposed to changing reaction conditions. For an effective in situ product removal, submerged membrane systems should have a sufficient large membrane area and an anti-fouling concept integrated in a compact device for the limited space in a lab-scale bioreactor. We present a new membrane stirrer with integrated filtration membranes on the impeller blades as well as an integrated gassing concept in an all-in-one device. The stirrer is fabricated by rapid prototyping and is equipped with a commercial micromesh membrane. Filtration performance is tested using a yeast cell suspension with different stirring speeds and aeration fluxes. We reduce membrane fouling by backflushing through the membrane with the product stream. © 2014 Wiley Periodicals, Inc.

Membrane Bioreactor (MBR Technology for Wastewater Treatment and Reclamation: Membrane Fouling

Directory of Open Access Journals (Sweden)

Oliver Terna Iorhemen

2016-06-01

Full Text Available The membrane bioreactor (MBR has emerged as an efficient compact technology for municipal and industrial wastewater treatment. The major drawback impeding wider application of MBRs is membrane fouling, which significantly reduces membrane performance and lifespan, resulting in a significant increase in maintenance and operating costs. Finding sustainable membrane fouling mitigation strategies in MBRs has been one of the main concerns over the last two decades. This paper provides an overview of membrane fouling and studies conducted to identify mitigating strategies for fouling in MBRs. Classes of foulants, including biofoulants, organic foulants and inorganic foulants, as well as factors influencing membrane fouling are outlined. Recent research attempts on fouling control, including addition of coagulants and adsorbents, combination of aerobic granulation with MBRs, introduction of granular materials with air scouring in the MBR tank, and quorum quenching are presented. The addition of coagulants and adsorbents shows a significant membrane fouling reduction, but further research is needed to establish optimum dosages of the various coagulants/adsorbents. Similarly, the integration of aerobic granulation with MBRs, which targets biofoulants and organic foulants, shows outstanding filtration performance and a significant reduction in fouling rate, as well as excellent nutrients removal. However, further research is needed on the enhancement of long-term granule integrity. Quorum quenching also offers a strong potential for fouling control, but pilot-scale testing is required to explore the feasibility of full-scale application.

Integral design method for simple and small Mars lander system using membrane aeroshell

Science.gov (United States)

Sakagami, Ryo; Takahashi, Ryohei; Wachi, Akifumi; Koshiro, Yuki; Maezawa, Hiroyuki; Kasai, Yasko; Nakasuka, Shinichi

2018-03-01

To execute Mars surface exploration missions, spacecraft need to overcome the difficulties of the Mars entry, descent, and landing (EDL) sequences. Previous landing missions overcame these challenges with complicated systems that could only be executed by organizations with mature technology and abundant financial resources. In this paper, we propose a novel integral design methodology for a small, simple Mars lander that is achievable even by organizations with limited technology and resources such as universities or emerging countries. We aim to design a lander (including its interplanetary cruise stage) whose size and mass are under 1 m3 and 150 kg, respectively. We adopted only two components for Mars EDL process: a "membrane aeroshell" for the Mars atmospheric entry and descent sequence and one additional mechanism for the landing sequence. The landing mechanism was selected from the following three candidates: (1) solid thrusters, (2) aluminum foam, and (3) a vented airbag. We present a reasonable design process, visualize dependencies among parameters, summarize sizing methods for each component, and propose the way to integrate these components into one system. To demonstrate the effectiveness, we applied this methodology to the actual Mars EDL mission led by the National Institute of Information and Communications Technology (NICT) and the University of Tokyo. As a result, an 80 kg class Mars lander with a 1.75 m radius membrane aeroshell and a vented airbag was designed, and the maximum landing shock that the lander will receive was 115 G.

Effect of acclimation medium on cell viability, membrane integrity and ability to consume malic acid in synthetic wine by oenological Lactobacillus plantarum strains.

Science.gov (United States)

Bravo-Ferrada, B M; Tymczyszyn, E E; Gómez-Zavaglia, A; Semorile, L

2014-02-01

The aim of this work was to evaluate the effect of acclimation on the viability, membrane integrity and the ability to consume malic acid of three oenological strains of Lactobacillus plantarum. Cultures in the stationary phase were inoculated in an acclimation medium (Accl.) containing 0, 6 or 10% v/v ethanol and incubated 48 h at 28°C. After incubation, cells were harvested by centrifugation and inoculated in a synthetic wine, containing 14% v/v ethanol and pH 3.5 at 28°C. Viability and membrane integrity were determined by flow cytometry (FC) using carboxyfluorescein diacetate (cFDA) and propidium iodide. Bacterial growth and malic acid consumption were monitored in a synthetic wine during 15 days. In nonacclimated strains, the damage of bacterial membranes produced a dramatic decrease in microbial viability in synthetic wine. In contrast, survival of strains previously acclimated in Accl. with 6 and 10% v/v ethanol was noticeable higher. Therefore, acclimation with ethanol increased the cultivability in synthetic wine and consequently, the consumption of l-malic acid after 15 days of growth. Acclimation of oenological strains in media containing ethanol prior to wine inoculation significantly decreases the membrane damage and improves viability in the harsh wine conditions. The role of membrane integrity is crucial to warrant the degradation of l-malic acid. The efficiency of multiparametric FC in monitoring viability and membrane damage along with the malic acid consumption has a strong impact on winemaking because it represents a useful tool for a quick and highly reliable evaluation of oenological parameters. © 2013 The Society for Applied Microbiology.

Comparison and analysis of membrane fouling between flocculent sludge membrane bioreactor and granular sludge membrane bioreactor.

Directory of Open Access Journals (Sweden)

Wang Jing-Feng

Full Text Available The goal of this study is to investigate the effect of inoculating granules on reducing membrane fouling. In order to evaluate the differences in performance between flocculent sludge and aerobic granular sludge in membrane reactors (MBRs, two reactors were run in parallel and various parameters related to membrane fouling were measured. The results indicated that specific resistance to the fouling layer was five times greater than that of mixed liquor sludge in the granular MBR. The floc sludge more easily formed a compact layer on the membrane surface, and increased membrane resistance. Specifically, the floc sludge had a higher moisture content, extracellular polymeric substances concentration, and negative surface charge. In contrast, aerobic granules could improve structural integrity and strength, which contributed to the preferable permeate performance. Therefore, inoculating aerobic granules in a MBR presents an effective method of reducing the membrane fouling associated with floc sludge the perspective of from the morphological characteristics of microbial aggregates.

Improving lactic acid productivity from wheat straw hydrolysates by membrane integrated repeated batch fermentation under non-sterilized conditions

DEFF Research Database (Denmark)

Zhang, Yuming; Chen, Xiangrong; Qi, Benkun

2014-01-01

to eliminate the sequential utilization of mixed sugar and feedback inhibition during batch fermentation, membrane integrated repeated batch fermentation (MIRB) was used to improve LA productivity. With MIRB, a high cell density was obtained and the simultaneous fermentation of glucose, xylose and arabinose...

Role of ions in the regulation of light-harvesting

Directory of Open Access Journals (Sweden)

Radek Kana

2016-12-01

Full Text Available Regulation of photosynthetic light harvesting in the thylakoids is one of the major key factors affecting the efficiency of photosynthesis. Thylakoid membrane is negatively charged and influences both the structure and the function of the primarily photosynthetic reactions through its electrical double layer. Further, there is a heterogeneous organization of soluble ions (K+, Mg2+, Cl- attached to the thylakoid membrane that, together with fixed charges (negatively charged amino acids, lipids, provides an electrical field. The electrical double layer is affected by the valence of the ions and interferes with the regulation of state transitions, protein interactions, and excitation energy spillover from Photosystem II to Photosystem I. These effects are reflected in changes in the intensity of chlorophyll a fluorescence, which is also a measure of photoprotective non-photochemical quenching of the excited state of chlorophyll a. A triggering of non-photochemical quenching proceeds via lumen acidification and is coupled to the export of positive counter-ions (Mg2+, K+ to the stroma or/and negative ions (e.g., Cl- into the lumen. The effect of protons and anions in the lumen and of the cations (Mg2+, K+ in the stroma are, thus, functionally tightly interconnected. In this review, we discuss the consequences of the model of electrical double layer, proposed by James Barber (J. Barber (1980 Biochim Biophys Acta 594:253-308 in light of light-harvesting regulation. Further, we explain differences between electrostatic screening and neutralization, and we emphasize the opposite effect of monovalent (K+ and divalent (Mg2+ ions on light-harvesting and on screening of the negative charges on the thylakoid membrane; this effect needs to be incorporated in all future models of photosynthetic regulation by ion channels and transporters.

Influence of estrogenic pesticides on membrane integrity and membrane transfer of monosaccharide into the human red cell

International Nuclear Information System (INIS)

Ingermann, R.L.

1989-01-01

Some natural and synthetic estrogens inhibit carrier-mediated transport of glucose into human red blood cells and membrane vesicles from the placenta. The inhibitory action of these estrogens on transport appears to be a direct effect at the membrane and does not involve receptor binding and protein synthesis. It is not clear, however, whether such inhibition is a common feature among estrogenic agents. Several chlorinated hydrocarbon pesticides have been shown to possess estrogenic activity. These pesticides could have inhibitory effects on the human sodium-independent glucose transporter. Owing to the apparent importance of this membrane transporter in human tissues, direct interaction of hormones and xenobiotics with the glucose transporter is of fundamental significance. Some pesticides have been shown to alter membrane structure directly and alter the passive permeability of membranes. Whether the estrogenic pesticides influence passive diffusion of sugars across membranes has not been established. Finally, preliminary observations have suggested that some estrogens and pesticides have lytic effects on intact cells. Consequently, this study focuses on the ability of several estrogens and estrogenic pesticides to disrupt the cell membrane, influence the monosaccharide transporter, and alter the rate of monosaccharide permeation through the membrane by simple diffusion

A Proteomics Approach to Membrane Trafficking

NARCIS (Netherlands)

Groen, A.J.; Vries, de S.C.; Lilley, K.S.

2008-01-01

Membrane trafficking, including that of integral membrane proteins as well as peripherally associated proteins, appears to be a vital process common to all eukaryotes. An important element of membrane trafficking is to determine the protein composition of the various endomembrane compartments. A

Efficient production of membrane-integrated and detergent-soluble G protein-coupled receptors in Escherichia coli.

Science.gov (United States)

Link, A James; Skretas, Georgios; Strauch, Eva-Maria; Chari, Nandini S; Georgiou, George

2008-10-01

G protein-coupled receptors (GPCRs) are notoriously difficult to express, particularly in microbial systems. Using GPCR fusions with the green fluorescent protein (GFP), we conducted studies to identify bacterial host effector genes that result in a general and significant enhancement in the amount of membrane-integrated human GPCRs that can be produced in Escherichia coli. We show that coexpression of the membrane-bound AAA+ protease FtsH greatly enhances the expression yield of four different class I GPCRs, irrespective of the presence of GFP. Using this new expression system, we produced 0.5 and 2 mg/L of detergent-solubilized and purified full-length central cannabinoid receptor (CB1) and bradykinin receptor 2 (BR2) in shake flask cultures, respectively, two proteins that had previously eluded expression in microbial systems.

MUNI Ways and Structures Building Integrated Solar Membrane Project

Energy Technology Data Exchange (ETDEWEB)

Smith, Randall

2014-07-03

The initial goal of the MUNI Ways and Structures Building Integrated Solar Membrane Installation Project was for the City and County of San Francisco (CCSF) to gain experience using the integrated higher efficiency solar photovoltaic (PV) single-ply membrane product, as it differs from the conventional, low efficiency, thin-film PV products, to determine the feasibility of success of larger deployment. As several of CCSF’s municipal rooftops are constrained with respect to weight restrictions, staff of the Energy Generation Group of the San Francisco Public Utilities Commission (SFPUC) proposed to install a solar PV system using single-ply membrane The installation of the 100 kW (DC-STC) lightweight photo voltaic (PV) system at the MUNI Ways and Structures Center (700 Pennsylvania Ave., San Francisco) is a continuation of the commitment of the City and County of San Francisco (CCSF) to increase the pace of municipal solar development, and serve its municipal facilities with clean renewable energy. The fourteen (14) solar photovoltaic systems that have already been installed at CCSF municipal facilities are assisting in the reduction of fossil-fuel use, and reduction of greenhouse gases from fossil combustion. The MUNI Ways & Structures Center roof has a relatively low weight-bearing capacity (3.25 pounds per square foot) and use of traditional crystalline panels was therefore rejected. Consequently it was decided to use the best available highest efficiency Building-Integrated PV (BIPV) technology, with consideration for reliability and experience of the manufacturer which can meet the low weight-bearing capacity criteria. The original goal of the project was to provide an opportunity to monitor the results of the BIPV technology and compare these results to other City and County of San Francisco installed PV systems. The MUNI Ways and Structures Center was acquired from the Cookson Doors Company, which had run the Center for many decades. The building was

Spacesuit Water Membrane Evaporator Integration with the ISS Extravehicular Mobility Unit

Science.gov (United States)

Margiott, Victoria; Boyle, Robert

2014-01-01

NASA has developed a Solid Water Membrane Evaporation (SWME) to provide cooling for the next generation spacesuit. The current spacesuit team has looked at this technology from the standpoint of using the ISS EMU to demonstrate the SWME technology while EVA, and from the standpoint of augmenting EMU cooling in the case of a fouled EMU cooling system. One approach to increasing the TRL of the system is to incorporate this hardware with the existing EMU. Several integration issues were addressed to support a potential demonstration of the SWME with the existing EMU. Systems analysis was performed to assess the capability of the SWME to maintain crewmember cooling and comfort as a replacement for sublimation. The materials of the SWME were reviewed to address compatibility with the EMU. Conceptual system placement and integration with the EMU via an EVA umbilical system to ensure crew mobility and Airlock egress were performed. A concept of operation for EVA use was identified that is compatible with the existing system. This concept is extensible as a means to provide cooling for the existing EMU. The cooling system of one of the EMUs on orbit has degraded, with the root cause undetermined. Should there be a common cause resident on ISS, this integration could provide a means to recover cooling capability for EMUs on orbit.

Annexin A4 and A6 induce membrane curvature and constriction during cell membrane repair

DEFF Research Database (Denmark)

Boye, Theresa Louise; Maeda, Kenji; Pezeshkian, Weria

2017-01-01

Efficient cell membrane repair mechanisms are essential for maintaining membrane integrity and thus for cell life. Here we show that the Ca2+- and phospholipid-binding proteins annexin A4 and A6 are involved in plasma membrane repair and needed for rapid closure of micron-size holes. We demonstrate...... that annexin A4 binds to artificial membranes and generates curvature force initiated from free edges, whereas annexin A6 induces constriction force. In cells, plasma membrane injury and Ca2+ influx recruit annexin A4 to the vicinity of membrane wound edges where its homo-trimerization leads to membrane...... that induction of curvature force around wound edges is an early key event in cell membrane repair....

Isolation of Highly Purified Fractions of Plasma Membrane and Tonoplast from the Same Homogenate of Soybean Hypocotyls by Free-Flow Electrophoresis 1

Science.gov (United States)

Sandelius, Anna Stina; Penel, Claude; Auderset, Guy; Brightman, Andrew; Millard, Merle; Morré, D. James

1986-01-01

A procedure is described whereby highly purified fractions of plasma membrane and tonoplast were isolated from hypocotyls of dark-grown soybean (Glycine max L. var Wayne) by the technique of preparative free-flow electrophoresis. Fractions migrating the slowest toward the anode were enriched in thick (10 nanometers) membranes identified as plasma membranes based on ability to bind N-1-naphthylphthalamic acid (NPA), glucan synthetase-II, and K+-stimulated, vanadate-inhibited Mg2+ ATPase, reaction with phosphotungstic acid at low pH on electron microscope sections, and morphological evaluations. Fractions migrating farthest toward the anode (farthest from the point of sample injection) were enriched in membrane vesicles with thick (7-9 nanometers) membranes that did not stain with phosphotungstic acid at low pH, contained a nitrate-inhibited, Cl-stimulated ATPase and had the in situ morphological characteristics of tonoplast including the presence of flocculent contents. These vesicles neither bound NPA nor contained levels of glucan synthetase II above background. Other membranous cell components such as dictyosomes (fucosyltransferase, latent nucleosidediphosphate phosphatase), endoplasmic reticulum vesicles (NADH- and NADPH- cytochrome c reductase), mitochondria (succinate-2(p-indophenyl)-3-p-nitrophenyl)-5-phenyl tetrazolium-reductase and cytochrome oxidase) and plastids (carotenoids and monogalactosyl diglyceride synthetase) were identified on the basis of appropriate marker constituents and, except for plastid thylakoids, had thin (marker activities. From electron microscope morphometry (using both membrane measurements and staining with phosphotungstic acid at low pH) and analysis of marker enzymes, both plasma membrane and tonoplast fractions were estimated to be about 90% pure. Neither fraction appeared to be contaminated by the other by more than 3%. Images Fig. 1 Fig. 3 Fig. 4 Fig. 5 Fig. 9 PMID:16664771

Relative Abundance of Integral Plasma Membrane Proteins in Arabidopsis Leaf and Root Tissue Determined by Metabolic Labeling and Mass Spectrometry

Science.gov (United States)

Bernfur, Katja; Larsson, Olaf; Larsson, Christer; Gustavsson, Niklas

2013-01-01

Metabolic labeling of proteins with a stable isotope (15N) in intact Arabidopsis plants was used for accurate determination by mass spectrometry of differences in protein abundance between plasma membranes isolated from leaves and roots. In total, 703 proteins were identified, of which 188 were predicted to be integral membrane proteins. Major classes were transporters, receptors, proteins involved in membrane trafficking and cell wall-related proteins. Forty-one of the integral proteins, including nine of the 13 isoforms of the PIP (plasma membrane intrinsic protein) aquaporin subfamily, could be identified by peptides unique to these proteins, which made it possible to determine their relative abundance in leaf and root tissue. In addition, peptides shared between isoforms gave information on the proportions of these isoforms. A comparison between our data for protein levels and corresponding data for mRNA levels in the widely used database Genevestigator showed an agreement for only about two thirds of the proteins. By contrast, localization data available in the literature for 21 of the 41 proteins show a much better agreement with our data, in particular data based on immunostaining of proteins and GUS-staining of promoter activity. Thus, although mRNA levels may provide a useful approximation for protein levels, detection and quantification of isoform-specific peptides by proteomics should generate the most reliable data for the proteome. PMID:23990937

Removal of Pharmaceutical and Personal Care Products (PPCPs) from Municipal Waste Water with Integrated Membrane Systems, MBR-RO/NF.

Science.gov (United States)

Wang, Yonggang; Wang, Xu; Li, Mingwei; Dong, Jing; Sun, Changhong; Chen, Guanyi

2018-02-05

This study focuses on the application of combining membrane bioreactor (MBR) treatment with reverse osmosis (RO) or nanofiltration (NF) membrane treatment for removal of pharmaceuticals and personal care products (PPCPs) in municipal wastewater. Twenty-seven PPCPs were measured in real influent with lowest average concentration being trimethoprim (7.12 ng/L) and the highest being caffeine (18.4 ng/L). The results suggest that the MBR system effectively removes the PPCPs with an efficiency of between 41.08% and 95.41%, and that the integrated membrane systems, MBR-RO/NF, can achieve even higher removal rates of above 95% for most of them. The results also suggest that, due to the differences in removal mechanisms of NF/RO membrane, differences of removal rates exist. In this study, the combination of MBR-NF resulted in the removal of 13 compounds to below detection limits and MBR-RO achieved even better results with removal of 20 compounds to below detection limits.

Studies of radiation induced membrane damage in lymphocytes using fluorescent probes

International Nuclear Information System (INIS)

Nikesch, W.

1974-01-01

The fluorescent probes perylene (PER), 1-anilino-8-naphthalene sulfonic acid (ANS), and fluorescein diacetate (FDA) were used to investigate membrane changes caused by ionizing radiation. Probe response to various other perturbations (variation of pH, temperature, and salt concentration, and treatment with phythohemagglutinin (PHA) and saponins) was also investigated to better understand membrane-probe interactions. ANS was used to probe the membrane surface, PER to probe the membrane interior, and FDA to investigate membrane integrity. Polarization of fluorescent light from ANS and PER was used to investigate the microviscosity and order of the membrane surface and interior respectively. Irradiated cells (600 R) were shown to have a decreased rate of hydrolysis of FDA probably due to cytoplasmic changes effecting the enzymatic reaction. Also evident was an increase in loss of intracellular fluorescein and a decrease in PER polarization indicating that the cells have a decreased membrane integrity, possibly the result of an increased disorganization of the phospholipid hydrocarbon chains in the membrane interior. Experiments with PHA link the decreased membrane integrity with the eventual interphase death of the cells. In general it is shown that the fluorescent probes ANS, PER, and FDA provide useful ways to investigate order and microviscosity in the cell membrane surface and interior, membrane surface charges, internal membrane polarity changes, and membrane integrity. (U.S.)

Novel Tripod Amphiphiles for Membrane Protein Analysis

DEFF Research Database (Denmark)

Chae, Pil Seok; Kruse, Andrew C; Gotfryd, Kamil

2013-01-01

Integral membrane proteins play central roles in controlling the flow of information and molecules across membranes. Our understanding of membrane protein structures and functions, however, is seriously limited, mainly due to difficulties in handling and analysing these proteins in aqueous solution...

Two-step treatment of harmful industrial wastewater: an analysis of microbial reactor with integrated membrane retention for benzene and toluene removal

Directory of Open Access Journals (Sweden)

Trusek-Holownia Anna

2015-12-01

Full Text Available Standards for highly toxic and carcinogenic pollutants impose strict guidelines, requiring values close to zero, regarding the degradation of such pollutants in industrial streams. In many cases, classic bioremoval processes fail. Therefore, we proposed a stream leaving the microbial membrane bioreactor (MBR that is directed to an additional membrane separation mode (NF/RO. Under certain conditions, the integrated process not only benefits the environment but may also increase the profitability of the bioreactor operation. An appropriate model was developed and tested in which the bioremoval of benzene and toluene by Pseudomonas fluorescens was used as an example. This paper presents equations for selecting the operation parameters of the integrated system to achieve the expected degree of industrial wastewater purification.

Integrated Water Gas Shift Membrane Reactors Utilizing Novel, Non Precious Metal Mixed Matrix Membrane

Energy Technology Data Exchange (ETDEWEB)

Ferraris, John P. [Univ. of Texas-Dallas, Richardson, TX (United States). Dept. of Chemistry

2013-09-30

Nanoparticles of zeolitic imidazolate frameworks and other related hybrid materials were prepared by modifying published synthesis procedures by introducing bases, changing stoichiometric ratios, or adjusting reaction conditions. These materials were stable at temperatures >300 °C and were compatible with the polymer matrices used to prepare mixed- matrix membranes (MMMs). MMMs tested at 300 °C exhibited a >30 fold increase in permeability, compared to those measured at 35 °C, while maintaining H₂/CO₂ selectivity. Measurements at high pressure (up to 30 atm) and high temperature (up to 300 °C) resulted in an increase in gas flux across the membrane with retention of selectivity. No variations in permeability were observed at high pressures at either 35 or 300 °C. CO₂-induced plasticization was not observed for Matrimid®, VTEC, and PBI polymers or their MMMs at 30 atm and 300 °C. Membrane surface modification by cross-linking with ethanol diamine resulted in an increase in H₂/CO₂ selectivity at 35 °C. Spectrometric analysis showed that the cross-linking was effective to temperatures <150 °C. At higher temperatures, the cross-linked membranes exhibit a H₂/CO₂ selectivity similar to the uncross-linked polymer. Performance of the polybenzimidazole (PBI) hollow fibers prepared at Santa Fe Science and Technology (SFST, Inc.) showed increased flux and selectivity at 300 °C, which is comparable to a flat PBI membrane. A water-gas shift reactor has been built and currently being optimized for testing under DOE conditions.

Thermo-economic analysis of integrated membrane-SMR ITM-oxy-combustion hydrogen and power production plant

International Nuclear Information System (INIS)

Sanusi, Yinka S.; Mokheimer, Esmail M.A.; Habib, Mohamed A.

2017-01-01

Highlights: •A methane reforming reactor integrated to an oxy-combustion plant is proposed. •Co-production of power and hydrogen was investigated and presented. •Optimal thermo-economic operating conditions of the system were identified and presented. •The ion transport membrane oxygen separation unit has the highest capital cost. •The combustor has the highest exergy destruction. -- Abstract: The demand for hydrogen has greatly increased in the last decade due to the stringent regulations enacted to address environmental pollution concerns. Natural gas reforming is currently the most mature technology for large-scale hydrogen production. However, it is usually associated with greenhouse gas emissions. As part of the strategies to reduce greenhouse gas emissions, new designs need to be developed to integrate hydrogen production facilities that are based on natural gas reforming with carbon capture facilities. In this study, we carried out energy, exergy and economic analysis of hydrogen production in a steam methane reforming reactor integrated with an oxy-combustion plant for co-production of power and hydrogen. The results show that the overall system efficiency and hydrogen production efficiency monotonically increase with increasing the combustor exit temperature (CET), increasing the amount of hydrogen extracted and decreasing the auxiliary fuel added to the system. The optimal thermo-economic operating conditions of the system were obtained as reformer pressure of 15 bar, auxiliary fuel factor of 0.8 and hydrogen extraction factor of 0.6. The production cost of hydrogen using the proposed system, under these optimal operating conditions, is within the range suggested by the International Energy Agency (IEA). Further analysis shows that the capital cost of the membrane-air separation unit (ITM) has the major share in the total investment cost of the system and constitutes 37% of the total capital cost of the system at the CET of 1500 K. The exergy

The role of Slr0151, a tetratricopeptide repeat protein from Synechocystis sp. PCC 6803, during Photosystem II assembly and repair

Directory of Open Access Journals (Sweden)

Anna eRast

2016-05-01

Full Text Available The assembly and repair of photosystem II (PSII is facilitated by a variety of assembly factors. Among those, the tetratricopeptide repeat (TPR protein Slr0151 from Synechocystis sp. PCC 6803 (hereafter Synechocystis has previously been assigned a repair function under high light conditions (Yang et al., 2014, J. Integr. Plant Biol. 56, 1136-50. Here, we show that inactivation of Slr0151 affects thylakoid membrane ultrastructure even under normal light conditions. Moreover, the level and localization of Slr0151 are affected in a variety of PSII-related mutants. In particular, the data suggest a close functional relationship between Slr0151 and Sll0933, which interacts with Ycf48 during PSII assembly and is homologous to PAM68 in Arabidopsis thaliana. Immunofluorescence analysis revealed a punctate distribution of Slr0151 within several different membrane types in Synechocystis cells.

Surface modification of polysulfone membranes applied for a membrane reactor with immobilized alcohol dehydrogenase

DEFF Research Database (Denmark)

Hoffmann, Christian; Silau, Harald; Pinelo, Manuel

2018-01-01

activated by lithiation followed by functionalization with acid chlorides at 0 °C, permitting modification of commercial PSf membranes without compromising the mechanical integrity of the membrane. Post-functionalization polymer grafting was illustrated through both, a “grafting from” approach by surface...... initiated atom transfer radical polymerization (SI-ATRP) and by a “grafting to” approach exploiting Cu(I) catalyzed 1,3-cycloadditions of alkynes with azides (CuAAC) introducing hydrophilic polymers onto the membrane surface. Poly(1-vinyl imidazole) (pVim) grafted membranes were exploited as support...

Challege and Opportunities of Membrane Bioelctrochemical Reactors for Wastewater Treatment

OpenAIRE

Li, Jian

2016-01-01

Microbial fuel cells (MFCs) are potentially advantageous as an energy-efficient approach for wastewater treatment. Integrating membrane filtration with MFCs could be a viable option for advanced wastewater treatment with a low energy input. Such an integration is termed as membrane bioelectrochemical reactors (MBERs). Comparing to the conventional membrane bioreactors or anaerobic membrane bioreactors, MBER could be a competitive technology, due to the its advantages on energy consumption and...

Functional role of the extracellular N-terminal domain of neuropeptide Y subfamily receptors in membrane integration and agonist-stimulated internalization.

Science.gov (United States)

Lindner, Diana; Walther, Cornelia; Tennemann, Anja; Beck-Sickinger, Annette G

2009-01-01

The N terminus is the most variable element in G protein-coupled receptors (GPCRs), ranging from seven residues up to approximately 5900 residues. For family B and C GPCRs it is described that at least part of the ligand binding site is located within the N terminus. Here we investigated the role of the N terminus in the neuropeptide Y receptor family, which belongs to the class A of GPCRs. We cloned differentially truncated Y receptor mutants, in which the N terminus was partially or completely deleted. We found, that eight amino acids are sufficient for full ligand binding and signal transduction activity. Interestingly, we could show that no specific amino acids but rather the extension of the first transmembrane helix by any residues is sufficient for receptor activity but also for membrane integration in case of the hY(1) and the hY(4) receptors. In contrast, the complete deletion of the N terminus in the hY(2) receptors resulted in a mutant that is fully integrated in the membrane but does not bind the ligand very well and internalizes much slower compared to the wild type receptor. Interestingly, also these effects could be reverted by any N-terminal extension. Accordingly, the most important function of the N termini seems to be the stabilization of the first transmembrane helix to ensure the correct receptor structure, which obviously is essential for ligand binding, integration into the cell membrane and receptor internalization.

Ion transport through biological membranes an integrated theoretical approach

CERN Document Server

Mackey, Michael C

1975-01-01

This book illustrates some of the ways physics and mathematics have been, and are being, used to elucidate the underlying mechan­ isms of passive ion movement through biological membranes in general, and the membranes of excltable cells in particular. I have made no effort to be comprehensive in my introduction of biological material and the reader interested in a brief account of single cell electro­ physlology from a physically-oriented biologists viewpoint will find the chapters by Woodbury (1965) an excellent introduction. Part I is introductory in nature, exploring the basic electrical properties of inexcitable and excitable cell plasma membranes. Cable theory is utilized to illustrate the function of the non-decrementing action potential as a signaling mechanism for the long range trans­ mission of information in the nervous system, and to gain some in­ sight into the gross behaviour of neurons. The detailed analysis of Hodgkin and Huxley on the squid giant axon membrane ionic conductance properties...

Analysis of Protein-Membrane Interactions

DEFF Research Database (Denmark)

Kemmer, Gerdi Christine

Cellular membranes are complex structures, consisting of hundreds of different lipids and proteins. These membranes act as barriers between distinct environments, constituting hot spots for many essential functions of the cell, including signaling, energy conversion, and transport. These functions....... Discovered interactions were then probed on the level of the membrane using liposome-based assays. In the second part, a transmembrane protein was investigated. Assays to probe activity of the plasma membrane ATPase (Arabidopsis thaliana H+ -ATPase isoform 2 (AHA2)) in single liposomes using both giant...... are implemented by soluble proteins reversibly binding to, as well as by integral membrane proteins embedded in, cellular membranes. The activity and interaction of these proteins is furthermore modulated by the lipids of the membrane. Here, liposomes were used as model membrane systems to investigate...

Exergy analysis of the biogas sorption-enhanced chemical looping reforming process integrated with a high-temperature proton exchange membrane fuel cell

International Nuclear Information System (INIS)

Kasemanand, Sarunyou; Im-orb, Karittha; Tippawan, Phanicha; Wiyaratn, Wisitsree; Arpornwichanop, Amornchai

2017-01-01

Highlights: • A biogas reforming and fuel cell integrated process is considered. • Energy and exergy analyses of the integrated process are performed. • Increasing the nickel oxide-to-biogas ratio decreases the exergy efficiency. • The exergy destruction of the fuel cell increases with increasing cell temperature. • The exergy efficiency of the process is improved when heat integration is applied. - Abstract: A biogas sorption-enhanced chemical looping reforming process integrated with a high-temperature proton exchange membrane fuel cell is analyzed. Modeling of such an integrated process is performed by using a flowsheet simulator (Aspen plus). The exergy analysis is performed to evaluate the energy utilization efficiency of each unit and that of the integrated process. The effect of steam and nickel oxide to biogas ratios on the exergetic performance of the stand-alone biogas sorption-enhanced chemical looping reforming process is investigated. The total exergy destruction increases as the steam or nickel oxide to biogas ratio increases. The main exergy destruction is found at the air reactor. For the high-temperature proton exchange membrane fuel cell, the main exergy destruction is found at the cathode. The total exergy destruction increases when cell temperature increases, whereas the inverse effect is found when the current density is considered as a key parameter. Regarding the exergy efficiency, the results show opposite trend to the exergy destruction. The heat integration analysis is performed to improve the exergetic performance. It is found that the integrated process including the heat integration system can improve the exergy destruction and exergy efficiency of 48% and 60%, respectively.

Understanding Free Radicals: Isolating Active Thylakoid Membranes and Purifying the Cytochrome b6f Complex for Superoxide Generation Studies

Directory of Open Access Journals (Sweden)

Jason Stofleth

2012-01-01

Full Text Available All life persists in an environment that is rich in molecular oxygen. The production of oxygen free radicals, or superoxide, is a necessary consequence of the biogenesis of energy in cells. Both mitochondrial and photosynthetic electron transport chains have been found to produce superoxide associated with cell differentiation, proliferation, and cell death, thereby contributing to the effects of aging. Aerobic respiration in mitochondria consumes oxygen, whereas photosynthesis in chloroplasts or cyanobacteria produces oxygen. The increased concentration of molecular oxygen may serve to allow greater availability for the production of superoxide by cytochrome bc complexes in photosynthetic membranes compared to those of mitochondrial membranes. The isolation of well-coupled chloroplasts, containing the cytochrome b6f complex of oxygenic photosynthesis, is a vital initial step in the process of comparing the rate of production of superoxide to those of the homologous cytochrome bc1 complex of aerobic respiration. It is necessary to determine if the isolated chloroplasts have retained their oxygengenerating capability after isolation by an oxygen evolution assay with a Clark-type electrode. A necessary second step, which is the isolation of cytochrome b6f from spinach, has yet to be successfully performed. Oxygen measurements taken from chloroplasts in the presence of the uncoupler, NH4Cl, exhibited a rate of oxygen evolution over three times greater at 344 +/- 18 μmol O2/mg Chlorophyll a/hr than the rate of oxygen evolution without uncoupler at 109 +/- 29 μmol O2/mg Chlorophyll a/hr. These data demonstrate that the technique used to isolate spinach chloroplasts preserves their light-driven electron-transport activity, making them reliable for future superoxide assays.

Structure and physical properties of bio membranes and model membranes

International Nuclear Information System (INIS)

Tibor Hianik

2006-01-01

Bio membranes belong to the most important structures of the cell and the cell organelles. They play not only structural role of the barrier separating the external and internal part of the membrane but contain also various functional molecules, like receptors, ionic channels, carriers and enzymes. The cell membrane also preserves non-equilibrium state in a cell which is crucial for maintaining its excitability and other signaling functions. The growing interest to the bio membranes is also due to their unique physical properties. From physical point of view the bio membranes, that are composed of lipid bilayer into which are incorporated integral proteins and on their surface are anchored peripheral proteins and polysaccharides, represent liquid s crystal of smectic type. The bio membranes are characterized by anisotropy of structural and physical properties. The complex structure of bio membranes makes the study of their physical properties rather difficult. Therefore several model systems that mimic the structure of bio membranes were developed. Among them the lipid monolayers at an air-water interphase, bilayer lipid membranes, supported bilayer lipid membranes and liposomes are most known. This work is focused on the introduction into the physical word of the bio membranes and their models. After introduction to the membrane structure and the history of its establishment, the physical properties of the bio membranes and their models are stepwise presented. The most focus is on the properties of lipid monolayers, bilayer lipid membranes, supported bilayer lipid membranes and liposomes that were most detailed studied. This lecture has tutorial character that may be useful for undergraduate and graduate students in the area of biophysics, biochemistry, molecular biology and bioengineering, however it contains also original work of the author and his co-worker and PhD students, that may be useful also for specialists working in the field of bio membranes and model

Ultrastructural study on dynamics of lipid bodies and plastids during ripening of chili pepper fruits.

Science.gov (United States)

Liu, Lin

2013-03-01

Dynamics of lipid bodies and plastids in chili pepper fruits during ripening were investigated by means of transmission electron microscopy. Mesocarp of chili pepper fruits consists of collenchyma, normal parenchyma, and huge celled parenchyma. In mature green fruits, plastids contain numerous thylakoids that are well organized into grana in collenchyma, a strikingly huge amount of starch and irregularly organized thylakoids in normal parenchyma, and simple tubes rather than thylakoids in huge celled parenchyma. These morphological features suggest that plastids are chloroplasts in collenchyma, chloroamyloplasts in normal parenchyma, proplastids in huge celled parenchyma. As fruits ripen to red, plastids in all cell types convert to chromoplasts and, concomitantly, lipid bodies accumulate in both cytoplasm and chromoplasts. Cytosolic lipid bodies are lined up in a regular layer adjacent to plasma membrane. The cytosolic lipid body consists of a core surrounded by a membrane. The core is comprised of a more electron-dense central part enclosed by a slightly less electron-dense peripheral layer. Plastidial lipid bodies in collenchyma, normal parenchyma, and endodermis initiate as plastoglobuli, which in turn convert to rod-like structures. Therefore, plastidial lipid bodies are more dynamic than cytosolic lipid bodies. Both cytosolic and plastidial lipid bodies contain rich unsaturated lipids. Copyright © 2012 Elsevier Ltd. All rights reserved.

Integrating microbial fuel cells with anaerobic acidification and forward osmosis membrane for enhancing bio-electricity and water recovery from low-strength wastewater.

Science.gov (United States)

Liu, Jinmeng; Wang, Xinhua; Wang, Zhiwei; Lu, Yuqin; Li, Xiufen; Ren, Yueping

2017-03-01

Microbial fuel cells (MFCs) and forward osmosis (FO) are two emerging technologies with great potential for energy-efficient wastewater treatment. In this study, anaerobic acidification and FO membrane were simultaneously integrated into an air-cathode MFC (AAFO-MFC) for enhancing bio-electricity and water recovery from low-strength wastewater. During a long-term operation of approximately 40 days, the AAFO-MFC system achieved a continuous and relatively stable power generation, and the maximum power density reached 4.38 W/m 3 . The higher bio-electricity production in the AAFO-MFC system was mainly due to the accumulation of ethanol resulted from anaerobic acidification process and the rejection of FO membrane. In addition, a proper salinity environment in the system controlled by the addition of MF membrane enhanced the electricity production. Furthermore, the AAFO-MFC system produced a high quality effluent, with the removal rates of organic matters and total phosphorus of more than 97%. However, the nitrogen removal was limited for the lower rejection of FO membrane. The combined biofouling and inorganic fouling were responsible for the lower water flux of FO membrane, and the Desulfuromonas sp. utilized the ethanol for bio-electricity production was observed in the anode. These results substantially improve the prospects for simultaneous wastewater treatment and energy recovery, and further studies are needed to optimize the system integration and operating parameters. Copyright © 2016 Elsevier Ltd. All rights reserved.

Response of the photosynthetic system to altered protein composition and changes in environmental conditions

NARCIS (Netherlands)

Tóth, T.

2014-01-01

The photosynthetic thylakoid membrane has a hierarchically ordered structure containing pigment-protein complexes that capture solar radiation and convert it into chemical energy. Its highly dynamic structure is capable to continuously respond to the altered environmental conditions, e.g., light

Cyanobacterial photosystem II at 2.9-A resolution and the role of quinones, lipids, channels and chloride

NARCIS (Netherlands)

Guskov, Albert; Kern, Jan; Gabdulkhakov, Azat; Broser, Matthias; Zouni, Athina; Saenger, Wolfram

Photosystem II (PSII) is a large homodimeric protein-cofactor complex located in the photosynthetic thylakoid membrane that acts as light-driven water:plastoquinone oxidoreductase. The crystal structure of PSII from Thermosynechococcus elongatus at 2.9-A resolution allowed the unambiguous assignment

Mechanisms of energy transfer and conversion in plant Light-Harvesting Complex II

Energy Technology Data Exchange (ETDEWEB)

Barros, Tiago Ferreira de

2009-09-24

The light-harvesting complex of photosystem II (LHC-II) is the major antenna complex in plant photosynthesis. It accounts for roughly 30% of the total protein in plant chloroplasts, which makes it arguably the most abundant membrane protein on Earth, and binds about half of plant chlorophyll (Chl). The complex assembles as a trimer in the thylakoid membrane and binds a total of 54 pigment molecules, including 24 Chl a, 18 Chl b, 6 lutein (Lut), 3 neoxanthin (Neo) and 3 violaxanthin (Vio). LHC-II has five key roles in plant photosynthesis. It: (1) harvests sunlight and transmits excitation energy to the reaction centres of photosystems II and I, (2) regulates the amount of excitation energy reaching each of the two photosystems, (3) has a structural role in the architecture of the photosynthetic supercomplexes, (4) contributes to the tight appression of thylakoid membranes in chloroplast grana, and (5) protects the photosynthetic apparatus from photo damage by non photochemical quenching (NPQ). A major fraction of NPQ is accounted for its energy-dependent component qE. Despite being critical for plant survival and having been studied for decades, the exact details of how excess absorbed light energy is dissipated under qE conditions remain enigmatic. Today it is accepted that qE is regulated by the magnitude of the pH gradient ({delta}pH) across the thylakoid membrane. It is also well documented that the drop in pH in the thylakoid lumen during high-light conditions activates the enzyme violaxanthin de-epoxidase (VDE), which converts the carotenoid Vio into zeaxanthin (Zea) as part of the xanthophyll cycle. Additionally, studies with Arabidopsis mutants revealed that the photosystem II subunit PsbS is necessary for qE. How these physiological responses switch LHC-II from the active, energy transmitting to the quenched, energy-dissipating state, in which the solar energy is not transmitted to the photosystems but instead dissipated as heat, remains unclear and is the

Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers

International Nuclear Information System (INIS)

Mote, Kaustubh R.; Gopinath, T.; Traaseth, Nathaniel J.; Kitchen, Jason; Gor’kov, Peter L.; Brey, William W.; Veglia, Gianluigi

2011-01-01

Oriented solid-state NMR is the most direct methodology to obtain the orientation of membrane proteins with respect to the lipid bilayer. The method consists of measuring 1 H- 15 N dipolar couplings (DC) and 15 N anisotropic chemical shifts (CSA) for membrane proteins that are uniformly aligned with respect to the membrane bilayer. A significant advantage of this approach is that tilt and azimuthal (rotational) angles of the protein domains can be directly derived from analytical expression of DC and CSA values, or, alternatively, obtained by refining protein structures using these values as harmonic restraints in simulated annealing calculations. The Achilles’ heel of this approach is the lack of suitable experiments for sequential assignment of the amide resonances. In this Article, we present a new pulse sequence that integrates proton driven spin diffusion (PDSD) with sensitivity-enhanced PISEMA in a 3D experiment ([ 1 H, 15 N]-SE-PISEMA-PDSD). The incorporation of 2D 15 N/ 15 N spin diffusion experiments into this new 3D experiment leads to the complete and unambiguous assignment of the 15 N resonances. The feasibility of this approach is demonstrated for the membrane protein sarcolipin reconstituted in magnetically aligned lipid bicelles. Taken with low electric field probe technology, this approach will propel the determination of sequential assignment as well as structure and topology of larger integral membrane proteins in aligned lipid bilayers.

Further Improvement and System Integration of High Temperature Polymer Electrolyte Membrane Fuel Cells

DEFF Research Database (Denmark)

Li, Qingfeng; Jensen, Jens Oluf

The strategic developments of the FURIM are in three steps: (1) further improvement of the high temperature polymer membranes and related materials; (2) development of technological units including fuel cell stack, hydrocarbon reformer and afterburner, that are compatible with the HT-PEMFC; and (3......) integration of the HT-PEMFC stack with these compatible subunits. The main goal of the project is a 2kWel HT-PEMFC stack operating in a temperature range of 150-200°C, with a single cell performance target of 0.7 A/cm² at a cell voltage around 0.6 V. The target durability is more than 5,000 hours...

Osmotically and thermally isolated forward osmosis-membrane distillation (fo-md) integrated module for water treatment applications

KAUST Repository

Ghaffour, Noreddine

2016-09-01

An integrated forward osmosis-membrane distillation (FO-MD) module and systems and methods incorporating the module is disclosed providing higher efficiencies and using less energy. The FO-MD module is osmotically and thermally isolated. The isolation can prevent mixing of FO draw solution/FO permeate and MD feed, and minimize dilution of FO draw solution and cooling of MD feed. The module provides MD feed solution and FO draw solution streams that flow in the same module but are separated by an isolation barrier. The osmotically and thermally isolated FO-MD integrated module, systems and methods offer higher driving forces of both FO and MD processes, higher recovery, and wider application than previously proposed hybrid FO- MD systems.

Osmotically and thermally isolated forward osmosis-membrane distillation (fo-md) integrated module for water treatment applications

KAUST Repository

Ghaffour, NorEddine; Francis, Lijo; Li, Zhenyu; Valladares, Rodrigo; Alsaadi, Ahmad S.; Ghdaib, Muhannad Abu; Amy, Gary L.

2016-01-01

An integrated forward osmosis-membrane distillation (FO-MD) module and systems and methods incorporating the module is disclosed providing higher efficiencies and using less energy. The FO-MD module is osmotically and thermally isolated. The isolation can prevent mixing of FO draw solution/FO permeate and MD feed, and minimize dilution of FO draw solution and cooling of MD feed. The module provides MD feed solution and FO draw solution streams that flow in the same module but are separated by an isolation barrier. The osmotically and thermally isolated FO-MD integrated module, systems and methods offer higher driving forces of both FO and MD processes, higher recovery, and wider application than previously proposed hybrid FO- MD systems.

Regulation of multispanning membrane protein topology via post-translational annealing.

Science.gov (United States)

Van Lehn, Reid C; Zhang, Bin; Miller, Thomas F

2015-09-26

The canonical mechanism for multispanning membrane protein topogenesis suggests that protein topology is established during cotranslational membrane integration. However, this mechanism is inconsistent with the behavior of EmrE, a dual-topology protein for which the mutation of positively charged loop residues, even close to the C-terminus, leads to dramatic shifts in its topology. We use coarse-grained simulations to investigate the Sec-facilitated membrane integration of EmrE and its mutants on realistic biological timescales. This work reveals a mechanism for regulating membrane-protein topogenesis, in which initially misintegrated configurations of the proteins undergo post-translational annealing to reach fully integrated multispanning topologies. The energetic barriers associated with this post-translational annealing process enforce kinetic pathways that dictate the topology of the fully integrated proteins. The proposed mechanism agrees well with the experimentally observed features of EmrE topogenesis and provides a range of experimentally testable predictions regarding the effect of translocon mutations on membrane protein topogenesis.

Challenges in the Development of Functional Assays of Membrane Proteins

Directory of Open Access Journals (Sweden)

Sophie Demarche

2012-11-01

Full Text Available Lipid bilayers are natural barriers of biological cells and cellular compartments. Membrane proteins integrated in biological membranes enable vital cell functions such as signal transduction and the transport of ions or small molecules. In order to determine the activity of a protein of interest at defined conditions, the membrane protein has to be integrated into artificial lipid bilayers immobilized on a surface. For the fabrication of such biosensors expertise is required in material science, surface and analytical chemistry, molecular biology and biotechnology. Specifically, techniques are needed for structuring surfaces in the micro- and nanometer scale, chemical modification and analysis, lipid bilayer formation, protein expression, purification and solubilization, and most importantly, protein integration into engineered lipid bilayers. Electrochemical and optical methods are suitable to detect membrane activity-related signals. The importance of structural knowledge to understand membrane protein function is obvious. Presently only a few structures of membrane proteins are solved at atomic resolution. Functional assays together with known structures of individual membrane proteins will contribute to a better understanding of vital biological processes occurring at biological membranes. Such assays will be utilized in the discovery of drugs, since membrane proteins are major drug targets.

Functions of tocopherols in the cells of plants and other photosynthetic organisms.

Science.gov (United States)

Mokrosnop, V M

2014-01-01

Tocopherol synthesis has only been observed in photosynthetic organisms (plants, algae and some cyanobacteria). Tocopherol is synthesized in the inner membrane of chloroplasts and distributed between chloroplast membranes, thylakoids and plastoglobules. Physiological significance of tocopherols for human and animal is well-studied, but relatively little is known about their function in plant organisms. Among the best characterized functions oftocopherols in cells is their ability to scavenge and quench reactive oxygen species and fat-soluble by-products of oxidative stress. There are the data on the participation of different mechanisms of α-tocopherol action in protecting photosystem II (PS II) from photoinhibition both by deactivation of singlet oxygen produced by PSII and by reduction of proton permeability of thylakoid membranes, leading to acidification of lumen under high light conditions and activation of violaxanthin de-epoxidase. Additional biological activity of tocopherols, independent of its antioxidant functions have been demonstrated. Basic mechanisms for these effects are connected with the modulation of signal transduction pathways by specific tocopherols and, in some instances, by transcriptional activation of gene expression.

The outer membrane protein assembly machinery of Neisseria meningitidis

NARCIS (Netherlands)

Volokhina, E.B.|info:eu-repo/dai/nl/304837202

2009-01-01

Gram-negative bacteria are characterized by a cell envelope consisting of an inner membrane (IM) and an outer membrane (OM), which are separated by the peptidoglycan-containing periplasm. While the integral IM proteins are alpha-helical, all but one known integral OM proteins (OMPs) are

Motoneurons have different membrane resistance during fictive scratching and weight support

DEFF Research Database (Denmark)

Perreault, Marie-Claude

2002-01-01

locomotion; central pattern generator; spinal cord; synaptic integration; membrane conductance; glycine; postsynaptic inhibition; chloride......locomotion; central pattern generator; spinal cord; synaptic integration; membrane conductance; glycine; postsynaptic inhibition; chloride...

Recent Advances on Carbon Molecular Sieve Membranes (CMSMs and Reactors

Directory of Open Access Journals (Sweden)

Margot A. Llosa Tanco

2016-08-01

Full Text Available Carbon molecular sieve membranes (CMSMs are an important alternative for gas separation because of their ease of manufacture, high selectivity due to molecular sieve separation, and high permeance. The integration of separation by membranes and reaction in only one unit lead to a high degree of process integration/intensification, with associated benefits of increased energy, production efficiencies and reduced reactor or catalyst volume. This review focuses on recent advances in carbon molecular sieve membranes and their applications in membrane reactors.

Effects of High Hydrostatic Pressure on Escherichia coli Ultrastructure, Membrane Integrity and Molecular Composition as Assessed by FTIR Spectroscopy and Microscopic Imaging Techniques

Directory of Open Access Journals (Sweden)

María Prieto-Calvo

2014-12-01

Full Text Available High hydrostatic pressure (HHP is a novel food processing technology that is considered as an attractive alternative to conventional heat treatments for the preservation of foods, due to its lethal effects on pathogenic and spoilage microorganisms, while causing minor effects on food quality and sensorial attributes. This study is aimed at investigating how HHP treatments at varying intensities in the range 50–900 MPa affect the viability, membrane integrity, ultrastructure and molecular composition of Escherichia coli. Results of membrane integrity tests (measurement of cellular leakage and monitoring of propidium iodide uptake through fluorescence microscopy and ultrastructural observations by transmission electron microscopy demonstrated that HHP gave rise to cellular enlargement, membrane damage or detachment, DNA and protein denaturation and loss of intracellular contents. Fourier-transform infrared (FTIR spectroscopy analyses evidenced minor changes in molecular composition in response to high pressures, which were mostly observed on the spectral region w4 (1200–900 cm−1, mainly informative of carbohydrates and polysaccharides of the cell wall. These findings suggest that exposure of E. coli cells to HHP causes alterations in their physical integrity while producing minor modifications in biochemical cellular composition. The current study increases the knowledge on the mechanisms of E. coli inactivation by HHP and provides valuable information for the design of more effective food preservation regimes based on the integration of mild HHP in combination with other food preservation strategies into a multi-target hurdle technology approach.

Effects of high hydrostatic pressure on Escherichia coli ultrastructure, membrane integrity and molecular composition as assessed by FTIR spectroscopy and microscopic imaging techniques.

Science.gov (United States)

Prieto-Calvo, María; Prieto, Miguel; López, Mercedes; Alvarez-Ordóñez, Avelino

2014-12-18

High hydrostatic pressure (HHP) is a novel food processing technology that is considered as an attractive alternative to conventional heat treatments for the preservation of foods, due to its lethal effects on pathogenic and spoilage microorganisms, while causing minor effects on food quality and sensorial attributes. This study is aimed at investigating how HHP treatments at varying intensities in the range 50-900 MPa affect the viability, membrane integrity, ultrastructure and molecular composition of Escherichia coli. Results of membrane integrity tests (measurement of cellular leakage and monitoring of propidium iodide uptake through fluorescence microscopy) and ultrastructural observations by transmission electron microscopy demonstrated that HHP gave rise to cellular enlargement, membrane damage or detachment, DNA and protein denaturation and loss of intracellular contents. Fourier-transform infrared (FTIR) spectroscopy analyses evidenced minor changes in molecular composition in response to high pressures, which were mostly observed on the spectral region w4 (1200-900 cm-1), mainly informative of carbohydrates and polysaccharides of the cell wall. These findings suggest that exposure of E. coli cells to HHP causes alterations in their physical integrity while producing minor modifications in biochemical cellular composition. The current study increases the knowledge on the mechanisms of E. coli inactivation by HHP and provides valuable information for the design of more effective food preservation regimes based on the integration of mild HHP in combination with other food preservation strategies into a multi-target hurdle technology approach.

Modulation of photosystem II chlorophyll fluorescence by electrogenic events generated by photosystem I

NARCIS (Netherlands)

Bulychev, A.A.; Vredenberg, W.J.

2001-01-01

In an attempt to uncover electric field interactions between PS I and PS II during their functioning, fluorescence induction curves were measured on hydroxylamine-treated thylakoids of Chenopodium album under conditions ensuring low and high levels of photogenerated membrane potentials. In parallel

Contrasting effect of dark-chilling on chloroplast structure and arrangement of chlorophyll-protein complexes in pea and tomato : Plants with a different susceptibility to non-freezing temperature

NARCIS (Netherlands)

Garstka, Maciej; Venema, Jan Henk; Rumak, Izabela; Gieczewska, Katarzyna; Rosiak, Malgorzata; Koziol-Lipinska, Joanna; Kierdaszuk, Borys; Vredenberg, Wim J.; Mostowska, Agnieszka

2007-01-01

The effect of dark-chilling and subsequent photoactivation on chloroplast structure and arrangements of chlorophyll-protein complexes in thylakoid membranes was studied in chilling-tolerant (CT) pea and in chilling-sensitive (CS) tomato. Dark-chilling did not influence chlorophyll content and Chl

Contrasting effect of dark-chilling on chloroplast structure and arrangement of chlorophyll-protein complexes in pea and tomato: plants with a different susceptibility to non-freezing temperature

NARCIS (Netherlands)

Garstka, M.; Venema, J.H.; Rumak, I.; Gieczewska, K.; Rosiak, M.; Koziol-Lipinska, J.; Vredenberg, W.J.; Mostowska, A.

2007-01-01

The effect of dark-chilling and subsequent photoactivation on chloroplast structure and arrangements of chlorophyll-protein complexes in thylakoid membranes was studied in chilling-tolerant (CT) pea and in chilling-sensitive (CS) tomato. Dark-chilling did not influence chlorophyll content and Chl

An organelle-free assay for pea chloroplast Mg-chelatase: Resolution of the activity into soluble and membrane bound fractions

Energy Technology Data Exchange (ETDEWEB)

Walker, C.J.; Weinstein, J.D. (Clemson Univ, SC (United States))

1991-05-01

Mg-chelatase, which catalyzes the insertion of magnesium into protoporphyrin, lies at the branchpoint of heme and chlorophyll biosynthesis in chloroplasts. Since magnesium chelation is the first step unique to chlorophyll synthesis, one would expect this step to be highly regulated. However, to date little is known about the enzymology or regulation of Mg-chelatase due mostly to an inability to assay it's activity outside of the intact plastid. Here the authors report the first truly in vitro i.e. organelle-free, assay for Mg-chelatase. Mg-chelatase activity in intact pea chloroplasts which is 3 to 4 fold higher than in cucumber chloroplasts, survived chloroplast lysis and could be fractionated, by centrifugation, into supernatant and pellet components. Both of these fractions were required to reconstitute Mg-chelatase activity and both were inactivated by boiling; indicating that the enzyme is composed of soluble and membrane bound protein(s). The specific activity of the reconstituted system was typically 1 nmol Mg-Deuteroporphyrin/h/mg protein and activity was linear for at least 60 min under our assay conditions. ATP and magnesium were required for Mg-chelatase activity. The soluble component could be fractionated with ammonium sulfate. The product of the reaction was confirmed fluorometrically as the magnesium chelate of the porphyrin substrate. Crude separation of chloroplast membranes into thylakoids and envelopes, suggested that the membrane-bound component of Mg-chelatase is probably located in the envelope.

The Use of Green Leaf Membranes to Promote Appetite Control, Suppress Hedonic Hunger and Loose Body Weight.

Science.gov (United States)

Erlanson-Albertsson, Charlotte; Albertsson, Per-Åke

2015-09-01

On-going research aims at answering the question, which satiety signal is the most potent or which combination of satiety signals is the most potent to stop eating. There is also an aim at finding certain food items or food additives that could be used to specifically reduce food intake therapeutically. Therapeutic attempts to normalize body weight and glycaemia with single agents alone have generally been disappointing. The success of bariatric surgery illustrates the rationale of using several hormones to treat obesity and type-2-diabetes. We have found that certain components from green leaves, the thylakoids, when given orally have a similar rationale in inducing the release of several gut hormones at the same time. In this way satiety is promoted and hunger suppressed, leading to loss of body weight and body fat. The mechanism is a reduced rate of intestinal lipid hydrolysis, allowing the lipolytic products to reach the distal intestine and release satiety hormones. The thylakoids also regulate glucose uptake in the intestine and influences microbiota composition in the intestine in a prebiotic direction. Using thylakoids is a novel strategy for treatment and prevention of obesity.

Photosystem II Water Oxidation: Mechanism, Efficiency and Flux in Diverse Oxygenic Phototrophs

Energy Technology Data Exchange (ETDEWEB)

Dismukes, Gerard Charles [Rutgers Univ., Piscataway, NJ (United States); Ananyev, Gennady [Rutgers Univ., Piscataway, NJ (United States); Gates, Colin [Rutgers Univ., Piscataway, NJ (United States)

2018-01-09

In one year, we pursued four aims: 1) extend the VZAD model to allow analysis of PSII chlorophyll fluorescence emission as modulated by interaction with the WOC (partial success); 2) compare the solar energy conversion efficiencies of PSII-WOCs from intact cells, isolated thylakoid membranes and PSII core complexes and crystals from cyanobacterium Thermosynechococcus elongatus (collaboration with Lawrence Berkeley National Laboratory; some success after changing collaborator); 3) determine whether PSIIs can store light energy by pumping protons across the thylakoid membrane (PSII-cyclic electron flow) and how it is regulated within the green alga Chlorella ohadii (collaboration with the Hebrew University of Jerusalem; some success); and 4) genetically replace the native PSII-D1 protein subunit from a higher plant with two cyanobacterial D1 isoforms to test whether their functional advantages in growth and photoprotection can be transferred (collaboration with Rutgers University; success).

Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers.

Science.gov (United States)

Mote, Kaustubh R; Gopinath, T; Traaseth, Nathaniel J; Kitchen, Jason; Gor'kov, Peter L; Brey, William W; Veglia, Gianluigi

2011-11-01

Oriented solid-state NMR is the most direct methodology to obtain the orientation of membrane proteins with respect to the lipid bilayer. The method consists of measuring (1)H-(15)N dipolar couplings (DC) and (15)N anisotropic chemical shifts (CSA) for membrane proteins that are uniformly aligned with respect to the membrane bilayer. A significant advantage of this approach is that tilt and azimuthal (rotational) angles of the protein domains can be directly derived from analytical expression of DC and CSA values, or, alternatively, obtained by refining protein structures using these values as harmonic restraints in simulated annealing calculations. The Achilles' heel of this approach is the lack of suitable experiments for sequential assignment of the amide resonances. In this Article, we present a new pulse sequence that integrates proton driven spin diffusion (PDSD) with sensitivity-enhanced PISEMA in a 3D experiment ([(1)H,(15)N]-SE-PISEMA-PDSD). The incorporation of 2D (15)N/(15)N spin diffusion experiments into this new 3D experiment leads to the complete and unambiguous assignment of the (15)N resonances. The feasibility of this approach is demonstrated for the membrane protein sarcolipin reconstituted in magnetically aligned lipid bicelles. Taken with low electric field probe technology, this approach will propel the determination of sequential assignment as well as structure and topology of larger integral membrane proteins in aligned lipid bilayers. © Springer Science+Business Media B.V. 2011

Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling.

Science.gov (United States)

Zhang, T; Rawson, D M; Tosti, L; Carnevali, O

2008-04-01

This study investigated enzymatic activity of cathepsins and the membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling. Stage III oocytes (>0.5mm), obtained through dissection of anaesthetised female fish and desegregation of ovarian cumulus, were exposed to 2M methanol or 2M DMSO (both prepared in Hank's medium) for 30min at 22 degrees C before being loaded into 0.5ml plastic straws and placed into a programmable cooler. After controlled slow freezing, samples were plunged into liquid nitrogen (LN) and held for at least 10min, and thawed by immersing straws into a 27 degrees C water bath for 10s. Thawed oocytes were washed twice in Hank's medium. Cathepsin activity and membrane integrity of oocytes were assessed both after cryoprotectant treatment at 22 degrees C and after freezing in LN. Cathepsin B and L colorimetric analyses were performed using substrates Z-Arg-ArgNNap and Z-Phe-Arg-4MbetaNA-HCl, respectively, and 2-naphthylamine and 4-methoxy-2-naphthylamine were used as standards. Cathepsin D activity was performed by analysing the level of hydrolytic action on haemoglobin. Oocytes membrane integrity was assessed using 0.2% Trypan blue staining for 5min. Analysis of cathepsin activities showed that whilst the activity of cathepsin B and D was not affected by 2M DMSO treatment, their activity was lowered when treated with 2M methanol. Following freezing to -196 degrees C, the activity of all cathepsins (B, D and L) was significantly decreased in both 2M DMSO and 2M methanol. Trypan blue staining showed that 63.0+/-11.3% and 72.7+/-5.2% oocytes membrane stayed intact after DMSO and methanol treatment for 30min at 22 degrees C, respectively, whilst 14.9+/-2.6% and 1.4+/-0.8% stayed intact after freezing in DMSO and methanol to -196 degrees C. The results indicate that cryoprotectant treatment and freezing modified the activities of lysosomal enzymes involved in oocyte maturation and yolk

A recruiting protein of geranylgeranyl diphosphate synthase controls metabolic flux toward chlorophyll biosynthesis in rice.

Science.gov (United States)

Zhou, Fei; Wang, Cheng-Yuan; Gutensohn, Michael; Jiang, Ling; Zhang, Peng; Zhang, Dabing; Dudareva, Natalia; Lu, Shan

2017-06-27

In plants, geranylgeranyl diphosphate (GGPP) is produced by plastidic GGPP synthase (GGPPS) and serves as a precursor for vital metabolic branches, including chlorophyll, carotenoid, and gibberellin biosynthesis. However, molecular mechanisms regulating GGPP allocation among these biosynthetic pathways localized in the same subcellular compartment are largely unknown. We found that rice contains only one functionally active GGPPS, OsGGPPS1, in chloroplasts. A functionally active homodimeric enzyme composed of two OsGGPPS1 subunits is located in the stroma. In thylakoid membranes, however, the GGPPS activity resides in a heterodimeric enzyme composed of one OsGGPPS1 subunit and GGPPS recruiting protein (OsGRP). OsGRP is structurally most similar to members of the geranyl diphosphate synthase small subunit type II subfamily. In contrast to members of this subfamily, OsGRP enhances OsGGPPS1 catalytic efficiency and specificity of GGPP production on interaction with OsGGPPS1. Structural biology and protein interaction analyses demonstrate that affinity between OsGRP and OsGGPPS1 is stronger than between two OsGGPPS1 molecules in homodimers. OsGRP determines OsGGPPS1 suborganellar localization and directs it to a large protein complex in thylakoid membranes, consisting of geranylgeranyl reductase (OsGGR), light-harvesting-like protein 3 (OsLIL3), protochlorophyllide oxidoreductase (OsPORB), and chlorophyll synthase (OsCHLG). Taken together, genetic and biochemical analyses suggest OsGRP functions in recruiting OsGGPPS1 from the stroma toward thylakoid membranes, thus providing a mechanism to control GGPP flux toward chlorophyll biosynthesis.

Palladium based membranes and membrane reactors for hydrogen production and purification : An overview of research activities at Tecnalia and TU/e

NARCIS (Netherlands)

Fernandez, E.; Helmi Siasi Farimani, A.; Medrano Jimenez, J.A.; Coenen, K.T.; Arratibel Plazaola, A.; Melendez Rey, J.; de Nooijer, N.C.A.; Viviente, J.L.; Zuniga, J.; van Sint Annaland, M.; Gallucci, F.; Pacheco Tanaka, D.A.

2017-01-01

In this paper, the main achievements of several European research projects on Pd based membranes and Pd membrane reactors for hydrogen production are reported. Pd-based membranes have received an increasing interest for separation and purification of hydrogen. In addition, the integration of such

Ceramic membranes for gas processing in coal gasification

Energy Technology Data Exchange (ETDEWEB)

Smart, S.; Lin, C.X.C.; Ding, L.; Thambimuthu, K.; da Costa, J.C.D. [University of Queensland, Brisbane, Qld. (Australia)

2010-07-01

Pre-combustion options via coal gasification, especially integrated gasification combined cycle (IGCC) processes, are attracting the attention of governments, industry and the research community as an attractive alternative to conventional power generation. It is possible to build an IGCC plant with CCS with conventional technologies however; these processes are energy intensive and likely to reduce power plant efficiencies. Novel ceramic membrane technologies, in particular molecular sieving silica (MSS) and pervoskite membranes, offer the opportunity to reduce efficiency losses by separating gases at high temperatures and pressures. MSS membranes can be made preferentially selective for H{sub 2}, enabling both enhanced production, via a water-gas shift membrane reactor, and recovery of H{sub 2} from the syngas stream at high temperatures. They also allow CO{sub 2} to be concentrated at high pressures, reducing the compression loads for transportation and enabling simple integration with CO{sub 2} storage or sequestration operations. Perovskite membranes provide a viable alternative to cryogenic distillation for air separation by delivering the tonnage of oxygen required for coal gasification at a reduced cost. In this review we examine ceramic membrane technologies for high temperature gas separation and discuss the operational, mechanical, design and process considerations necessary for their successful integration into IGCC with CCS systems.

Intelligent Membranes: Dream or Reality?

Directory of Open Access Journals (Sweden)

Annarosa Gugliuzza

2013-07-01

Full Text Available Intelligent materials are claimed to overcome current drawbacks associated with the attainment of high standards of life, health, security and defense. Membrane-based sensors represent a category of smart systems capable of providing a large number of benefits to different markets of textiles, biomedicine, environment, chemistry, agriculture, architecture, transport and energy. Intelligent membranes can be characterized by superior sensitivity, broader dynamic range and highly sophisticated mechanisms of autorecovery. These prerogatives are regarded as the result of multi-compartment arrays, where complementary functions can be accommodated and well-integrated. Based on the mechanism of “sense to act”, stimuli-responsive membranes adapt themselves to surrounding environments, producing desired effects such as smart regulation of transport, wetting, transcription, hydrodynamics, separation, and chemical or energy conversion. Hopefully, the design of new smart devices easier to manufacture and assemble can be realized through the integration of sensing membranes with wireless networks, looking at the ambitious challenge to establish long-distance communications. Thus, the transfer of signals to collecting systems could allow continuous and real-time monitoring of data, events and/or processes.

On the potential application of polar and temperate marine microalgae for EPA and DHA production

NARCIS (Netherlands)

Boelen, P.; van Dijk, R.; Sinninghe Damsté, J.S.; Rijpstra, W.I.C.; Buma, A.G.J.

2013-01-01

Long chain polyunsaturated fatty acids (LC-PUFAs) such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are considered essential omega-3 fatty acids in human nutrition. In marine microalgae EPA and/or DHA are allegedly involved in the regulation of membrane fluidity and thylakoid

Activity ranking of synthetic analogs targeting vascular endothelial growth factor receptor 2 by an integrated cell membrane chromatography system.

Science.gov (United States)

Wang, Dongyao; Lv, Diya; Chen, Xiaofei; Liu, Yue; Ding, Xuan; Jia, Dan; Chen, Langdong; Zhu, Zhenyu; Cao, Yan; Chai, Yifeng

2015-12-01

Evaluating the biological activities of small molecules represents an important part of the drug discovery process. Cell membrane chromatography (CMC) is a well-developed biological chromatographic technique. In this study, we have developed combined SMMC-7721/CMC and HepG2/CMC with high-performance liquid chromatography and time-of-flight mass spectrometry to establish an integrated screening platform. These systems was subsequently validated and used for evaluating the activity of quinazoline compounds, which were designed and synthesized to target vascular endothelial growth factor receptor 2. The inhibitory activities of these compounds towards this receptor were also tested using a classical caliper mobility shift assay. The results revealed a significant correlation between these two methods (R(2) = 0.9565 or 0.9420) for evaluating the activities of these compounds. Compared with traditional methods of evaluating the activities analogous compounds, this integrated cell membrane chromatography screening system took less time and was more cost effective, indicating that it could be used as a practical method in drug discovery. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Factors Determining the Oxygen Permeability of Biological Membranes: Oxygen Transport Across Eye Lens Fiber-Cell Plasma Membranes.

Science.gov (United States)

Subczynski, Witold Karol; Widomska, Justyna; Mainali, Laxman

2017-01-01

Electron paramagnetic resonance (EPR) spin-label oximetry allows the oxygen permeability coefficient to be evaluated across homogeneous lipid bilayer membranes and, in some cases, across coexisting membrane domains without their physical separation. The most pronounced effect on oxygen permeability is observed for cholesterol, which additionally induces the formation of membrane domains. In intact biological membranes, integral proteins induce the formation of boundary and trapped lipid domains with a low oxygen permeability. The effective oxygen permeability coefficient across the intact biological membrane is affected not only by the oxygen permeability coefficients evaluated for each lipid domain but also by the surface area occupied by these domains in the membrane. All these factors observed in fiber cell plasma membranes of clear human eye lenses are reviewed here.

Photothermal IR spectroscopy with perforated membrane micromechanical resonators

DEFF Research Database (Denmark)

Kurek, Maksymilian

-IR method. In order to overcome them, string resonators were replaced by membranes. A reliable sampling technique was maintained by adding perforation to membranes and thereby essentially getting membrane porous filters. Membranes gave also access to fully integrated magnetic transduction that allowed...... for significant shrinkage and simplification of the system. An analytical model of a locally heated membrane was developed and confirmed through FEM simulations. Then, low stress silicon nitride perforated membranes were fabricated and characterized using two different experimental setups that employed optical...

A Femtosecond Visible/Visible and Visible/Mid-Infrared Transient Absorption Study of the Light Harvesting Complex II

NARCIS (Netherlands)

Stahl, A.D.; Di Donato, M.; van Stokkum, I.H.M.; van Grondelle, R.; Groot, M.L.

2009-01-01

Light harvesting complex II (LHCII) is the most abundant protein in the thylakoid membrane of higher plants and green algae. LHCII acts to collect solar radiation, transferring this energy mainly toward photosystem II, with a smaller amount going to photosystem I; it is then converted into a

Architecture of the light-harvesting apparatus of the eustigmatophyte alga Nannochloropsis oceanica

Czech Academy of Sciences Publication Activity Database

Litvín, Radek; Bína, David; Herbstová, Miroslava; Gardian, Zdenko

2016-01-01

Roč. 130, 1-3 (2016), s. 137-150 ISSN 0166-8595 R&D Projects: GA ČR(CZ) GP14-01377P Institutional support: RVO:60077344 Keywords : Light harvesting * Thylakoid membrane * Vaucheriaxanthin * Violaxanthin–chlorophyll protein Subject RIV: BO - Biophysics Impact factor: 3.864, year: 2016

Integrated membrane and microbial fuel cell technologies for enabling energy-efficient effluent Re-use in power plants.

Science.gov (United States)

Shrestha, Namita; Chilkoor, Govinda; Xia, Lichao; Alvarado, Catalina; Kilduff, James E; Keating, John J; Belfort, Georges; Gadhamshetty, Venkataramana

2017-06-15

Municipal wastewater is an attractive alternative to freshwater sources to meet the cooling water needs of thermal power plants. Here we offer an energy-efficient integrated microbial fuel cell (MFC)/ultrafiltration (UF) process to purify primary clarifier effluent from a municipal wastewater treatment plant for use as cooling water. The microbial fuel cell was shown to significantly reduce chemical oxygen demand (COD) in the primary settled wastewater effluent upstream of the UF module, while eliminating the energy demand required to deliver dissolved oxygen in conventional aerobic treatment. We investigated surface modification of the UF membranes to control fouling. Two promising hydrophilic monomers were identified in a high-throughput search: zwitterion (2-(Methacryloyloxy)-ethyl-dimethyl-(3-sulfopropyl ammoniumhydroxide, abbreviated BET SO 3 - ), and amine (2-(Methacryloyloxy) ethyl trimethylammonium chloride, abbreviated N(CH 3 ) 3 + ). Monomers were grafted using UV-induced polymerization on commercial poly (ether sulfone) membranes. Filtration of MFC effluent by membranes modified with BET SO 3 - and N(CH 3 ) 3 + exhibited a lower rate of resistance increase and lower energy consumption than the commercially available membrane. The MFC/UF process produced high quality cooling water that meets the Electrical Power Research Institute (EPRI) recommendations for COD, a suite of metals (Fe, Al, Cu, Zn, Si, Mn, S, Ca and Mg), and offered extremely low corrosion rates (<0.05 mm/yr). A series of AC and DC diagnostic tests were used to evaluate the MFC performance. Copyright © 2017 Elsevier Ltd. All rights reserved.

Combined effects of simulated acid rain and lanthanum chloride on chloroplast structure and functional elements in rice.

Science.gov (United States)

Hu, Huiqing; Wang, Lihong; Zhou, Qing; Huang, Xiaohua

2016-05-01

Acid rain and rare earth element (REE) pollution exist simultaneously in many agricultural regions. However, how REE pollution and acid rain affect plant growth in combination remains largely unknown. In this study, the combined effects of simulated acid rain and lanthanum chloride (LaCl3) on chloroplast morphology, chloroplast ultrastructure, functional element contents, chlorophyll content, and the net photosynthetic rate (P n) in rice (Oryza sativa) were investigated by simulating acid rain and rare earth pollution. Under the combined treatment of simulated acid rain at pH 4.5 and 0.08 mM LaCl3, the chloroplast membrane was smooth, proteins on this membrane were uniform, chloroplast structure was integrated, and the thylakoids were orderly arranged, and simulated acid rain and LaCl3 exhibited a mild antagonistic effect; the Mg, Ca, Mn contents, the chlorophyll content, and the P n increased under this combined treatment, with a synergistic effect of simulated acid rain and LaCl3. Under other combined treatments of simulated acid rain and LaCl3, the chloroplast membrane surface was uneven, a clear "hole" was observed on the surface of chloroplasts, and the thylakoids were dissolved and loose; and the P n and contents of functional elements (P, Mg, K, Ca, Mn, Fe, Ni, Cu, Zn and Mo) and chlorophyll decreased. Under these combined treatments, simulated acid rain and LaCl3 exhibited a synergistic effect. Based on the above results, a model of the combined effects of simulated acid rain and LaCl3 on plant photosynthesis was established in order to reveal the combined effects on plant photosynthesis, especially on the photosynthetic organelle-chloroplast. Our results would provide some references for further understanding the mechanism of the combined effects of simulated acid rain and LaCl3 on plant photosynthesis.

Lysosomal enlargement and lysosomal membrane destabilisation in mussel digestive cells measured by an integrative index

International Nuclear Information System (INIS)

Izagirre, Urtzi; Marigomez, Ionan

2009-01-01

Lysosomal responses (enlargement and membrane destabilisation) in mussel digestive cells are well-known environmental stress biomarkers in pollution effects monitoring in marine ecosystems. Presently, in laboratory and field studies, both responses were measured separately (in terms of lysosomal volume density - Vv - and labilisation period -LP) and combined (lysosomal response index - LRI) in order to contribute to their understanding and to develop an index useful for decisions makers. LRI integrates Vv and LP, which are not necessarily dependent lysosomal responses. It is unbiased and more sensitive than Vv and LP alone and diminishes background due to confounding factors. LRI provides a simple numerical index (consensus reference = 0; critical threshold = 1) directly related to the pollution impact degree. Moreover, LRI can be represented in a way that allows the interpretation of lysosomal responses, which is useful for environmental scientists. - Lysosomal responses to pollutants measured by an integrative index.

A novel gas separation integrated membrane bioreactor to evaluate the impact of self-generated biogas recycling on continuous hydrogen fermentation

International Nuclear Information System (INIS)

Bakonyi, Péter; Buitrón, Germán; Valdez-Vazquez, Idania; Nemestóthy, Nándor; Bélafi-Bakó, Katalin

2017-01-01

Highlights: • A Gas Separation Membrane Bioreactor was designed to improve H_2 production. • Headspace gas after enrichment by PDMS membranes was used for reactor sparging. • Stripping the bioreactor with a CO_2-enriched gas enhanced the H_2 fermentation. - Abstract: A Gas Separation Membrane Bioreactor (GSMBR) by integrating membrane technology with a continuous biohydrogen fermenter was designed. The feasibility of this novel configuration for the improvement of hydrogen production capacity was tested by stripping the fermentation liquor with CO_2- and H_2-enriched gases, obtained directly from the bioreactor headspace. The results indicated that sparging the bioreactor with the CO_2-concentrated fraction of the membrane separation unit (consisting of two PDMS modules) enhanced the steady-state H_2 productivity (8.9–9.2 L H_2/L-d) compared to the membrane-less control CSTR to be characterized with 6.96–7.35 L H_2/L-d values. On the other hand, purging with the H_2-rich gas strongly depressed the achievable productivity (2.7–3.03 L H_2/L-d). Microbial community structure and soluble metabolic products were monitored to assess the GSMBR behavior. The study demonstrated that stripping the bioH_2 fermenter with its own, self-generated atmosphere after adjusting its composition (to higher CO_2-content) can be a promising way to intensify dark fermentative H_2 evolution.

Micro direct methanol fuel cell with perforated silicon-plate integrated ionomer membrane

DEFF Research Database (Denmark)

Larsen, Jackie Vincent; Dalslet, Bjarke Thomas; Johansson, Anne-Charlotte Elisabeth Birgitta

2014-01-01

This article describes the fabrication and characterization of a silicon based micro direct methanol fuel cell using a Nafion ionomer membrane integrated into a perforated silicon plate. The focus of this work is to provide a platform for micro- and nanostructuring of a combined current collector...... at a perforation ratio of 40.3%. The presented fuel cells also show a high volumetric peak power density of 2 mW cm−3 in light of the small system volume of 480 μL, while being fully self contained and passively feed....... and catalytic electrode. AC impedance spectroscopy is utilized alongside IV characterization to determine the influence of the plate perforation geometries on the cell performance. It is found that higher ratios of perforation increases peak power density, with the highest achieved being 2.5 mW cm−2...

Membrane Technologies in Wine Industry: An Overview.

Science.gov (United States)

El Rayess, Youssef; Mietton-Peuchot, Martine

2016-09-09

Membrane processes are increasingly reported for various applications in wine industry such as microfiltration, electrodialysis, and reverse osmosis, but also emerging processes as bipolar electrodialysis and membrane contactor. Membrane-based processes are playing a critical role in the field of separation/purification, clarification, stabilization, concentration, and de-alcoholization of wine products. They begin to be an integral part of the winemaking process. This review will provide an overview of recent developments, applications, and published literature in membrane technologies applied in wine industry.

Mapping of unfolding states of integral helical membrane proteins by GPS-NMR and scattering techniques

DEFF Research Database (Denmark)

Calcutta, Antonello; Jessen, Christian M; Behrens, Manja Annette

2012-01-01

induced by unfolding of an integral membrane protein, namely TFE-induced unfolding of KcsA solubilized by the n-dodecyl ß-d-maltoside (DDM) surfactant is investigated by the recently introduced GPS-NMR (Global Protein folding State mapping by multivariate NMR) (Malmendal et al., PlosONE 5, e10262 (2010......)) along with dynamic light scattering (DLS) and small-angle X-ray scattering (SAXS). GPS-NMR is used as a tool for fast analysis of the protein unfolding processes upon external perturbation, and DLS and SAXS are used for further structural characterization of the unfolding states. The combination allows...

Enhancing Membrane Protein Identification Using a Simplified Centrifugation and Detergent-Based Membrane Extraction Approach.

Science.gov (United States)

Zhou, Yanting; Gao, Jing; Zhu, Hongwen; Xu, Jingjing; He, Han; Gu, Lei; Wang, Hui; Chen, Jie; Ma, Danjun; Zhou, Hu; Zheng, Jing

2018-02-20

Membrane proteins may act as transporters, receptors, enzymes, and adhesion-anchors, accounting for nearly 70% of pharmaceutical drug targets. Difficulties in efficient enrichment, extraction, and solubilization still exist because of their relatively low abundance and poor solubility. A simplified membrane protein extraction approach with advantages of user-friendly sample processing procedures, good repeatability and significant effectiveness was developed in the current research for enhancing enrichment and identification of membrane proteins. This approach combining centrifugation and detergent along with LC-MS/MS successfully identified higher proportion of membrane proteins, integral proteins and transmembrane proteins in membrane fraction (76.6%, 48.1%, and 40.6%) than in total cell lysate (41.6%, 16.4%, and 13.5%), respectively. Moreover, our method tended to capture membrane proteins with high degree of hydrophobicity and number of transmembrane domains as 486 out of 2106 (23.0%) had GRAVY > 0 in membrane fraction, 488 out of 2106 (23.1%) had TMs ≥ 2. It also provided for improved identification of membrane proteins as more than 60.6% of the commonly identified membrane proteins in two cell samples were better identified in membrane fraction with higher sequence coverage. Data are available via ProteomeXchange with identifier PXD008456.

Stromal serine protein kinase activity in spinach chloroplasts

International Nuclear Information System (INIS)

Cortez, N.; Lucero, H.A.; Vallejos, R.H.

1987-01-01

At least twelve 32 P-labeled stromal proteins were detected by electrophoresis under denaturing conditions when intact chloroplasts were incubated with 32 Pi, in the light but only three were detected in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) or in the dark. Incubation of isolated stroma with [gamma- 32 P]ATP resulted in the preferential phosphorylation of one of them, a 70-kDa polypeptide, in serine residues. Thylakoid membranes in the dark promoted the phosphorylation of two additional stromal polypeptides of 55 and 40 kDa. Illumination during the phosphorylation of stroma in the presence of thylakoids stimulated severalfold the labeling of the 40-kDa polypeptide but not when DCMU was added. The protein kinase activity present in isolated stroma phosphorylated exogenous substrates like histone III, phosvitin, histone II, and casein with specific activities of 3, 1.8, 0.7, and 0.2 pmol X mg-1 X min-1. Histone III polypeptides were phosphorylated differently by stroma and by thylakoids in the dark. Moreover, histone III phosphorylated by thylakoids in the dark yielded a pattern of phosphopeptides after V8 protease treatment that was different from the pattern obtained when histone III was phosphorylated by stroma

Effects of hepatitis B virus S protein exposure on sperm membrane integrity and functions.

Directory of Open Access Journals (Sweden)

XiangJin Kang

Full Text Available BACKGROUND: Hepatitis B is a public health problem worldwide. Viral infection can affect a man's fertility, but only scant information about the influence of hepatitis B virus (HBV infection on sperm quality is available. The purpose of this study was to investigate the effect of hepatitis B virus S protein (HBs on human sperm membrane integrity and functions. METHODS/PRINCIPAL FINDINGS: Reactive oxygen species (ROS, lipid peroxidation (LP, total antioxidant capacity (TAC and phosphatidylserine (PS externalization were determined. The terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL assays and flow cytometric analyses were performed. (1 After 3 h incubation with 25 µg/ml of HBs, the average rates of ROS positive cells, annexin V-positive/propidium iodide (PI-negative cells, Caspases-3,-8,-9 positive cells and TUNEL-positive cells were significantly increased in the test groups as compared to those in the control groups, while TAC level was decreased when compared with the control. The level of malondialdehyde (MDA in the sperm cells exposed to 50 µg/ml of HBs for 3 h was significantly higher than that in the control (P<0.05-0.01. (2 HBs increased the MDA levels and the numbers of ROS positive cells, annexin V-positive/PI-negative cells, caspases-3, -8, -9 positive cells and TUNEL-positive cells in a dose-dependent manner. (3 HBs monoclonal antibody (MAb and N-Acetylcysteine (NAC reduced the number of ROS-positive sperm cells. (4 HBs decreased the TAC levels in sperm cells in a dose-dependent manner. CONCLUSION: HBs exposure could lead to ROS generation, lipid peroxidation, TAC reduction, PS externalization, activation of caspases, and DNA fragmentation, resulting in increased apoptosis of sperm cells and loss of sperm membrane integrity and causing sperm dysfunctions.

Structure, function and regulation of plant photosystem I

NARCIS (Netherlands)

Jensen, Poul Erik; Bassi, Roberto; Boekema, Egbert J.; Dekker, Jan P.; Jansson, Stefan; Leister, Dario; Robinson, Colin; Scheller, Henrik Vibe

Photosystem I (PSI) is a multisubunit protein complex located in the thylakoid membranes of green plants and algae, where it initiates one of the first steps of solar energy conversion by light-driven electron transport. In this review, we discuss recent progress on several topics related to the

Structure, function and regulation of plant photosystem I

NARCIS (Netherlands)

Jensen, P.E.; Bassi, R.; Boekema, E.J.; Dekker, J.P.; Jansson, S.; Leister, D.; Robinson, C.; Scheller, H.V.

2007-01-01

Photosystem I (PSI) is a multisubunit protein complex located in the thylakoid membranes of green plants and algae, where it initiates one of the first steps of solar energy conversion by light-driven electron transport. In this review, we discuss recent progress on several topics related to the

Tissue integrity is essential for ectopic implantation of human endometrium in the chicken chorioallantoic membrane.

Science.gov (United States)

Nap, Annemiek W; Groothuis, Patrick G; Demir, Ayse Y; Maas, Jacques W M; Dunselman, Gerard A J; de Goeij, Anton F P M; Evers, Johannes L H

2003-01-01

Not all women with patent tubes develop clinically manifest endometriosis. Quality and quantity of endometrium in retrograde menstruation may be the determining factor in the development of the disease. We hypothesize that retrograde shedding of endometrial fragments with preserved integrity facilitates implantation of endometrium in ectopic locations, resulting in endometriotic lesion development. We evaluate the impact of tissue integrity on the success of endometriosis-like lesion development in the chicken embryo chorioallantoic membrane (CAM) model. Menstrual and non-menstrual (cyclic) endometrium were collected by biopsy, and either minced or enzymatically dispersed. Spontaneously shed menstrual effluent was collected by a menstrual cup, and cells and tissue were isolated. We evaluated whether infiltration or lesion formation in the CAM occurred after transplantation of endometrium onto the CAM. Transplantation of biopsied menstrual and cyclic endometrium fragments, and of endometrium fragments >1 mm(3) isolated from menstrual effluent, resulted in lesion formation. Transplantation of endometrial cells isolated from menstrual effluent did not lead to lesion formation. After transplantation of digested biopsied cyclic endometrium, infiltration in the CAM but no lesions were observed. In the CAM assay, integrity of tissue architecture determines success of implantation of human endometrium in ectopic locations.

Combination of Electrochemical Processes with Membrane Bioreactors for Wastewater Treatment and Fouling Control: A Review

OpenAIRE

Ensano, Benny M. B.; Borea, Laura; Naddeo, Vincenzo; Belgiorno, Vincenzo; de Luna, Mark D. G.; Ballesteros, Florencio C.

2016-01-01

This paper provides a critical review about the integration of electrochemical processes into membrane bioreactors (MBR) in order to understand the influence of these processes on wastewater treatment performance and membrane fouling control. The integration can be realized either in an internal or an external configuration. Electrically enhanced membrane bioreactors or electro membrane bioreactors (eMBRs) combine biodegradation, electrochemical and membrane filtration processes into one syst...

Combination of electrochemical processes with membrane bioreactors for wastewater treatment and fouling control: A review

OpenAIRE

Benny Marie B. Ensano; Laura Borea; Vincenzo Naddeo; Vincenzo Belgiorno; Mark Daniel G. de Luna; Mark Daniel G. de Luna; Florencio C. Ballesteros, Jr.; Florencio C. Ballesteros, Jr.

2016-01-01

This paper provides a critical review about the integration of electrochemical processes into membrane bioreactors (MBR) in order to understand the influence of these processes on wastewater treatment performance and membrane fouling control. The integration can be realized either in an internal or an external configuration. Electrically enhanced membrane bioreactors or electro membrane bioreactors (eMBRs) combine biodegradation, electrochemical and membrane filtration processes into one syst...

Protein kinase that phosphorylates light-harvesting complex is autophosphorylated and is associated with photosystem II

International Nuclear Information System (INIS)

Coughlan, S.J.; Hind, G.

1987-01-01

Thylakoid membranes were phosphorylated with [γ- 32 P]ATP and extracted with octyl glucoside and cholate. Among the radiolabeled phosphoproteins in the extract was a previously characterized protein kinase of 64-kDa apparent mass. The ability of this enzyme to undergo autophosphorylation in situ was used to monitor its distribution in the membrane. Fractionation studies showed that the kinase is confined to granal regions of the thylakoid, where it appears to be associated with the light-harvesting chlorophyll-protein complex of photosystem II. The kinetics of kinase autophosphorylation were investigated both in situ and in extracted, purified enzyme. In the membrane, autophosphorylation saturated within 20-30 min and was reversed with a half-time of 7-8 min upon removal of ATP or oxidative inactivation of the kinase; the accompanying dephosphorylation of light-harvesting complex was slower and kinetically complex. Fluoride (10 mM) inhibited these dephosphorylations. Autophosphorylation of the isolated kinase was independent of enzyme concentration, indicative of an intramolecular mechanism. A maximum of one serine residue per mole of kinase was esterified. Autophosphorylation was more rapid in the presence of histone IIIs, an exogenous substrate. Dephosphorylation of the isolated enzyme was not observed

Transfer of the cytochrome P450-dependent dhurrin pathway from Sorghum bicolor into Nicotiana tabacum chloroplasts for light-driven synthesis

DEFF Research Database (Denmark)

Gnanasekaran, Thiyagarajan; Karcher, Daniel; Nielsen, Agnieszka Janina Zygadlo

2016-01-01

. For this purpose, we stably engineered the dhurrin pathway from Sorghum bicolor into the chloroplasts of Nicotiana tabacum (tobacco). Dhurrin is a cyanogenic glucoside and its synthesis from the amino acid tyrosine is catalysed by two membrane-bound cytochrome P450 enzymes (CYP79A1 and CYP71E1) and a soluble...... glucosyltransferase (UGT85B1), and is dependent on electron transfer from a P450 oxidoreductase. The entire pathway was introduced into the chloroplast by integrating CYP79A1, CYP71E1, and UGT85B1 into a neutral site of the N. tabacum chloroplast genome. The two P450s and the UGT85B1 were functional when expressed...... compared to 6% in sorghum. The results obtained pave the way for plant P450s involved in the synthesis of economically important compounds to be engineered into the thylakoid membrane of chloroplasts, and demonstrate that their full catalytic cycle can be driven directly by photosynthesis-derived electrons....

Evaluation of Time-Temperature Integrators (TTIs) with Microorganism-Entrapped Microbeads Produced Using Homogenization and SPG Membrane Emulsification Techniques.

Science.gov (United States)

Rahman, A T M Mijanur; Lee, Seung Ju; Jung, Seung Won

2015-12-28

A comparative study was conducted to evaluate precision and accuracy in controlling the temperature dependence of encapsulated microbial time-temperature integrators (TTIs) developed using two different emulsification techniques. Weissela cibaria CIFP 009 cells, immobilized within 2% Na-alginate gel microbeads using homogenization (5,000, 7,000, and 10,000 rpm) and Shirasu porous glass (SPG) membrane technologies (10 μm), were applied to microbial TTIs. The prepared micobeads were characterized with respect to their size, size distribution, shape and morphology, entrapment efficiency, and bead production yield. Additionally, fermentation process parameters including growth rate were investigated. The TTI responses (changes in pH and titratable acidity (TA)) were evaluated as a function of temperature (20°C, 25°C, and 30°C). In comparison with conventional methods, SPG membrane technology was able not only to produce highly uniform, small-sized beads with the narrowest size distribution, but also the bead production yield was found to be nearly 3.0 to 4.5 times higher. However, among the TTIs produced using the homogenization technique, poor linearity (R(2)) in terms of TA was observed for the 5,000 and 7,000 rpm treatments. Consequently, microbeads produced by the SPG membrane and by homogenization at 10,000 rpm were selected for adjusting the temperature dependence. The Ea values of TTIs containing 0.5, 1.0, and 1.5 g microbeads, prepared by SPG membrane and conventional methods, were estimated to be 86.0, 83.5, and 76.6 kJ/mol, and 85.5, 73.5, and 62.2 kJ/mol, respectively. Therefore, microbial TTIs developed using SPG membrane technology are much more efficient in controlling temperature dependence.

Generalized chiral membrane dynamics

International Nuclear Information System (INIS)

Cordero, R.; Rojas, E.

2003-01-01

We develop the dynamics of the chiral superconducting membranes (with null current) in an alternative geometrical approach. Besides of this, we show the equivalence of the resulting description with the one known Dirac-Nambu-Goto (DNG) case. Integrability for chiral string model is obtained using a proposed light-cone gauge. In a similar way, domain walls are integrated by means of a simple Ansatz. (Author)

Fabrication and Characterisation of Membrane-Based Gold Electrodes

DEFF Research Database (Denmark)

Bakmand, Tanya; Kwasny, Dorota; Dimaki, Maria

2015-01-01

This work presents a versatile, membrane based electrochemical sensor with thin film electrodes fabricated through Ebeam evaporation directly on porous materials (membranes). Here, the fabrication of the electrodes is described along with possible methods for integration in fluidic systems...

Small RNAs controlling outer membrane porins

DEFF Research Database (Denmark)

Valentin-Hansen, Poul; Johansen, Jesper; Rasmussen, Anders A

2007-01-01

are key regulators of environmental stress. Recent work has revealed an intimate interplay between small RNA regulation of outer membrane proteins and the stress-induced sigmaE-signalling system, which has an essential role in the maintenance of the integrity of the outer membrane.......Gene regulation by small non-coding RNAs has been recognized as an important post-transcriptional regulatory mechanism for several years. In Gram-negative bacteria such as Escherichia coli and Salmonella, these RNAs control stress response and translation of outer membrane proteins and therefore...

On the sub-maximal yield and photo-electric stimulation of chlorophyll a fluorescence in single turnover excitations in plant cells

NARCIS (Netherlands)

Vredenberg, W.J.; Rensen, van J.J.S.; Rodrigues, G.C.

2006-01-01

A set of expressions is derived which quantifies the chlorophyll fluorescence yield in terms of rate constants of primary light reactions of PSII, the fraction of open and semi-open RCs and of the electric field sensed by the RC in the thylakoid membrane. The decay kinetics of the chlorophyll

Oxygen Transport Membranes: A Material Science and Process Engineering Approach

NARCIS (Netherlands)

Chen, Wei

2014-01-01

This thesis describes several fundamental aspects on the membrane-integrated oxy-fuel combustion process and can be divided in two parts: 1) The development and characterization of membrane materials; 2) The design, simulation and evaluation of a coal-fired power plant, coupled with a membrane

Probing lipid membrane electrostatics

Science.gov (United States)

Yang, Yi

The electrostatic properties of lipid bilayer membranes play a significant role in many biological processes. Atomic force microscopy (AFM) is highly sensitive to membrane surface potential in electrolyte solutions. With fully characterized probe tips, AFM can perform quantitative electrostatic analysis of lipid membranes. Electrostatic interactions between Silicon nitride probes and supported zwitterionic dioleoylphosphatidylcholine (DOPC) bilayer with a variable fraction of anionic dioleoylphosphatidylserine (DOPS) were measured by AFM. Classical Gouy-Chapman theory was used to model the membrane electrostatics. The nonlinear Poisson-Boltzmann equation was numerically solved with finite element method to provide the potential distribution around the AFM tips. Theoretical tip-sample electrostatic interactions were calculated with the surface integral of both Maxwell and osmotic stress tensors on tip surface. The measured forces were interpreted with theoretical forces and the resulting surface charge densities of the membrane surfaces were in quantitative agreement with the Gouy-Chapman-Stern model of membrane charge regulation. It was demonstrated that the AFM can quantitatively detect membrane surface potential at a separation of several screening lengths, and that the AFM probe only perturbs the membrane surface potential by external field created by the internai membrane dipole moment. The analysis yields a dipole moment of 1.5 Debye per lipid with a dipole potential of +275 mV for supported DOPC membranes. This new ability to quantitatively measure the membrane dipole density in a noninvasive manner will be useful in identifying the biological effects of the dipole potential. Finally, heterogeneous model membranes were studied with fluid electric force microscopy (FEFM). Electrostatic mapping was demonstrated with 50 nm resolution. The capabilities of quantitative electrostatic measurement and lateral charge density mapping make AFM a unique and powerful

Assembly of Photosynthetic Antenna Protein Complexes from Algae for Development of Nano-biodevice and Its Fuelization

Science.gov (United States)

2013-05-20

bacterial photosynthesis . The structure of the reaction center (RC, the first membrane protein to have its structure determined to high resolution...Introduction] In a bacterial photosynthesis , light-harvesting complex 2 (LH2) and lightharvesting-reaction center complex (LH1-RC) play the key...Artificial Leaf 6CO2 + 6H2O C6H12O6 (Glucose) +6O2 Natural Leaf Photosynthesis and redox proteins are well-organized into thylakoid membrane in natural leaf

Muscle intermediate filaments and their links to membranes and membranous organelles

International Nuclear Information System (INIS)

Capetanaki, Yassemi; Bloch, Robert J.; Kouloumenta, Asimina; Mavroidis, Manolis; Psarras, Stelios

2007-01-01

Intermediate filaments (IFs) play a key role in the integration of structure and function of striated muscle, primarily by mediating mechanochemical links between the contractile apparatus and mitochondria, myonuclei, the sarcolemma and potentially the vesicle trafficking apparatus. Linkage of all these membranous structures to the contractile apparatus, mainly through the Z-disks, supports the integration and coordination of growth and energy demands of the working myocyte, not only with force transmission, but also with de novo gene expression, energy production and efficient protein and lipid trafficking and targeting. Desmin, the most abundant and intensively studied muscle intermediate filament protein, is linked to proper costamere organization, myoblast and stem cell fusion and differentiation, nuclear shape and positioning, as well as mitochondrial shape, structure, positioning and function. Similar links have been established for lysosomes and lysosome-related organelles, consistent with the presence of widespread links between IFs and membranous structures and the regulation of their fusion, morphology and stabilization necessary for cell survival

Structure of an E. coli integral membrane sulfurtransferase and its structural transition upon SCN− binding defined by EPR-based hybrid method

Science.gov (United States)

Ling, Shenglong; Wang, Wei; Yu, Lu; Peng, Junhui; Cai, Xiaoying; Xiong, Ying; Hayati, Zahra; Zhang, Longhua; Zhang, Zhiyong; Song, Likai; Tian, Changlin

2016-01-01

Electron paramagnetic resonance (EPR)-based hybrid experimental and computational approaches were applied to determine the structure of a full-length E. coli integral membrane sulfurtransferase, dimeric YgaP, and its structural and dynamic changes upon ligand binding. The solution NMR structures of the YgaP transmembrane domain (TMD) and cytosolic catalytic rhodanese domain were reported recently, but the tertiary fold of full-length YgaP was not yet available. Here, systematic site-specific EPR analysis defined a helix-loop-helix secondary structure of the YagP-TMD monomers using mobility, accessibility and membrane immersion measurements. The tertiary folds of dimeric YgaP-TMD and full-length YgaP in detergent micelles were determined through inter- and intra-monomer distance mapping and rigid-body computation. Further EPR analysis demonstrated the tight packing of the two YgaP second transmembrane helices upon binding of the catalytic product SCN−, which provides insight into the thiocyanate exportation mechanism of YgaP in the E. coli membrane. PMID:26817826

A micromachined membrane-based active probe for biomolecular mechanics measurement

Science.gov (United States)

Torun, H.; Sutanto, J.; Sarangapani, K. K.; Joseph, P.; Degertekin, F. L.; Zhu, C.

2007-04-01

A novel micromachined, membrane-based probe has been developed and fabricated as assays to enable parallel measurements. Each probe in the array can be individually actuated, and the membrane displacement can be measured with high resolution using an integrated diffraction-based optical interferometer. To illustrate its application in single-molecule mechanics experiments, this membrane probe was used to measure unbinding forces between L-selectin reconstituted in a polymer-cushioned lipid bilayer on the probe membrane and an antibody adsorbed on an atomic force microscope cantilever. Piconewton range forces between single pairs of interacting molecules were measured from the cantilever bending while using the membrane probe as an actuator. The integrated diffraction-based optical interferometer of the probe was demonstrated to have floor for frequencies as low as 3 Hz with a differential readout scheme. With soft probe membranes, this low noise level would be suitable for direct force measurements without the need for a cantilever. Furthermore, the probe membranes were shown to have 0.5 µm actuation range with a flat response up to 100 kHz, enabling measurements at fast speeds.

Composite membranes and methods for making same

Science.gov (United States)

Routkevitch, Dmitri; Polyakov, Oleg G

2012-07-03

Composite membranes that are adapted for separation, purification, filtration, analysis, reaction and sensing. The composite membranes can include a porous support structure having elongate pore channels extending through the support structure. The composite membrane also includes an active layer comprising an active layer material, where the active layer material is completely disposed within the pore channels between the surfaces of the support structure. The active layer is intimately integrated within the support structure, thus enabling great robustness, reliability, resistance to mechanical stress and thermal cycling, and high selectivity. Methods for the fabrication of composite membranes are also provided.

Integrated Microfluidic Membrane Transistor Utilizing Chemical Information for On-Chip Flow Control

Science.gov (United States)

Frank, Philipp; Schreiter, Joerg; Haefner, Sebastian; Paschew, Georgi; Voigt, Andreas; Richter, Andreas

2016-01-01

Microfluidics is a great enabling technology for biology, biotechnology, chemistry and general life sciences. Despite many promising predictions of its progress, microfluidics has not reached its full potential yet. To unleash this potential, we propose the use of intrinsically active hydrogels, which work as sensors and actuators at the same time, in microfluidic channel networks. These materials transfer a chemical input signal such as a substance concentration into a mechanical output. This way chemical information is processed and analyzed on the spot without the need for an external control unit. Inspired by the development electronics, our approach focuses on the development of single transistor-like components, which have the potential to be used in an integrated circuit technology. Here, we present membrane isolated chemical volume phase transition transistor (MIS-CVPT). The device is characterized in terms of the flow rate from source to drain, depending on the chemical concentration in the control channel, the source-drain pressure drop and the operating temperature. PMID:27571209

Integrated Microfluidic Membrane Transistor Utilizing Chemical Information for On-Chip Flow Control.

Science.gov (United States)

Frank, Philipp; Schreiter, Joerg; Haefner, Sebastian; Paschew, Georgi; Voigt, Andreas; Richter, Andreas

2016-01-01

Microfluidics is a great enabling technology for biology, biotechnology, chemistry and general life sciences. Despite many promising predictions of its progress, microfluidics has not reached its full potential yet. To unleash this potential, we propose the use of intrinsically active hydrogels, which work as sensors and actuators at the same time, in microfluidic channel networks. These materials transfer a chemical input signal such as a substance concentration into a mechanical output. This way chemical information is processed and analyzed on the spot without the need for an external control unit. Inspired by the development electronics, our approach focuses on the development of single transistor-like components, which have the potential to be used in an integrated circuit technology. Here, we present membrane isolated chemical volume phase transition transistor (MIS-CVPT). The device is characterized in terms of the flow rate from source to drain, depending on the chemical concentration in the control channel, the source-drain pressure drop and the operating temperature.

On the mechanism of sulfite activation of chloroplast thylakoid ATPase and the relation of ADP tightly bound at a catalytic site to the binding change mechanism

International Nuclear Information System (INIS)

Du, Z.; Boyer, P.D.

1990-01-01

Washed chloroplast thylakoid membranes upon exposure to [ 3 H]ADP retain in tightly bound [ 3 H]ADP on a catalytic site of the ATP synthase. The presence of sufficient endogenous or added Mg 2+ results in an enzyme with essentially no ATPase activity. Sulfite activates the ATPase, and many molecules of ATP per synthase can be hydrolyzed before most of the bound [ 3 H]ADP is released, a result interpreted as indicating that the ADP is not bound at a site participating in catalysis by the sulfite-activated enzyme. The authors present evidence that this is not the case. The Mg 2+ - and ADP-inhibited enzyme when exposed to MgATP and 20-100 mM sulfite shows a lag of about 1 min at 22 degree C and of about 15 s at 37 degree C before reaching the same steady-state rate as attained with light-activated ATPase that has not been inhibited by Mg 2+ and ADP. The lag is not eliminated if the enzyme is exposed to sulfite prior to MgATP addition, indicating that ATPase turnover is necessary for the activation. The release of most of the bound [ 3 H]ADP parallels the onset of ATPase activity, although some [ 3 H]ADP is not released even with prolonged catalytic turnover and may be on poorly active or inactive enzyme or at noncatalytic sites. The results are consistent with most of the tightly bound [ 3 H]ADP being at a catalytic site and being replaced as this Mg 2+ - and ADP-inhibited site regains equivalent participation with other catalytic sites on the activated enzyme. The sulfite activation can be explained by sulfite combination at a P i binding site of the enzyme-ADP-Mg 2+ complex to give a form more readily activated by ATP binding at an alternative site

Lipid antioxidant and galactolipid remodeling under temperature stress in tomato plants

Directory of Open Access Journals (Sweden)

Livia eSpicher

2016-02-01

Full Text Available Increased temperatures are a major scenario in climate change and present a threat to plant growth and agriculture. Plant growth depends on photosynthesis. To function optimally the photosynthetic machinery at the thylakoid membrane in chloroplasts continuously adapts to changing conditions. Here, we set out to discover the most important changes arising at the lipid level under high temperature (38°C in comparison to mild (20°C and moderately cold temperature (10°C using a non-targeted lipidomics approach. To our knowledge, no comparable experiment at the level of the whole membrane system has been documented. Here, 791 molecular species were detected by mass spectrometry and ranged from membrane lipids, prenylquinones (tocopherols, phylloquinone, plastoquinone, plastochromanol, carotenoids (β-carotene, xanthophylls to numerous unidentified compounds. At high temperatures, the most striking changes were observed for the prenylquinones (α-tocopherol and plastoquinone/-ol and the degree of saturation of fatty acids in galactolipids and phosphatidyl ethanolamine. Photosynthetic efficiency at high temperature was not affected but at moderately cold temperature mild photoinhibition occurred. The results indicate that the thylakoid membrane is remodeled with regard to fatty acid saturation in galactolipids and lipid antioxidant concentrations under high temperature stress. The data strongly suggest that massively increased concentrations of α-tocopherol and plastoquinone are important for protection against high temperature stress and proper function of the photosynthetic apparatus.

The vascular basement membrane in the healthy and pathological brain.

Science.gov (United States)

Thomsen, Maj S; Routhe, Lisa J; Moos, Torben

2017-10-01

The vascular basement membrane contributes to the integrity of the blood-brain barrier (BBB), which is formed by brain capillary endothelial cells (BCECs). The BCECs receive support from pericytes embedded in the vascular basement membrane and from astrocyte endfeet. The vascular basement membrane forms a three-dimensional protein network predominantly composed of laminin, collagen IV, nidogen, and heparan sulfate proteoglycans that mutually support interactions between BCECs, pericytes, and astrocytes. Major changes in the molecular composition of the vascular basement membrane are observed in acute and chronic neuropathological settings. In the present review, we cover the significance of the vascular basement membrane in the healthy and pathological brain. In stroke, loss of BBB integrity is accompanied by upregulation of proteolytic enzymes and degradation of vascular basement membrane proteins. There is yet no causal relationship between expression or activity of matrix proteases and the degradation of vascular matrix proteins in vivo. In Alzheimer's disease, changes in the vascular basement membrane include accumulation of Aβ, composite changes, and thickening. The physical properties of the vascular basement membrane carry the potential of obstructing drug delivery to the brain, e.g. thickening of the basement membrane can affect drug delivery to the brain, especially the delivery of nanoparticles.

Outer membrane protein functions as integrator of protein import and DNA inheritance in mitochondria

Science.gov (United States)

Käser, Sandro; Oeljeklaus, Silke; Týč, Jiří; Vaughan, Sue; Warscheid, Bettina; Schneider, André

2016-01-01

Trypanosomatids are one of the earliest diverging eukaryotes that have fully functional mitochondria. pATOM36 is a trypanosomatid-specific essential mitochondrial outer membrane protein that has been implicated in protein import. Changes in the mitochondrial proteome induced by ablation of pATOM36 and in vitro assays show that pATOM36 is required for the assembly of the archaic translocase of the outer membrane (ATOM), the functional analog of the TOM complex in other organisms. Reciprocal pull-down experiments and immunofluorescence analyses demonstrate that a fraction of pATOM36 interacts and colocalizes with TAC65, a previously uncharacterized essential component of the tripartite attachment complex (TAC). The TAC links the single-unit mitochondrial genome to the basal body of the flagellum and mediates the segregation of the replicated mitochondrial genomes. RNAi experiments show that pATOM36, in line with its dual localization, is not only essential for ATOM complex assembly but also for segregation of the replicated mitochondrial genomes. However, the two functions are distinct, as a truncated version of pATOM36 lacking the 75 C-terminal amino acids can rescue kinetoplast DNA missegregation but not the lack of ATOM complex assembly. Thus, pATOM36 has a dual function and integrates mitochondrial protein import with mitochondrial DNA inheritance. PMID:27436903

The DnaJ-like zinc finger domain protein PSA2 affects light acclimation and chloroplast development in Arabidopsis thaliana

Directory of Open Access Journals (Sweden)

Yan-Wen eWang

2016-03-01

Full Text Available The biosynthesis of chlorophylls and carotenoids and the assembly of thylakoid membranes are critical for the photoautotrophic growth of plants. Different factors are involved in these two processes. In recent years, members of the DnaJ-like zinc finger domain proteins have been found to take part in the biogenesis and/or the maintenance of plastids. One member of this family of proteins, PSA2, was recently found to localize to the thylakoid lumen and regulate the accumulation of photosystem I. In this study, we report that the silencing of PSA2 in Arabidopsis thaliana resulted in variegated leaves and retarded growth. Although both chlorophylls and total carotenoids decreased in the psa2 mutant, violaxanthin and zeaxanthin accumulated in the mutant seedlings grown under growth condition. Lower levels of non-photochemical quenching and electron transport rate were also found in the psa2 mutant seedlings under growth condition compared with those of the wild-type plants, indicating an impaired capability to acclimate to normal light irradiance when PSA2 was silenced. Moreover, we also observed an abnormal assembly of grana thylakoids and poorly developed stroma thylakoids in psa2 chloroplasts. Taken together, our results demonstrate that PSA2 is a member of the DnaJ-like zinc finger domain protein family that affects light acclimation and chloroplast development.

Profiling the outer membrane proteome during growth and development of the social bacterium Myxococcus xanthus by selective biotinylation and analyses of outer membrane vesicles.

Science.gov (United States)

Kahnt, Jörg; Aguiluz, Kryssia; Koch, Jürgen; Treuner-Lange, Anke; Konovalova, Anna; Huntley, Stuart; Hoppert, Michael; Søgaard-Andersen, Lotte; Hedderich, Reiner

2010-10-01

Social behavior in the bacterium Myxococcus xanthus relies on contact-dependent activities involving cell-cell and cell-substratum interactions. To identify outer membrane proteins that have a role in these activities, we profiled the outer membrane proteome of growing and starving cells using two strategies. First, outer membrane proteins were enriched by biotinylation of intact cells using the reagent NHS (N-hydroxysuccinimide)-PEO(12) (polyethylene oxide)-biotin with subsequent membrane solubilization and affinity chromatography. Second, the proteome of outer membrane vesicles (OMV) was determined. Comparisons of detected proteins show that these methods have different detection profiles and together provide a comprehensive view of the outer membrane proteome. From 362 proteins identified, 274 (76%) were cell envelope proteins including 64 integral outer membrane proteins and 85 lipoproteins. The majority of these proteins were of unknown function. Among integral outer membrane proteins with homologues of known function, TonB-dependent transporters comprise the largest group. Our data suggest novel functions for these transporters. Among lipoproteins with homologues of known function, proteins with hydrolytic functions comprise the largest group. The luminal load of OMV was enriched for proteins with hydrolytic functions. Our data suggest that OMV have functions in predation and possibly in transfer of intercellular signaling molecules between cells.

High speed municipal sewage treatment in microbial fuel cell integrated with anaerobic membrane filtration system.

Science.gov (United States)

Lee, Y; Oa, S W

2014-01-01

A cylindrical two chambered microbial fuel cell (MFC) integrated with an anaerobic membrane filter was designed and constructed to evaluate bioelectricity generation and removal efficiency of organic substrate (glucose or domestic wastewater) depending on organic loading rates (OLRs). The MFC was continuously operated with OLRs 3.75, 5.0, 6.25, and 9.38 kg chemical oxygen demand (COD)/(m(3)·d) using glucose as a substrate, and the cathode chamber was maintained at 5-7 mg/L of dissolved oxygen. The optimal OLR was found to be 6.25 kgCOD/(m(3)·d) (hydraulic retention time (HRT) 1.9 h), and the corresponding voltage and power density averaged during the operation were 0.15 V and 13.6 mW/m(3). With OLR 6.25 kgCOD/(m(3)·d) using domestic wastewater as a substrate, the voltage and power reached to 0.13 V and 91 mW/m(3) in the air cathode system. Even though a relatively short HRT of 1.9 h was applied, stable effluent could be obtained by the membrane filtration system and the following air purging. In addition, the short HRT would provide economic benefit in terms of reduction of construction and operating costs compared with a conventional aerobic treatment process.

Microporous Silica Based Membranes for Desalination

Directory of Open Access Journals (Sweden)

João C. Diniz da Costa

2012-09-01

Full Text Available This review provides a global overview of microporous silica based membranes for desalination via pervaporation with a focus on membrane synthesis and processing, transport mechanisms and current state of the art membrane performance. Most importantly, the recent development and novel concepts for improving the hydro-stability and separating performance of silica membranes for desalination are critically examined. Research into silica based membranes for desalination has focussed on three primary methods for improving the hydro-stability. These include incorporating carbon templates into the microporous silica both as surfactants and hybrid organic-inorganic structures and incorporation of metal oxide nanoparticles into the silica matrix. The literature examined identified that only metal oxide silica membranes have demonstrated high salt rejections under a variety of feed concentrations, reasonable fluxes and unaltered performance over long-term operation. As this is an embryonic field of research several target areas for researchers were discussed including further improvement of the membrane materials, but also regarding the necessity of integrating waste or solar heat sources into the final process design to ensure cost competitiveness with conventional reverse osmosis processes.

Thermo-optically induced reorganizations in the main light harvesting antenna of plants. II

DEFF Research Database (Denmark)

Holm, Jens Kai; Varkonyi, Zsuzsanna; Kovacs, Laszlo

2005-01-01

We have investigated the circular dichroism spectral transients associated with the light-induced reversible reorganizations in chirally organized macrodomains of pea thylakoid membranes and loosely stacked lamellar aggregates of the main chlorophyll a/b light harvesting complexes (LHCII) isolated...... from the same membranes. These reorganizations have earlier been assigned to originate from a thermo-optic effect. According to the thermo-optic mechanism, fast local thermal transients due to dissipation of the excess excitation energy induce elementary structural changes in the close vicinity...

Infrared emission of a freestanding plasmonic membrane

Science.gov (United States)

Monshat, Hosein; Liu, Longju; McClelland, John; Biswas, Rana; Lu, Meng

2018-01-01

This paper reports a free-standing plasmonic membrane as a thermal emitter in the near- and mid-infrared regions. The plasmonic membrane consists of an ultrathin gold film perforated with a two-dimensional array of holes. The device was fabricated using an imprint and transfer process and fixed on a low-emissivity metal grid. The thermal radiation characteristics of the plasmonic membrane can be engineered by controlling the array period and the thickness of the gold membrane. Plasmonic membranes with two different periods were designed using electromagnetic simulation and then characterized for their transmission and infrared radiation properties. The free-standing membranes exhibit extraordinary optical transmissions with the resonant transmission coefficient as high as 76.8%. After integration with a customized heater, the membranes demonstrate narrowband thermal emission in the wavelength range of 2.5 μm to 5.5 μm. The emission signatures, including peak emission wavelength and bandwidth, are associated with the membrane geometry. The ultrathin membrane infrared emitter can be adopted in applications, such as chemical analysis and thermal imaging.

Biogenesis of light harvesting proteins.

Science.gov (United States)

Dall'Osto, Luca; Bressan, Mauro; Bassi, Roberto

2015-09-01

The LHC family includes nuclear-encoded, integral thylakoid membrane proteins, most of which coordinate chlorophyll and xanthophyll chromophores. By assembling with the core complexes of both photosystems, LHCs form a flexible peripheral moiety for enhancing light-harvesting cross-section, regulating its efficiency and providing protection against photo-oxidative stress. Upon its first appearance, LHC proteins underwent evolutionary diversification into a large protein family with a complex genetic redundancy. Such differentiation appears as a crucial event in the adaptation of photosynthetic organisms to changing environmental conditions and land colonization. The structure of photosystems, including nuclear- and chloroplast-encoded subunits, presented the cell with a number of challenges for the control of the light harvesting function. Indeed, LHC-encoding messages are translated in the cytosol, and pre-proteins imported into the chloroplast, processed to their mature size and targeted to the thylakoids where are assembled with chromophores. Thus, a tight coordination between nuclear and plastid gene expression, in response to environmental stimuli, is required to adjust LHC composition during photoacclimation. In recent years, remarkable progress has been achieved in elucidating structure, function and regulatory pathways involving LHCs; however, a number of molecular details still await elucidation. In this review, we will provide an overview on the current knowledge on LHC biogenesis, ranging from organization of pigment-protein complexes to the modulation of gene expression, import and targeting to the photosynthetic membranes, and regulation of LHC assembly and turnover. Genes controlling these events are potential candidate for biotechnological applications aimed at optimizing light use efficiency of photosynthetic organisms. This article is part of a Special Issue entitled: Chloroplast biogenesis. Copyright © 2015 Elsevier B.V. All rights reserved.

Photosynthetic properties of erect leaf maize inbred lines as the efficient photo-model in breeding and seed production

Directory of Open Access Journals (Sweden)

Radenović Čedomir N.

2003-01-01

Full Text Available The initial idea of this study was a hypothesis that erect leaf maize inbred lines were characterized by properties of an efficient photo-model and that as such were very desirable in increasing the number of plants per area unit (plant density in the process of contemporary selection and seed production. The application of a non-invasive bioluminescence-photosynthetic method, suitable for the efficiency estimation of the photo-model, verified the hypothesis. Obtained photosynthetic properties of observed erect leaf maize inbred lines were based on the effects and characteristics of thermal processes of delayed chlorophyll fluorescence occurring in their thylakoid membranes. The temperature dependence of the delayed chlorophyll fluorescence intensity phase transitions (critical temperatures in the thylakoid membranes and activation energy are the principal parameters of the thermal processes. Based on obtained photosynthetic properties it is possible to select erect leaf maize inbred lines that are resistant and tolerant to high and very high temperatures, as well as, to drought. They could be good and efficient photo-models wherewith.

Osmotic versus conventional membrane bioreactors integrated with reverse osmosis for water reuse: Biological stability, membrane fouling, and contaminant removal.

Science.gov (United States)

Luo, Wenhai; Phan, Hop V; Xie, Ming; Hai, Faisal I; Price, William E; Elimelech, Menachem; Nghiem, Long D

2017-02-01

This study systematically compares the performance of osmotic membrane bioreactor - reverse osmosis (OMBR-RO) and conventional membrane bioreactor - reverse osmosis (MBR-RO) for advanced wastewater treatment and water reuse. Both systems achieved effective removal of bulk organic matter and nutrients, and almost complete removal of all 31 trace organic contaminants investigated. They both could produce high quality water suitable for recycling applications. During OMBR-RO operation, salinity build-up in the bioreactor reduced the water flux and negatively impacted the system biological treatment by altering biomass characteristics and microbial community structure. In addition, the elevated salinity also increased soluble microbial products and extracellular polymeric substances in the mixed liquor, which induced fouling of the forward osmosis (FO) membrane. Nevertheless, microbial analysis indicated that salinity stress resulted in the development of halotolerant bacteria, consequently sustaining biodegradation in the OMBR system. By contrast, biological performance was relatively stable throughout conventional MBR-RO operation. Compared to conventional MBR-RO, the FO process effectively prevented foulants from permeating into the draw solution, thereby significantly reducing fouling of the downstream RO membrane in OMBR-RO operation. Accumulation of organic matter, including humic- and protein-like substances, as well as inorganic salts in the MBR effluent resulted in severe RO membrane fouling in conventional MBR-RO operation. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

High-throughput Cloning and Expression of Integral Membrane Proteins in Escherichia coli

Science.gov (United States)

Bruni, Renato

2014-01-01

Recently, several structural genomics centers have been established and a remarkable number of three-dimensional structures of soluble proteins have been solved. For membrane proteins, the number of structures solved has been significantly trailing those for their soluble counterparts, not least because over-expression and purification of membrane proteins is a much more arduous process. By using high throughput technologies, a large number of membrane protein targets can be screened simultaneously and a greater number of expression and purification conditions can be employed, leading to a higher probability of successfully determining the structure of membrane proteins. This unit describes the cloning, expression and screening of membrane proteins using high throughput methodologies developed in our laboratory. Basic Protocol 1 deals with the cloning of inserts into expression vectors by ligation-independent cloning. Basic Protocol 2 describes the expression and purification of the target proteins on a miniscale. Lastly, for the targets that express at the miniscale, basic protocols 3 and 4 outline the methods employed for the expression and purification of targets at the midi-scale, as well as a procedure for detergent screening and identification of detergent(s) in which the target protein is stable. PMID:24510647

Ophiobolin A from Bipolaris oryzae Perturbs Motility and Membrane Integrities of Porcine Sperm and Induces Cell Death on Mammalian Somatic Cell Lines

Directory of Open Access Journals (Sweden)

Ottó Bencsik

2014-09-01

Full Text Available Bipolaris oryzae is a phytopathogenic fungus causing a brown spot disease in rice, and produces substance that strongly perturbs motility and membrane integrities of boar spermatozoa. The substance was isolated from the liquid culture of the fungal strain using extraction and a multi-step semi-preparative HPLC procedures. Based on the results of mass spectrometric and 2D NMR techniques, the bioactive molecule was identified as ophiobolin A, a previously described sesterterpene-type compound. The purified ophiobolin A exhibited strong motility inhibition and viability reduction on boar spermatozoa. Furthermore, it damaged the sperm mitochondria significantly at sublethal concentration by the dissipation of transmembrane potential in the mitochondrial inner membrane, while the plasma membrane permeability barrier remained intact. The study demonstrated that the cytotoxicity of ophiobolin A toward somatic cell lines is higher by 1–2 orders of magnitude compared to other mitochondriotoxic mycotoxins, and towards sperm cells unique by replacing the progressive motility by shivering tail beating at low exposure concentration.

Mitofilin complexes: conserved organizers of mitochondrial membrane architecture.

Science.gov (United States)

Zerbes, Ralf M; van der Klei, Ida J; Veenhuis, Marten; Pfanner, Nikolaus; van der Laan, Martin; Bohnert, Maria

2012-11-01

Mitofilin proteins are crucial organizers of mitochondrial architecture. They are located in the inner mitochondrial membrane and interact with several protein complexes of the outer membrane, thereby generating contact sites between the two membrane systems of mitochondria. Within the inner membrane, mitofilins are part of hetero-oligomeric protein complexes that have been termed the mitochondrial inner membrane organizing system (MINOS). MINOS integrity is required for the maintenance of the characteristic morphology of the inner mitochondrial membrane, with an inner boundary region closely apposed to the outer membrane and cristae membranes, which form large tubular invaginations that protrude into the mitochondrial matrix and harbor the enzyme complexes of the oxidative phosphorylation machinery. MINOS deficiency comes along with a loss of crista junction structures and the detachment of cristae from the inner boundary membrane. MINOS has been conserved in evolution from unicellular eukaryotes to humans, where alterations of MINOS subunits are associated with multiple pathological conditions.

Performance modeling of direct contact membrane distillation (DCMD) seawater desalination process using a commercial composite membrane

KAUST Repository

Lee, Junggil

2015-01-10

This paper presents the development of a rigorous theoretical model to predict the transmembrane flux of a flat sheet hydrophobic composite membrane, comprising both an active layer of polytetrafluoroethylene and a scrim-backing support layer of polypropylene, in the direct contact membrane distillation (DCMD) process. An integrated model includes the mass, momentum, species and energy balances for both retentate and permeate flows, coupled with the mass transfer of water vapor through the composite membrane and the heat transfer across the membrane and through the boundary layers adjacent to the membrane surfaces. Experimental results and model predictions for permeate flux and performance ratio are compared and shown to be in good agreement. The permeate flux through the composite layer can be ignored in the consideration of mass transfer pathways at the composite membrane. The effect of the surface porosity and the thickness of active and support layers on the process performance of composite membrane has also been studied. Among these parameters, surface porosity is identified to be the main factor significantly influencing the permeate flux and performance ratio, while the relative influence of the surface porosity on the performance ratio is less than that on flux.

Modeling a Membrane: Using Engineering Design to Simulate Cell Transport Processes

Science.gov (United States)

Mason, Kevin; Evans, Brian

2017-01-01

The "plasma membrane," which controls what comes in and goes out of a cell, is integral to maintaining homeostasis. Cell transport of small molecules across the cell membrane happens in several different ways. Some small, nonpolar molecules cross the plasma membrane along the concentration gradient directly through the "phospholipid…

Integration of membrane distillation into traditional salt farming method: Process development and modelling

Science.gov (United States)

Hizam, S.; Bilad, M. R.; Putra, Z. A.

2017-10-01

Farmers still practice the traditional salt farming in many regions, particularly in Indonesia. This archaic method not only produces low yield and poor salt quality, it is also laborious. Furthermore, the farming locations typically have poor access to fresh water and are far away from electricity grid, which restrict upgrade to a more advanced technology for salt production. This paper proposes a new concept of salt harvesting method that improves the salt yield and at the same time facilitates recovery of fresh water from seawater. The new concept integrates solar powered membrane distillation (MD) and photovoltaic cells to drive the pumping. We performed basic solar still experiments to quantify the heat flux received by a pond. The data were used as insight for designing the proposed concept, particularly on operational strategy and the most effective way to integrate MD. After the conceptual design had been developed, we formulated mass and energy balance to estimate the performance of the proposed concept. Based on our data and design, it is expected that the system would improve the yield and quality of the salt production, maximizing fresh water harvesting, and eventually provides economical gain for salt farmers hence improving their quality of life. The key performance can only be measured via experiment using gain output ratio as performance indicator, which will be done in a future study.

Pressure effects on lipids and bio-membrane assemblies

Directory of Open Access Journals (Sweden)

Nicholas J. Brooks

2014-11-01

Full Text Available Membranes are amongst the most important biological structures; they maintain the fundamental integrity of cells, compartmentalize regions within them and play an active role in a wide range of cellular processes. Pressure can play a key role in probing the structure and dynamics of membrane assemblies, and is also critical to the biology and adaptation of deep-sea organisms. This article presents an overview of the effect of pressure on the mesostructure of lipid membranes, bilayer organization and lipid–protein assemblies. It also summarizes recent developments in high-pressure structural instrumentation suitable for experiments on membranes.

Proton Conductivity and Operational Features Of PBI-Based Membranes

DEFF Research Database (Denmark)

Qingfeng, Li; Jensen, Jens Oluf; Precht Noyé, Pernille

2005-01-01

As an approach to high temperature operation of PEMFCs, acid-doped PBI membranes are under active development. The membrane exhibits high proton conductivity under low water contents at temperatures up to 200°C. Mechanisms of proton conduction for the membranes have been proposed. Based on the me...... on the membranes fuel cell tests have been demonstrated. Operating features of the PBI cell include no humidification, high CO tolerance, better heat utilization and possible integration with fuel processing units. Issues for further development are also discussed....

Purification and partial characterization of analogous 26-kDa rat submandibular and parotid gland integral membrane phosphoproteins that may have a role in exocytosis.

Science.gov (United States)

Quissell, D O; Deisher, L M

1992-04-01

Rat submandibular and parotid gland exocytosis is primarily controlled by beta-adrenergic receptor stimulation. Although its precise role in the regulation of salivary gland exocytosis is not fully understood, protein phosphorylation, mediated by the activation of cAMP-dependent protein kinase, may be directly involved. Previous studies suggest that analogous 26-kDa integral membrane phosphoproteins may play a direct role in regulating exocytosis. Studies were here undertaken to purify and partially characterize both phosphoproteins. After endogenous phosphorylation with 32P, subcellular fraction and solubilization of the microsomal fraction in n-octyl beta-glucopyranoside, the 26-kDa integral membrane phosphoproteins were purified by high performance liquid chromatography (HPLC), followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and electroelution of the proteins. Amino acid analysis indicated a significant number of serine amino acids: N-terminal sequence data demonstrated a high level of homology; and trypsin digestion followed by reversed-phase HPLC indicated the possibility of multiple phosphorylation sites.

Integration of biological method and membrane technology in treating palm oil mill effluent

Institute of Scientific and Technical Information of China (English)

ZHANG Yejian; YAN Li; QIAO Xiangli; CHI Lina; NIU Xiangjun; MEI Zhijian; ZHANG Zhenjia

2008-01-01

Palm oil industry is the most important agro-industry in Malaysia, but its by-product-palm oil mill effluent (POME), posed a great threat to water environment. In the past decades, several treatment and disposal methods have been proposed and investigated to solve this problem. A two-stage pilot-scale plant was designed and constructed for POME treatment. Anaerobic digestion and aerobic biodegradation constituted the first biological stage, while ultrafiltration (UF) and reverse osmosis (RO) membrane units were combined as the second membrane separation stage. In the anaerobic expanded granular sludge bed (EGSB) reactor, about 43% organic matter in POME was converted into biogas, and COD reduction efficiency reached 93% and 22% in EGSB and the following aerobic reactor, respectively. With the treatment in the first biological stage, suspended solids and oil also decreased to a low degree. All these alleviated the membrane fouling and prolonged the membrane life. In the membrane process unit, almost all the suspended solids were captured by UF membranes, while RO membrane excluded most of the dissolved solids or inorganic salts from RO permeate. After the whole treatment processes, organic matter in POME expressed by BOD and COD was removed almost thoroughly. Suspended solids and color were not detectable in RO permeate any more, and mineral elements only existed in trace amount (except for K and Na). The high-quality effluent was crystal clear and could be used as the boiler feed water.

Counter-current membrane reactor for WGS process: Membrane design

Energy Technology Data Exchange (ETDEWEB)

Piemonte, Vincenzo; Favetta, Barbara [Department of Chemical Engineering Materials and Environment, University of Rome ' ' La Sapienza' ' , via Eudossiana 18, 00184 Rome (Italy); De Falco, Marcello [Faculty of Engineering, University Campus Bio-Medico of Rome, via Alvaro del Portillo 21, 00128 Rome (Italy); Basile, Angelo [CNR-ITM, c/o University of Calabria, Via Pietro Bucci, Cubo 17/C, 87030 Rende (CS) (Italy)

2010-11-15

Water gas shift (WGS) is a thermodynamically limited reaction which has to operate at low temperatures, reducing kinetics rate and increasing the amount of catalyst required to reach valuable CO conversions. It has been widely demonstrated that the integration of hydrogen selective membranes is a promising way to enhance WGS reactors performance: a Pd-based MR operated successfully overcoming the thermodynamic constraints of a traditional reactor thanks to the removal of hydrogen from reaction environment. In the first part of a MR, the H{sub 2} partial pressure starts from a minimum value since the reaction has not started. As a consequence, if the carrier gas in the permeation zone is sent in counter-current, which is the most efficient configuration, in the first reactor section the H{sub 2} partial pressure in reaction zone is low while in the permeation zone is high, potentially implying back permeation. This means a bad utilization of the first part of the membrane area and thus, a worsening of the MR performance with lower H{sub 2} recovery and lower CO conversion with respect to the case in which the whole selective surface is properly used. To avoid this problem different MR configurations were evaluated by a 1-D pseudo-homogeneous model, validated with WGS industrial data reported in scientific literature. It was demonstrated that the permeated H{sub 2} flow rate per membrane surface, i.e. the membrane flux, strongly improves if selective membrane is placed only in the second part of the reactor: in fact, if the membrane is placed at L{sub m}/L{sub tot} = 0.5, the membrane flux is 0.2 kmol/(m{sup 2}h) about, if it is placed along all reactor tube (L{sub m}/L{sub tot} = 1), flux is 0.05 kmol/(m{sup 2}h). The effect of the L/D reactor ratio and of the reactor wall temperature on the CO conversion were also assessed. (author)

Plasma membrane and salinity tolerance of barley plants

International Nuclear Information System (INIS)

Al-Rahmani, F. H.; Al-Mashhadani, M. S.; Al-Delemee, N. H.

1997-01-01

Barley cultivar, California Mario ut, was grown in a nutrient solution containing increasing Nacl concentrations up to 250 mm. The effect of Nacl on growth, mineral compost ion ant integrity of the plasma membrane was studied. Growth of the shoot'and root was stimulated or little affected by 10 and 20 ml Nacl. Further increase in Nacl concentrations depressed the growth. The depression was conspicuous between 100 and 250 mm Nacl. Increasing Nacl concentration decreased potassium content in the shoots and roots and led to steep increase in sodium accumulation. The integrity of the plasma membrane was measured in term of potassium leakage from the root tips. Rapid leakage of potassium was obtained at Nacl concentrations ranging from 100 to 250 mm. At the same concentrations of Nacl, adenosine triphosphatase activity in the root tips was increased. Results indicate that the plasma membrane of root cells was damaged by the increased levels of salinity. It was concluded that the plasma membrane of root cells is the primary site of salinity toxicity. (authors). 40 refs., 5 tabs. 3 figs

A chloroplast thylakoid lumen protein is required for proper photosynthetic acclimation of plants under fluctuating light environments.

Science.gov (United States)

Liu, Jun; Last, Robert L

2017-09-19

Despite our increasingly sophisticated understanding of mechanisms ensuring efficient photosynthesis under laboratory-controlled light conditions, less is known about the regulation of photosynthesis under fluctuating light. This is important because-in nature-photosynthetic organisms experience rapid and extreme changes in sunlight, potentially causing deleterious effects on photosynthetic efficiency and productivity. Here we report that the chloroplast thylakoid lumenal protein MAINTENANCE OF PHOTOSYSTEM II UNDER HIGH LIGHT 2 (MPH2; encoded by At4g02530 ) is required for growth acclimation of Arabidopsis thaliana plants under controlled photoinhibitory light and fluctuating light environments. Evidence is presented that mph2 mutant light stress susceptibility results from a defect in photosystem II (PSII) repair, and our results are consistent with the hypothesis that MPH2 is involved in disassembling monomeric complexes during regeneration of dimeric functional PSII supercomplexes. Moreover, mph2 -and previously characterized PSII repair-defective mutants-exhibited reduced growth under fluctuating light conditions, while PSII photoprotection-impaired mutants did not. These findings suggest that repair is not only required for PSII maintenance under static high-irradiance light conditions but is also a regulatory mechanism facilitating photosynthetic adaptation under fluctuating light environments. This work has implications for improvement of agricultural plant productivity through engineering PSII repair.

Step-by-step seeding procedure for preparing HKUST-1 membrane on porous α-alumina support.

Science.gov (United States)

Nan, Jiangpu; Dong, Xueliang; Wang, Wenjin; Jin, Wanqin; Xu, Nanping

2011-04-19

Metal-organic framework (MOF) membranes have attracted considerable attention because of their striking advantages in small-molecule separation. The preparation of an integrated MOF membrane is still a major challenge. Depositing a uniform seed layer on a support for secondary growth is a main route to obtaining an integrated MOF membrane. A novel seeding method to prepare HKUST-1 (known as Cu(3)(btc)(2)) membranes on porous α-alumina supports is reported. The in situ production of the seed layer was realized in step-by-step fashion via the coordination of H(3)btc and Cu(2+) on an α-alumina support. The formation process of the seed layer was observed by ultraviolet-visible absorption spectroscopy and atomic force microscopy. An integrated HKUST-1 membrane could be synthesized by the secondary hydrothermal growth on the seeded support. The gas permeation performance of the membrane was evaluated. © 2011 American Chemical Society

Integrated Microbial Electrolysis Cell (MEC) with an anaerobic Membrane Bioreactor (MBR) for low strength wastewater treatment, energy harvesting and water reclamation

KAUST Repository

Jimenez Sandoval, Rodrigo J.

2013-11-01

Shortage of potable water is a problem that affects many nations in the world and it will aggravate in a near future if pertinent actions are not carried out. Decrease in consumption, improvements in water distribution systems to avoid losses and more efficient water treatment processes are some actions that can be implemented to attack this problem. Membrane technology and biological processes are used in wastewater treatment to achieve high water quality standards. Some other technologies, besides water treatment, attempt to obtain energy from organic wastes present in water. In this study, a proof-of-concept was accomplished demonstrating that a Microbial Electrolysis Cell can be fully integrated with a Membrane Bioreactor to achieve wastewater treatment and harvest energy. Conductive hollow fiber membranes made of nickel functioned as both filter material for treated water reclamation and as a cathode to catalyze hydrogen production reaction. The produced hydrogen was subsequently converted into methane by hydrogenotrophic methanogens. Organic removal was 98.9% irrespective of operation mode. Maximum volumetric hydrogen production rate was 0.2 m3/m3d, while maximum current density achieved was 6.1 A/m2 (based on cathode surface area). Biofouling, an unavoidable phenomenon in traditional MBRs, can be minimized in this system through self-cleaning approach of hybrid membranes by hydrogen production. The increased rate of hydrogen evolution at high applied voltage (0.9 V) reduces the membrane fouling. Improvements can be done in the system to make it as a promising net energy positive technology for the low strength wastewater treatment.

Lactobacillus casei combats acid stress by maintaining cell membrane functionality.

Science.gov (United States)

Wu, Chongde; Zhang, Juan; Wang, Miao; Du, Guocheng; Chen, Jian

2012-07-01

Lactobacillus casei strains have traditionally been recognized as probiotics and frequently used as adjunct culture in fermented dairy products where lactic acid stress is a frequently encountered environmental condition. We have investigated the effect of lactic acid stress on the cell membrane of L. casei Zhang [wild type (WT)] and its acid-resistant mutant Lbz-2. Both strains were grown under glucose-limiting conditions in chemostats; following challenge by low pH, the cell membrane stress responses were investigated. In response to acid stress, cell membrane fluidity decreased and its fatty acid composition changed to reduce the damage caused by lactic acid. Compared with the WT, the acid-resistant mutant exhibited numerous survival advantages, such as higher membrane fluidity, higher proportions of unsaturated fatty acids, and higher mean chain length. In addition, cell integrity analysis showed that the mutant maintained a more intact cellular structure and lower membrane permeability after environmental acidification. These results indicate that alteration in membrane fluidity, fatty acid distribution, and cell integrity are common mechanisms utilized by L. casei to withstand severe acidification and to reduce the deleterious effect of lactic acid on the cell membrane. This detailed comparison of cell membrane responses between the WT and mutant add to our knowledge of the acid stress adaptation and thus enable new strategies to be developed aimed at improving the industrial performance of this species under acid stress.

Synthetic membrane-targeted antibiotics.

Science.gov (United States)

Vooturi, S K; Firestine, S M

2010-01-01

Antimicrobial resistance continues to evolve and presents serious challenges in the therapy of both nosocomial and community-acquired infections. The rise of resistant strains like methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA) and vancomycin-resistant enterococci (VRE) suggests that antimicrobial resistance is an inevitable evolutionary response to antimicrobial use. This highlights the tremendous need for antibiotics against new bacterial targets. Agents that target the integrity of bacterial membrane are relatively novel in the clinical armamentarium. Daptomycin, a lipopeptide is a classical example of membrane-bound antibiotic. Nature has also utilized this tactic. Antimicrobial peptides (AMPs), which are found in all kingdoms, function primarily by permeabilizing the bacterial membrane. AMPs have several advantages over existing antibiotics including a broad spectrum of activity, rapid bactericidal activity, no cross-resistance with the existing antibiotics and a low probability for developing resistance. Currently, a small number of peptides have been developed for clinical use but therapeutic applications are limited because of poor bioavailability and high manufacturing cost. However, their broad specificity, potent activity and lower probability for resistance have spurred the search for synthetic mimetics of antimicrobial peptides as membrane-active antibiotics. In this review, we will discuss the different classes of synthetic membrane-bound antibiotics published since 2004.

Transferable, conductive TiO{sub 2} nanotube membranes for optoelectronics

Energy Technology Data Exchange (ETDEWEB)

Liu, Guohua [School of Energy and Environment, Anhui University of Technology, Maanshan 243002 (China); Department of Micro and Nano Systems Technology, Vestfold University College, Horten 3184 (Norway); Chen, Ting [School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou 510275 (China); Sun, Yunlan; Chen, Guang [School of Energy and Environment, Anhui University of Technology, Maanshan 243002 (China); Wang, Kaiying, E-mail: Kaiying.Wang@hbv.no [Department of Micro and Nano Systems Technology, Vestfold University College, Horten 3184 (Norway)

2014-08-30

Graphical abstract: An optoelectronic device with vertical architecture offers straight conducting filaments for electron transportation. - Highlights: • Highly porous TiO{sub 2} nanotube membranes are prepared by two-step anodization. • An optoelectronic device is integrated with photocurrent transportation along the nanotube axial. • Straight conducting nano-filaments are beneficial for electron transportation. • Photoconductive performances are demonstrated under front/back-illumination. - Abstract: We report a facile approach for preparing free-standing and crystalline TiO{sub 2} nanotube membranes (TNMs) by taking advantage of differential mechanical stress between two anodic layers. The membrane exhibits visible light transmittance (∼40%) and UV absorption (∼99%) with good flexibility, which is favorable to integrate with substrates in optoelectronics. A sandwich-type device is assembled through stacking the membrane and substrates. The dependence of current-perpendicular-to-membrane vs applied voltage shows a remarkable photoconductive performance for both front and back illumination. The photocurrent value increases ∼2 or 3 orders magnitude under UV light radiation as compared to that in darkness. The photoresponse is arisen from high internal gain caused by hole trapping along the nanotube walls. This work is crucial for understanding intrinsic optical properties of nanostructured membranes.

Expression of fungal cutinase and swollenin in tobacco chloroplasts reveals novel enzyme functions and/or substrates.

Directory of Open Access Journals (Sweden)

Dheeraj Verma

Full Text Available In order to produce low-cost biomass hydrolyzing enzymes, transplastomic lines were generated that expressed cutinase or swollenin within chloroplasts. While swollenin expressing plants were homoplasmic, cutinase transplastomic lines remained heteroplasmic. Both transplastomic lines showed interesting modifications in their phenotype, chloroplast structure, and functions. Ultrastructural analysis of chloroplasts from cutinase- and swollenin-expressing plants did not show typical lens shape and granal stacks. But, their thylakoid membranes showed unique scroll like structures and chloroplast envelope displayed protrusions, stretching into the cytoplasm. Unusual honeycomb structures typically observed in etioplasts were observed in mature chloroplasts expressing swollenin. Treatment of cotton fiber with chloroplast-derived swollenin showed enlarged segments and the intertwined inner fibers were irreversibly unwound and fully opened up due to expansin activity of swollenin, causing disruption of hydrogen bonds in cellulose fibers. Cutinase transplastomic plants showed esterase and lipase activity, while swollenin transplastomic lines lacked such enzyme activities. Higher plants contain two major galactolipids, monogalactosyldiacylglycerol (MGDG and digalactosyldiacylglycerol (DGDG, in their chloroplast thylakoid membranes that play distinct roles in their structural organization. Surprisingly, purified cutinase effectively hydrolyzed DGDG to MGDG, showing alpha galactosidase activity. Such hydrolysis resulted in unstacking of granal thylakoids in chloroplasts and other structural changes. These results demonstrate DGDG as novel substrate and function for cutinase. Both MGDG and DGDG were reduced up to 47.7% and 39.7% in cutinase and 68.5% and 67.5% in swollenin expressing plants. Novel properties and functions of both enzymes reported here for the first time should lead to better understanding and enhanced biomass hydrolysis.

Photosystem II function and dynamics in three widely used Arabidopsis thaliana accessions.

Directory of Open Access Journals (Sweden)

Lan Yin

Full Text Available Columbia-0 (Col-0, Wassilewskija-4 (Ws-4, and Landsberg erecta-0 (Ler-0 are used as background lines for many public Arabidopsis mutant collections, and for investigation in laboratory conditions of plant processes, including photosynthesis and response to high-intensity light (HL. The photosystem II (PSII complex is sensitive to HL and requires repair to sustain its function. PSII repair is a multistep process controlled by numerous factors, including protein phosphorylation and thylakoid membrane stacking. Here we have characterized the function and dynamics of PSII complex under growth-light and HL conditions. Ws-4 displayed 30% more thylakoid lipids per chlorophyll and 40% less chlorophyll per carotenoid than Col-0 and Ler-0. There were no large differences in thylakoid stacking, photoprotection and relative levels of photosynthetic complexes among the three accessions. An increased efficiency of PSII closure was found in Ws-4 following illumination with saturation flashes or continuous light. Phosphorylation of the PSII D1/D2 proteins was reduced by 50% in Ws-4 as compared to Col-0 and Ler-0. An increase in abundance of the responsible STN8 kinase in response to HL treatment was found in all three accessions, but Ws-4 displayed 50% lower levels than Col-0 and Ler-0. Despite this, the HL treatment caused in Ws-4 the lagest extent of PSII inactivation, disassembly, D1 protein degradation, and the largest decrease in the size of stacked thylakoids. The dilution of chlorophyll-protein complexes with additional lipids and carotenoids in Ws-4 may represent a mechanism to facilitate lateral protein traffic in the membrane, thus compensating for the lack of a full complement of STN8 kinase. Nevertheless, additional PSII damage occurs in Ws-4, which exceeds the D1 protein synthesis capacity, thus leading to enhanced photoinhibition. Our findings are valuable for selection of appropriate background line for PSII characterization in Arabidopsis

Role of membranes and membrane reactors in the hydrogen supply of fuel cells for transports

Energy Technology Data Exchange (ETDEWEB)

Julbe, A.; Guizard, Ch. [Institut Europeen des Membranes, UMII, Lab. des Materiaux et des Procedes Membranaires, CNRS UMR 5635, 34 - Montpellier (France)

2000-07-01

Production, storage and supply of high-purity hydrogen as a clean and efficient fuel is central to fuel cells technology, in particular in vehicle traction. Actually, technologies for handling liquefied or gaseous hydrogen in transports are not available so that a number of alternative fuels are considered with the aim of in-situ generation of hydrogen through catalytic processes. The integrated concept of membrane reactors (MRs) can greatly benefit to these technologies. Particular emphasis is put on inorganic membranes and their role in MRs performance for H{sub 2} production.

Facile fabrication of Ag dendrite-integrated anodic aluminum oxide membrane as effective three-dimensional SERS substrate

Science.gov (United States)

Zhang, Cong-yun; Lu, Ya; Zhao, Bin; Hao, Yao-wu; Liu, Ya-qing

2016-07-01

A novel surface enhanced Raman scattering (SERS)-active substrate has been successfully developed, where Ag-dendrites are assembled on the surface and embedded in the channels of anodic aluminum oxide (AAO) membrane, via electrodeposition in AgNO3/PVP aqueous system. Reaction conditions were systematically investigated to attain the best Raman enhancement. The growth mechanism of Ag dendritic nanostructures has been proposed. The Ag dendrite-integrated AAO membrane with unique hierarchical structures exhibits high SERS activity for detecting rhodamine 6G with a detection limit as low as 1 × 10-11 M. Furthermore, the three-dimensional (3D) substrates display a good reproducibility with the average intensity variations at the major Raman peak less than 12%. Most importantly, the 3D SERS substrates without any surface modification show an outstanding SERS response for the molecules with weak affinity for noble metal surfaces. The potential application for the detection of polycyclic aromatic hydrocarbons (PAHs) was evaluated with fluoranthene as Raman target molecule and a sensitive SERS detection with a limit down to 10-8 M was reached. The 3D SERS-active substrate shows promising potential for rapid detection of trace organic pollutants even weak affinity molecules in the environment.

Integrative Analysis of Subcellular Quantitative Proteomics Studies Reveals Functional Cytoskeleton Membrane-Lipid Raft Interactions in Cancer.

Science.gov (United States)

Shah, Anup D; Inder, Kerry L; Shah, Alok K; Cristino, Alexandre S; McKie, Arthur B; Gabra, Hani; Davis, Melissa J; Hill, Michelle M

2016-10-07

Lipid rafts are dynamic membrane microdomains that orchestrate molecular interactions and are implicated in cancer development. To understand the functions of lipid rafts in cancer, we performed an integrated analysis of quantitative lipid raft proteomics data sets modeling progression in breast cancer, melanoma, and renal cell carcinoma. This analysis revealed that cancer development is associated with increased membrane raft-cytoskeleton interactions, with ∼40% of elevated lipid raft proteins being cytoskeletal components. Previous studies suggest a potential functional role for the raft-cytoskeleton in the action of the putative tumor suppressors PTRF/Cavin-1 and Merlin. To extend the observation, we examined lipid raft proteome modulation by an unrelated tumor suppressor opioid binding protein cell-adhesion molecule (OPCML) in ovarian cancer SKOV3 cells. In agreement with the other model systems, quantitative proteomics revealed that 39% of OPCML-depleted lipid raft proteins are cytoskeletal components, with microfilaments and intermediate filaments specifically down-regulated. Furthermore, protein-protein interaction network and simulation analysis showed significantly higher interactions among cancer raft proteins compared with general human raft proteins. Collectively, these results suggest increased cytoskeleton-mediated stabilization of lipid raft domains with greater molecular interactions as a common, functional, and reversible feature of cancer cells.

A novel integrated thermal-/membrane-based solar energy-driven hybrid desalination system: Concept description and simulation results.

Science.gov (United States)

Kim, Young-Deuk; Thu, Kyaw; Ng, Kim Choon; Amy, Gary L; Ghaffour, Noreddine

2016-09-01

In this paper, a hybrid desalination system consisting of vacuum membrane distillation (VMD) and adsorption desalination (AD) units, designated as VMD-AD cycle, is proposed. The synergetic integration of the VMD and AD is demonstrated where a useful effect of the AD cycle is channelled to boost the operation of the VMD process, namely the low vacuum environment to maintain the high pressure gradient across the microporous hydrophobic membrane. A solar-assisted multi-stage VMD-AD hybrid desalination system with temperature modulating unit is first designed, and its performance is then examined with a mathematical model of each component in the system and compared with the VMD-only system with temperature modulating and heat recovery units. The total water production and water recovery ratio of a solar-assisted 24-stage VMD-AD hybrid system are found to be about 21% and 23% higher, respectively, as compared to the VMD-only system. For the solar-assisted 24-stage VMD-AD desalination system having 150 m(2) of evacuated-tube collectors and 10 m(3) seawater storage tanks, both annual collector efficiency and solar fraction are close to 60%. Copyright © 2016 Elsevier Ltd. All rights reserved.

A novel integrated thermal-/membrane-based solar energy-driven hybrid desalination system: Concept description and simulation results

KAUST Repository

Kim, Youngdeuk

2016-05-03

In this paper, a hybrid desalination system consisting of vacuum membrane distillation (VMD) and adsorption desalination (AD) units, designated as VMD-AD cycle, is proposed. The synergetic integration of the VMD and AD is demonstrated where a useful effect of the AD cycle is channelled to boost the operation of the VMD process, namely the low vacuum environment to maintain the high pressure gradient across the microporous hydrophobic membrane. A solar-assisted multi-stage VMD-AD hybrid desalination system with temperature modulating unit is first designed, and its performance is then examined with a mathematical model of each component in the system and compared with the VMD-only system with temperature modulating and heat recovery units. The total water production and water recovery ratio of a solar-assisted 24-stage VMD-AD hybrid system are found to be about 21% and 23% higher, respectively, as compared to the VMD-only system. For the solar-assisted 24-stage VMD-AD desalination system having 150 m2 of evacuated-tube collectors and 10 m3 seawater storage tanks, both annual collector efficiency and solar fraction are close to 60%.

An investigation of desalination by nanofiltration, reverse osmosis and integrated (hybrid NF/RO) membranes employed in brackish water treatment.

Science.gov (United States)

Talaeipour, M; Nouri, J; Hassani, A H; Mahvi, A H

2017-01-01

hybrid system of those parameters and ions included salinity 78.65, TDS 76.52, EC 76.42, Cl 63.95, and Na 70.91. Comparing rejection percent in three above-mentioned methods, it could be concluded that, in reverse osmosis process, ions and non-ion parameters rejection ability were rather better than nanofiltration process, and also better in hybrid compared to reverse osmosis process. The results reported in this paper indicate that the integration of membrane nanofiltration with reverse osmosis (hybrid NF/RO) can be completed by each other probably to remove salinity, TDS, EC, Cl, and Na.

Iron Deprivation Affects Drug Susceptibilities of Mycobacteria Targeting Membrane Integrity

Directory of Open Access Journals (Sweden)

Rahul Pal

2015-01-01

Full Text Available Multidrug resistance (MDR acquired by Mycobacterium tuberculosis (MTB through continuous deployment of antitubercular drugs warrants immediate search for novel targets and mechanisms. The ability of MTB to sense and become accustomed to changes in the host is essential for survival and confers the basis of infection. A crucial condition that MTB must surmount is iron limitation, during the establishment of infection, since iron is required by both bacteria and humans. This study focuses on how iron deprivation affects drug susceptibilities of known anti-TB drugs in Mycobacterium smegmatis, a “surrogate of MTB.” We showed that iron deprivation leads to enhanced potency of most commonly used first line anti-TB drugs that could be reverted upon iron supplementation. We explored that membrane homeostasis is disrupted upon iron deprivation as revealed by enhanced membrane permeability and hypersensitivity to membrane perturbing agent leading to increased passive diffusion of drug and TEM images showing detectable differences in cell envelope thickness. Furthermore, iron seems to be indispensable to sustain genotoxic stress suggesting its possible role in DNA repair machinery. Taken together, we for the first time established a link between cellular iron and drug susceptibility of mycobacteria suggesting iron as novel determinant to combat MDR.

Ceramic membrane defouling (cleaning) by air Nano Bubbles.

Science.gov (United States)

Ghadimkhani, Aliasghar; Zhang, Wen; Marhaba, Taha

2016-03-01

Ceramic membranes are among the most promising technologies for membrane applications, owing to their excellent resistance to mechanical, chemical, and thermal stresses. However, membrane fouling is still an issue that hampers the applications at large scales. Air Nano Bubbles (NBs), due to high mass transfer efficiency, could potentially prevent fouling of ceramic membrane filtration processes. In this study, bench and pilot scale ceramic membrane filtration was performed with air NBs to resist fouling. To simulate fouling, humic acid, as an organic foulant, was applied to the membrane flat sheet surface. Complete membrane clogging was achieved in less than 6 h. Membrane defouling (cleaning) was performed by directly feeding of air NBs to the membrane cells. The surface of the ceramic membrane was superbly cleaned by air NBs, as revealed by atomic force microscope (AFM) images before and after the treatment. The permeate flux recovered to its initial level (e.g., 26.7 × 10(-9) m(3)/m(2)/s at applied pressure of 275.8 kPa), which indicated that NBs successfully unclogged the pores of the membrane. The integrated ceramic membrane and air NBs system holds potential as an innovative sustainable technology. Copyright © 2015 Elsevier Ltd. All rights reserved.

Integrating nanohybrid membranes of reduced graphene oxide: chitosan: silica sol gel with fiber optic SPR for caffeine detection

Science.gov (United States)

Kant, Ravi; Tabassum, Rana; Gupta, Banshi D.

2017-05-01

Caffeine is the most popular psychoactive drug consumed in the world for improving alertness and enhancing wakefulness. However, caffeine consumption beyond limits can result in lot of physiological complications in human beings. In this work, we report a novel detection scheme for caffeine integrating nanohybrid membranes of reduced graphene oxide (rGO) in chitosan modified silica sol gel (rGO: chitosan: silica sol gel) with fiber optic surface plasmon resonance. The chemically synthesized nanohybrid membrane forming the sensing route has been dip coated over silver coated unclad central portion of an optical fiber. The sensor works on the mechanism of modification of dielectric function of sensing layer on exposure to analyte solution which is manifested in terms of red shift in resonance wavelength. The concentration of rGO in polymer network of chitosan and silica sol gel and dipping time of the silver coated probe in the solution of nanohybrid membrane have been optimized to extricate the supreme performance of the sensor. The optimized sensing probe possesses a reasonably good sensitivity and follows an exponentially declining trend within the entire investigating range of caffeine concentration. The sensor boasts of an unparalleled limit of detection value of 1.994 nM and works well in concentration range of 0-500 nM with a response time of 16 s. The impeccable sensor methodology adopted in this work combining fiber optic SPR with nanotechnology furnishes a novel perspective for caffeine determination in commercial foodstuffs and biological fluids.

Large-Aperture Membrane Active Phased-Array Antennas

Science.gov (United States)

Karasik, Boris; McGrath, William; Leduc, Henry

2009-01-01

Large-aperture phased-array microwave antennas supported by membranes are being developed for use in spaceborne interferometric synthetic aperture radar systems. There may also be terrestrial uses for such antennas supported on stationary membranes, large balloons, and blimps. These antennas are expected to have areal mass densities of about 2 kg/sq m, satisfying a need for lightweight alternatives to conventional rigid phased-array antennas, which have typical areal mass densities between 8 and 15 kg/sq m. The differences in areal mass densities translate to substantial differences in total mass in contemplated applications involving aperture areas as large as 400 sq m. A membrane phased-array antenna includes patch antenna elements in a repeating pattern. All previously reported membrane antennas were passive antennas; this is the first active membrane antenna that includes transmitting/receiving (T/R) electronic circuits as integral parts. Other integral parts of the antenna include a network of radio-frequency (RF) feed lines (more specifically, a corporate feed network) and of bias and control lines, all in the form of flexible copper strip conductors on flexible polymeric membranes. Each unit cell of a prototype antenna (see Figure 1) contains a patch antenna element and a compact T/R module that is compatible with flexible membrane circuitry. There are two membrane layers separated by a 12.7-mm air gap. Each membrane layer is made from a commercially available flexible circuit material that, as supplied, comprises a 127-micron-thick polyimide dielectric layer clad on both sides with 17.5-micron-thick copper layers. The copper layers are patterned into RF, bias, and control conductors. The T/R module is located on the back side of the ground plane and is RF-coupled to the patch element via a slot. The T/R module is a hybrid multilayer module assembled and packaged independently and attached to the membrane array. At the time of reporting the information for

Characterization of membrane association of Rinderpest virus matrix protein

International Nuclear Information System (INIS)

Subhashri, R.; Shaila, M.S.

2007-01-01

Paramyxovirus matrix protein is believed to play a crucial role in the assembly and maturation of the virus particle by bringing the major viral components together at the budding site in the host cell. The membrane association capability of many enveloped virus matrix proteins has been characterized to be their intrinsic property. In this work, we have characterized the membrane association of Rinderpest virus matrix (M) protein. The M protein of Rinderpest virus when expressed in the absence of other viral proteins is present both in the cytoplasm and plasma membrane. When expressed as GFP fusion protein, the M protein gets localized into plasma membrane protrusions. High salt and alkaline conditions resulted in partial dissociation of M protein from cell membrane. Thus, M protein behaves like an integral membrane protein although its primary structure suggests it to be a peripheral membrane protein

Lipid corralling and poloxamer squeeze-out in membranes

DEFF Research Database (Denmark)

Wu, G.H.; Majewski, J.; Ege, C.

2004-01-01

Using x-ray scattering measurements we have quantitatively determined the effect of poloxamer 188 (P188), a polymer known to seal damaged membranes, on the structure of lipid monolayers. P188 selectively inserts into low lipid-density regions of the membrane and "corrals" lipid molecules to pack...... tightly, leading to unexpected Bragg peaks at low nominal lipid density and inducing lipid/poloxamer phase separation. At tighter lipid packing, the once inserted P188 is squeezed out, allowing the poloxamer to gracefully exit when the membrane integrity is restored....

Linking Cellular Mechanisms to Behavior: Entorhinal Persistent Spiking and Membrane Potential Oscillations May Underlie Path Integration, Grid Cell Firing, and Episodic Memory

Directory of Open Access Journals (Sweden)

Michael E. Hasselmo

2008-01-01

Full Text Available The entorhinal cortex plays an important role in spatial memory and episodic memory functions. These functions may result from cellular mechanisms for integration of the afferent input to entorhinal cortex. This article reviews physiological data on persistent spiking and membrane potential oscillations in entorhinal cortex then presents models showing how both these cellular mechanisms could contribute to properties observed during unit recording, including grid cell firing, and how they could underlie behavioural functions including path integration. The interaction of oscillations and persistent firing could contribute to encoding and retrieval of trajectories through space and time as a mechanism relevant to episodic memory.

Polymeric membranes for guided bone regeneration.

Science.gov (United States)

Gentile, Piergiorgio; Chiono, Valeria; Tonda-Turo, Chiara; Ferreira, Ana M; Ciardelli, Gianluca

2011-10-01

In this review, different barrier membranes for guided bone regeneration (GBR) are described as a useful surgical technique to enhance bone regeneration in damaged alveolar sites before performing implants and fitting other dental appliances. The GBR procedure encourages bone regeneration through cellular exclusion and avoids the invasion of epithelial and connective tissues that grow at the defective site instead of bone tissue. The barrier membrane should satisfy various properties, such as biocompatibility, non-immunogenicity, non-toxicity, and a degradation rate that is long enough to permit mechanical support during bone formation. Other characteristics such as tissue integration, nutrient transfer, space maintenance and manageability are also of interest. In this review, various non-resorbable and resorbable commercially available membranes are described, based on expanded polytetrafluoroethylene, poly(lactic acid), poly(glycolic acid) and their copolymers. The polyester-based membranes are biodegradable, permit a single-stage procedure, and have higher manageability than non-resorbable membranes; however, they have shown poor biocompatibility. In contrast, membranes based on natural materials, such as collagen, are biocompatible but are characterized by poor mechanical properties and stability due to their early degradation. Moreover, new approaches are described, such as the use of multi-layered, graft-copolymer-based and composite membranes containing osteoconductive ceramic fillers as alternatives to conventional membranes. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Stability of integral membrane proteins under high hydrostatic pressure: the LH2 and LH3 antenna pigment-protein complexes from photosynthetic bacteria.

Science.gov (United States)

Kangur, Liina; Timpmann, Kõu; Freiberg, Arvi

2008-07-03

The bacteriochlorophyll a-containing LH2 and LH3 antenna complexes are the integral membrane proteins that catalyze the photosynthetic process in purple photosynthetic bacteria. The LH2 complex from Rhodobacter sphaeroides shows characteristic strong absorbance at 800 and 850 nm due to the pigment molecules confined in two separate areas of the protein. In the LH3 complex from Rhodopesudomonas acidophila the corresponding bands peak at 800 and 820 nm. Using the bacteriochlorophyll a cofactors as intrinsic probes to monitor local changes in the protein structure, we investigate spectral responses of the antenna complexes to very high hydrostatic pressures up to 2.5 GPa when embedded into natural membrane environment or extracted with detergent. We first demonstrate that high pressure does induce significant alterations to the tertiary structure of the proteins not only in proximity of the 800 nm-absorbing bacteriochlorophyll a molecules known previously (Gall, A.; et al. Biochemistry 2003, 42, 13019) but also of the 850 nm- and 820 nm-absorbing molecules, including breakage of the hydrogen bond they are involved in. The membrane-protected complexes appear more resilient to damaging effects of the compression compared with the complexes extracted into mixed detergent-buffer environment. Increased resistance of the isolated complexes is observed at high protein concentration resulting aggregation as well as when cosolvent (glycerol) is added into the solution. These stability variations correlate with ability of penetration of the surrounding polar solvent (water) into the hydrophobic protein interiors, being thus the principal reason of the pressure-induced denaturation of the proteins. Considerable variability of elastic properties of the isolated complexes was also observed, tentatively assigned to heterogeneous protein packing in detergent micelles. While a number of the isolated complexes release most of their bacteriochlorophyll a content under high pressure

Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase.

Science.gov (United States)

Hatahet, Feras; Blazyk, Jessica L; Martineau, Eugenie; Mandela, Eric; Zhao, Yongxin; Campbell, Robert E; Beckwith, Jonathan; Boyd, Dana

2015-12-08

Functional overexpression of polytopic membrane proteins, particularly when in a foreign host, is often a challenging task. Factors that negatively affect such processes are poorly understood. Using the mammalian membrane protein vitamin K epoxide reductase (VKORc1) as a reporter, we describe a genetic selection approach allowing the isolation of Escherichia coli mutants capable of functionally expressing this blood-coagulation enzyme. The isolated mutants map to components of membrane protein assembly and quality control proteins YidC and HslV. We show that changes in the VKORc1 sequence and in the YidC hydrophilic groove along with the inactivation of HslV promote VKORc1 activity and dramatically increase its expression level. We hypothesize that such changes correct for mismatches in the membrane topogenic signals between E. coli and eukaryotic cells guiding proper membrane integration. Furthermore, the obtained mutants allow the study of VKORc1 reaction mechanisms, inhibition by warfarin, and the high-throughput screening for potential anticoagulants.

A difference gel electrophoresis study on thylakoids isolated from poplar leaves reveals a negative impact of ozone exposure on membrane proteins.

Science.gov (United States)

Bohler, Sacha; Sergeant, Kjell; Hoffmann, Lucien; Dizengremel, Pierre; Hausman, Jean-Francois; Renaut, Jenny; Jolivet, Yves

2011-07-01

Populus tremula L. x P. alba L. (Populus x canescens (Aiton) Smith), clone INRA 717-1-B4, saplings were subjected to 120 ppb ozone exposure for 28 days. Chloroplasts were isolated, and the membrane proteins, solubilized using the detergent 1,2-diheptanoyl-sn-glycero-3-phosphocholine (DHPC), were analyzed in a difference gel electrophoresis (DiGE) experiment comparing control versus ozone-exposed plants. Extrinsic photosystem (PS) proteins and adenosine triphosphatase (ATPase) subunits were detected to vary in abundance. The general trend was a decrease in abundance, except for ferredoxin-NADP(+) oxidoreductase (FNR), which increased after the first 7 days of exposure. The up-regulation of FNR would increase NAPDH production for reducing power and detoxification inside and outside of the chloroplast. Later on, FNR and a number of PS and ATPase subunits decrease in abundance. This could be the result of oxidative processes on chloroplast proteins but could also be a way to down-regulate photochemical reactions in response to an inhibition in Calvin cycle activity.

Isolation of monodisperse nanodisc-reconstituted membrane proteins using free flow electrophoresis

DEFF Research Database (Denmark)

Justesen, Bo Højen; Laursen, Tomas; Weber, Gerhard

2013-01-01

Free flow electrophoresis is used for rapid and high-recovery isolation of homogeneous preparations of functionally active membrane proteins inserted into nanodiscs. The approach enables isolation of integral and membrane anchored proteins and is also applicable following introduction of, e...

Self-assembled tethered bimolecular lipid membranes.

Science.gov (United States)

Sinner, Eva-Kathrin; Ritz, Sandra; Naumann, Renate; Schiller, Stefan; Knoll, Wolfgang

2009-01-01

This chapter describes some of the strategies developed in our group for designing, constructing and structurally and functionally characterizing tethered bimolecular lipid membranes (tBLM). We introduce this platform as a novel model membrane system that complements the existing ones, for example, Langmuir monolayers, vesicular liposomal dispersions and bimolecular ("black") lipid membranes. Moreover, it offers the additional advantage of allowing for studies of the influence of membrane structure and order on the function of integral proteins, for example, on how the composition and organization of lipids in a mixed membrane influence the ion translocation activity of integral channel proteins. The first strategy that we introduce concerns the preparation of tethered monolayers by the self-assembly of telechelics. Their molecular architecture with a headgroup, a spacer unit (the "tether") and the amphiphile that mimics the lipid molecule allows them to bind specifically to the solid support thus forming the proximal layer of the final architecture. After fusion of vesicles that could contain reconstituted proteins from a liposomal dispersion in contact to this monolayer the tethered bimolecular lipid membrane is obtained. This can then be characterized by a broad range of surface analytical techniques, including surface plasmon spectroscopies, the quartz crystal microbalance, fluorescence and IR spectroscopies, and electrochemical techniques, to mention a few. It is shown that this concept allows for the construction of tethered lipid bilayers with outstanding electrical properties including resistivities in excess of 10 MOmega cm2. A modified strategy uses the assembly of peptides as spacers that couple covalently via their engineered sulfhydryl or lipoic acid groups at the N-terminus to the employed gold substrate, while their C-terminus is being activated afterward for the coupling of, for example, dimyristoylphosphatidylethanol amine (DMPE) lipid molecules

Final Report - Membranes and MEA's for Dry, Hot Operating Conditions

Energy Technology Data Exchange (ETDEWEB)

Hamrock, Steven J

2011-06-30

The focus of this program was to develop a new Proton Exchange Membrane (PEM) which can operate under hotter, dryer conditions than the state of the art membranes today and integrate it into a Membrane Electrode Assembly (MEA). These MEA's should meet the performance and durability requirements outlined in the solicitation, operating under low humidification conditions and at temperatures ranging from -20ºC to 120ºC, to meet 2010 DOE technical targets for membranes. This membrane should operate under low humidification conditions and at temperatures ranging from -20ºC to 120ºC in order to meet DOE HFCIT 2010 commercialization targets for automotive fuel cells. Membranes developed in this program may also have improved durability and performance characteristics making them useful in stationary fuel cell applications. The new membranes, and the MEA's comprising them, should be manufacturable at high volumes and at costs which can meet industry and DOE targets. This work included: A) Studies to better understand factors controlling proton transport within the electrolyte membrane, mechanisms of polymer degradation (in situ and ex situ) and membrane durability in an MEA; B) Development of new polymers with increased proton conductivity over the range of temperatures from -20ºC to 120ºC and at lower levels of humidification and with improved chemical and mechanical stability; C) Development of new membrane additives for increased durability and conductivity under these dry conditions; D) Integration of these new materials into membranes and membranes into MEA's, including catalyst and gas diffusion layer selection and integration; E) Verification that these materials can be made using processes which are scalable to commercial volumes using cost effective methods; F) MEA testing in single cells using realistic automotive testing protocols. This project addresses technical barriers A (Durability) and C (Performance) from the Fuel Cells section of the

Simultaneous nitrogen and organics removal using membrane aeration and effluent ultrafiltration in an anaerobic fluidized membrane bioreactor

KAUST Repository

Ye, Yaoli; Saikaly, Pascal; Logan, B.E.

2017-01-01

Dissolved methane and a lack of nutrient removal are two concerns for treatment of wastewater using anaerobic fluidized bed membrane bioreactors (AFMBRs). Membrane aerators were integrated into an AFMBR to form an Aeration membrane fluidized bed membrane bioreactor (AeMFMBR) capable of simultaneous removal of organic matter and ammonia without production of dissolved methane. Good effluent quality was obtained with no detectable suspended solids, 93±5% of chemical oxygen demand (COD) removal to 14±11 mg/L, and 74±8% of total ammonia (TA) removal to 12±3 mg-N/L for domestic wastewater (COD of 193±23 mg/L and TA of 49±5 mg-N/L) treatment. Nitrate and nitrite concentrations were always low (< 1 mg-N/L) during continuous flow treatment. Membrane fouling was well controlled by fluidization of the granular activated carbon (GAC) particles (transmembrane pressures maintained <3 kPa). Analysis of the microbial communities suggested that nitrogen removal was due to nitrification and denitrification based on the presence of microorganisms associated with these processes.

Simultaneous nitrogen and organics removal using membrane aeration and effluent ultrafiltration in an anaerobic fluidized membrane bioreactor

KAUST Repository

Ye, Yaoli

2017-08-03

Dissolved methane and a lack of nutrient removal are two concerns for treatment of wastewater using anaerobic fluidized bed membrane bioreactors (AFMBRs). Membrane aerators were integrated into an AFMBR to form an Aeration membrane fluidized bed membrane bioreactor (AeMFMBR) capable of simultaneous removal of organic matter and ammonia without production of dissolved methane. Good effluent quality was obtained with no detectable suspended solids, 93±5% of chemical oxygen demand (COD) removal to 14±11 mg/L, and 74±8% of total ammonia (TA) removal to 12±3 mg-N/L for domestic wastewater (COD of 193±23 mg/L and TA of 49±5 mg-N/L) treatment. Nitrate and nitrite concentrations were always low (< 1 mg-N/L) during continuous flow treatment. Membrane fouling was well controlled by fluidization of the granular activated carbon (GAC) particles (transmembrane pressures maintained <3 kPa). Analysis of the microbial communities suggested that nitrogen removal was due to nitrification and denitrification based on the presence of microorganisms associated with these processes.

Integration of computational modeling with membrane transport studies reveals new insights into amino acid exchange transport mechanisms

Science.gov (United States)

Widdows, Kate L.; Panitchob, Nuttanont; Crocker, Ian P.; Please, Colin P.; Hanson, Mark A.; Sibley, Colin P.; Johnstone, Edward D.; Sengers, Bram G.; Lewis, Rohan M.; Glazier, Jocelyn D.

2015-01-01

Uptake of system L amino acid substrates into isolated placental plasma membrane vesicles in the absence of opposing side amino acid (zero-trans uptake) is incompatible with the concept of obligatory exchange, where influx of amino acid is coupled to efflux. We therefore hypothesized that system L amino acid exchange transporters are not fully obligatory and/or that amino acids are initially present inside the vesicles. To address this, we combined computational modeling with vesicle transport assays and transporter localization studies to investigate the mechanisms mediating [14C]l-serine (a system L substrate) transport into human placental microvillous plasma membrane (MVM) vesicles. The carrier model provided a quantitative framework to test the 2 hypotheses that l-serine transport occurs by either obligate exchange or nonobligate exchange coupled with facilitated transport (mixed transport model). The computational model could only account for experimental [14C]l-serine uptake data when the transporter was not exclusively in exchange mode, best described by the mixed transport model. MVM vesicle isolates contained endogenous amino acids allowing for potential contribution to zero-trans uptake. Both L-type amino acid transporter (LAT)1 and LAT2 subtypes of system L were distributed to MVM, with l-serine transport attributed to LAT2. These findings suggest that exchange transporters do not function exclusively as obligate exchangers.—Widdows, K. L., Panitchob, N., Crocker, I. P., Please, C. P., Hanson, M. A., Sibley, C. P., Johnstone, E. D., Sengers, B. G., Lewis, R. M., Glazier, J. D. Integration of computational modeling with membrane transport studies reveals new insights into amino acid exchange transport mechanisms. PMID:25761365

Perturbations of carotenoid and tetrapyrrole biosynthetic pathways result in differential alterations in chloroplast function and plastid signaling.

Science.gov (United States)

Park, Joon-Heum; Jung, Sunyo

2017-01-22

In this study, we used the biosynthetic inhibitors of carotenoid and tetrapyrrole biosynthetic pathways, norflurazon (NF) and oxyfluorfen (OF), as tools to gain insight into mechanisms of photooxidation in rice plants. NF resulted in bleaching symptom on leaves of the treated plants, whereas OF treatment developed a fast symptom of an apparent necrotic phenotype. Both plants exhibited decreases in photosynthetic efficiency, as indicated by F v /F m . NF caused severe disruption in thylakoid membranes, whereas OF-treated plants exhibited disruption of chloroplast envelope and plasma membrane. Levels of Lhca and Lhcb proteins in photosystem I (PSI) and PSII were reduced by photooxidative stress in NF- and OF-treated plants, with a greater decrease in NF plants. The down-regulation of nuclear-encoded photosynthesis genes Lhcb and rbcS was also found in both NF- and OF-treated plants, whereas plastid-encoded photosynthetic genes including RbcL, PsaC, and PsbD accumulated normally in NF plants but decreased drastically in OF plants. This proposes that the plastids in NF plants retain their potential to develop thylakoid membranes and that photobleaching is mainly controlled by nuclear genes. Distinct photooxidation patterns between NF- and OF-treated plants developed differential signaling, which might enable the plant to coordinate the expression of photosynthetic genes from the nuclear and plastidic genomes. Copyright © 2016 Elsevier Inc. All rights reserved.

Biomimetic membranes for sensor and separation applications

CERN Document Server

2012-01-01

This book addresses the possibilities and challenges in mimicking biological membranes and creating membrane-based sensor and separation devices. It covers recent advances in developing biomimetic membranes for technological applications with a focus on the use of integral membrane protein mediated transport. It describes the fundamentals of biosensing as well as separation and shows how the two processes work together in biological systems. The book provides an overview of the current state of the art, points to areas that need further investigation and anticipates future directions in the field. Biomimetics is a truly cross-disciplinary approach and this is exemplified by the challenges in mimicking osmotic processes as they occur in nature using aquaporin protein water channels as central building blocks. In the development of a biomimetic sensor/separation technology, both channel and carrier proteins are important and examples of how these may be reconstituted and controlled in biomimetic membranes are ...

Functional dynamics of cell surface membrane proteins.

Science.gov (United States)

Nishida, Noritaka; Osawa, Masanori; Takeuchi, Koh; Imai, Shunsuke; Stampoulis, Pavlos; Kofuku, Yutaka; Ueda, Takumi; Shimada, Ichio

2014-04-01

Cell surface receptors are integral membrane proteins that receive external stimuli, and transmit signals across plasma membranes. In the conventional view of receptor activation, ligand binding to the extracellular side of the receptor induces conformational changes, which convert the structure of the receptor into an active conformation. However, recent NMR studies of cell surface membrane proteins have revealed that their structures are more dynamic than previously envisioned, and they fluctuate between multiple conformations in an equilibrium on various timescales. In addition, NMR analyses, along with biochemical and cell biological experiments indicated that such dynamical properties are critical for the proper functions of the receptors. In this review, we will describe several NMR studies that revealed direct linkage between the structural dynamics and the functions of the cell surface membrane proteins, such as G-protein coupled receptors (GPCRs), ion channels, membrane transporters, and cell adhesion molecules. Copyright © 2013 Elsevier Inc. All rights reserved.

An anaerobic membrane bioreactor - membrane distillation hybrid system for energy recovery and water reuse: Removal performance of organic carbon, nutrients, and trace organic contaminants.

Science.gov (United States)

Song, Xiaoye; Luo, Wenhai; McDonald, James; Khan, Stuart J; Hai, Faisal I; Price, William E; Nghiem, Long D

2018-07-01

In this study, a direct contact membrane distillation (MD) unit was integrated with an anaerobic membrane bioreactor (AnMBR) to simultaneously recover energy and produce high quality water for reuse from wastewater. Results show that AnMBR could produce 0.3-0.5L/g COD added biogas with a stable methane content of approximately 65%. By integrating MD with AnMBR, bulk organic matter and phosphate were almost completely removed. The removal of the 26 selected trace organic contaminants by AnMBR was compound specific, but the MD process could complement AnMBR removal, leading to an overall efficiency from 76% to complete removal by the integrated system. The results also show that, due to complete retention, organic matter (such as humic-like and protein-like substances) and inorganic salts accumulated in the MD feed solution and therefore resulted in significant fouling of the MD unit. As a result, the water flux of the MD process decreased continuously. Nevertheless, membrane pore wetting was not observed throughout the operation. Crown Copyright © 2018. Published by Elsevier B.V. All rights reserved.

Integrated test plan for the field demonstration of the supported liquid membrane unit

International Nuclear Information System (INIS)

Dunks, K.L.; Hodgson, K.M.

1995-06-01

This Integrated Test Plan describes the operation and testing of a hybrid reverse osmosis (RO)/coupled transport (CT) groundwater remediation test unit, also referred to as the Environmental Restoration Technology Demonstrations at the Hanford Site. The SLM will be used to remove uranium, technetium-99, and nitrate from a selected groundwater source at the Hanford Site. The overall purpose of this test is to determine the efficiency of the RO/CT membranes operating in a hybrid unit, the ease of operating and maintaining the SLM, and the amount of secondary waste generated as a result of processing. The goal of the SLM is to develop a RO/CT process that will be applicable for removing contaminants from almost any contaminated water. This includes the effluents generated as part of the day-to-day operation of most any US Department of Energy (DOE) site. The removal of contaminants from the groundwaters before they reach the Columbia River or offsite extraction wells will reduce the risk that the population will be exposed to these compounds and will reduce the cost of subsequent groundwater cleanup

Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

OpenAIRE

F. Cremonesi; A. Meucci; A. Lange-Consiglio

2013-01-01

The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19?C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using prop...

Hybrid membrane-microfluidic components using a novel ceramic MEMS technology

Science.gov (United States)

Lutz, Brent J.; Polyakov, Oleg; Rinaldo, Chris

2012-03-01

A novel hybrid nano/microfabrication technology has been employed to produce unique MEMS and microfluidic components that integrate nanoporous membranes. The components are made by micromachining a self-organized nanostructured ceramic material that is biocompatible and amenable to surface chemistry modification. Microfluidic structures, such as channels and wells, can be made with a precision of membranes can be integrated into the bottom of these structures, featuring a wide range of possible thicknesses, from 100 micron to membranes may be non-porous or porous (with controllable pore sizes from 200 nm to technology is highly scaleable, and thus can yield low-cost, reliable, disposable microcomponents and devices. Specific applications that can benefit from this technology includes cell culturing and assays, imaging by cryo-electron tomography, environmental sample processing, as well as many others.

On the mechanism of sulfite activation of chloroplast thylakoid ATPase and the relation of ADP tightly bound at a catalytic site to the binding change mechanism

Energy Technology Data Exchange (ETDEWEB)

Du, Z.; Boyer, P.D. (Univ. of California, Los Angeles (USA))

1990-01-16

Washed chloroplast thylakoid membranes upon exposure to ({sup 3}H)ADP retain in tightly bound ({sup 3}H)ADP on a catalytic site of the ATP synthase. The presence of sufficient endogenous or added Mg{sup 2+} results in an enzyme with essentially no ATPase activity. Sulfite activates the ATPase, and many molecules of ATP per synthase can be hydrolyzed before most of the bound ({sup 3}H)ADP is released, a result interpreted as indicating that the ADP is not bound at a site participating in catalysis by the sulfite-activated enzyme. The authors present evidence that this is not the case. The Mg{sup 2+}- and ADP-inhibited enzyme when exposed to MgATP and 20-100 mM sulfite shows a lag of about 1 min at 22{degree}C and of about 15 s at 37{degree}C before reaching the same steady-state rate as attained with light-activated ATPase that has not been inhibited by Mg{sup 2+} and ADP. The lag is not eliminated if the enzyme is exposed to sulfite prior to MgATP addition, indicating that ATPase turnover is necessary for the activation. The release of most of the bound ({sup 3}H)ADP parallels the onset of ATPase activity, although some ({sup 3}H)ADP is not released even with prolonged catalytic turnover and may be on poorly active or inactive enzyme or at noncatalytic sites. The results are consistent with most of the tightly bound ({sup 3}H)ADP being at a catalytic site and being replaced as this Mg{sup 2+}- and ADP-inhibited site regains equivalent participation with other catalytic sites on the activated enzyme. The sulfite activation can be explained by sulfite combination at a P{sub i} binding site of the enzyme-ADP-Mg{sup 2+} complex to give a form more readily activated by ATP binding at an alternative site.

Field-effect detection using phospholipid membranes -Topical Review

Directory of Open Access Journals (Sweden)

Chiho Kataoka-Hamai and Yuji Miyahara

2010-01-01

Full Text Available The application of field-effect devices to biosensors has become an area of intense research interest. An attractive feature of field-effect sensing is that the binding or reaction of biomolecules can be directly detected from a change in electrical signals. The integration of such field-effect devices into cell membrane mimics may lead to the development of biosensors useful in clinical and biotechnological applications. This review summarizes recent studies on the fabrication and characterization of field-effect devices incorporating model membranes. The incorporation of black lipid membranes and supported lipid monolayers and bilayers into semiconductor devices is described.

Integrative Analysis of the microRNAome and Transcriptome Illuminates the Response of Susceptible Rice Plants to Rice Stripe Virus.

Directory of Open Access Journals (Sweden)

Jian Yang

Full Text Available Rice stripe virus (RSV is one of the most serious rice viruses in East Asia. To investigate how rice responds to RSV infection, we integrated miRNA expression with parallel mRNA transcription profiling by deep sequencing. A total of 570 miRNAs were identified of which 69 miRNAs (56 up-regulated and 13 down-regulated were significantly modified by RSV infection. Digital gene expression (DGE analysis showed that 1274 mRNAs (431 up-regulated and 843 down-regulated genes were differentially expressed as a result of RSV infection. The differential expression of selected miRNAs and mRNAs was confirmed by qRT-PCR. Gene ontology (GO and pathway enrichment analysis showed that a complex set of miRNA and mRNA networks were selectively regulated by RSV infection. In particular, 63 differentially expressed miRNAs were found to be significantly and negatively correlated with 160 target mRNAs. Interestingly, 22 up-regulated miRNAs were negatively correlated with 24 down-regulated mRNAs encoding disease resistance-related proteins, indicating that the host defense responses were selectively suppressed by RSV infection. The suppression of both osa-miR1423-5p- and osa-miR1870-5p-mediated resistance pathways was further confirmed by qRT-PCR. Chloroplast functions were also targeted by RSV, especially the zeaxanthin cycle, which would affect the stability of thylakoid membranes and the biosynthesis of ABA. All these modifications may contribute to viral symptom development and provide new insights into the pathogenicity mechanisms of RSV.

EM Task 9 - Centrifugal membrane filtration

International Nuclear Information System (INIS)

Stepan, Daniel J.; Stevens, Bradley G.; Hetland, Melanie D.

1999-01-01

The overall project consists of several integrated research phases related to the applicability, continued development, demonstration, and commercialization of the SpinTek centrifugal membrane filtration process. Work performed during this reporting period consisted of Phase 2 evaluation of the SpinTek centrifugal membrane filtration technology and Phase 3, Technology Partnering. During Phase 1 testing conducted at the EERC using the SpinTek ST-IIL unit operating on a surrogate tank waste, a solids cake developed on the membrane surface. The solids cake was observed where linear membrane velocities were less than 17.5 ft/s and reduced the unobstructed membrane surface area up to 25%, reducing overall filtration performance. The primary goal of the Phase 2 research effort was to enhance filtration performance through the development and testing of alternative turbulence promoter designs. The turbulence promoters were designed to generate a shear force across the entire membrane surface sufficient to maintain a self-cleaning membrane capability and improve filtration efficiency and long-term performance. Specific Phase 2 research activities included the following: System modifications to accommodate an 11-in.-diameter, two-disk rotating membrane assembly; Development and fabrication of alternative turbulence promoter designs; Testing and evaluation of the existing and alternative turbulence promoters under selected operating conditions using a statistically designed test matrix; and Data reduction and analysis; The objective of Phase 3 research was to demonstrate the effectiveness of SpinTek's centrifugal membrane filtration as a pretreatment to remove suspended solids from a liquid waste upstream of 3M's WWL cartridge technology for the selective removal of technetium (Tc)

Models of dynamic extraction of lipid tethers from cell membranes

International Nuclear Information System (INIS)

Nowak, Sarah A; Chou, Tom

2010-01-01

When a ligand that is bound to an integral membrane receptor is pulled, the membrane and the underlying cytoskeleton can deform before either the membrane delaminates from the cytoskeleton or the ligand detaches from the receptor. If the membrane delaminates from the cytoskeleton, it may be further extruded and form a membrane tether. We develop a phenomenological model for this process by assuming that deformations obey Hooke's law up to a critical force at which the cell membrane locally detaches from the cytoskeleton and a membrane tether forms. We compute the probability of tether formation and show that tethers can be extruded only within an intermediate range of force loading rates and pulling velocities. The mean tether length that arises at the moment of ligand detachment is computed as are the force loading rates and pulling velocities that yield the longest tethers

CO₂ Capture Membrane Process for Power Plant Flue Gas

Energy Technology Data Exchange (ETDEWEB)

Toy, Lora [Research Triangle Inst. International, Research Triangle Park, NC (United States); Kataria, Atish [Research Triangle Inst. International, Research Triangle Park, NC (United States); Gupta, Raghubir [Research Triangle Inst. International, Research Triangle Park, NC (United States)

2012-04-01

�� permeance greater than 300 gas permeation units (GPU) targeted; - Development of next-generation polycarbonate hollow-fiber membranes and membrane modules with higher CO₂ permeance than current commercial polycarbonate membranes; - Development and fabrication of membrane hollow fibers and modules from candidate polymers; - Development of a CO₂ capture membrane process design and integration strategy suitable for end-of-pipe, retrofit installation; and - Techno-economic evaluation of the "best" integrated CO₂ capture membrane process design package In this report, the results of the project research and development efforts are discussed and include the post-combustion capture properties of the two membrane material platforms and the hollow-fiber membrane modules developed from them and the multi-stage process design and analysis developed for 90% CO₂ capture with 95% captured CO₂ purity.

Plants and fungi in the era of heterogeneous plasma membranes.

Science.gov (United States)

Opekarová, M; Malinsky, J; Tanner, W

2010-09-01

Examples from yeast and plant cells are described that show that their plasma membrane is laterally compartmented. Distinct lateral domains encompassing both specific lipids and integral proteins coexist within the plane of the plasma membrane. The compartments are either spatially stable and include distinct sets of proteins, or they are transiently formed to accomplish diverse functions. They are not related to lipid rafts or their clusters, as defined for mammalian cells. This review summarises only well-documented compartments of plasma membranes from plants and fungi, which have been recognised using microscopic approaches. In several cases, physiological functions of the membrane compartmentation are revealed.

Cholesterol asymmetry in synaptic plasma membranes.

Science.gov (United States)

Wood, W Gibson; Igbavboa, Urule; Müller, Walter E; Eckert, Gunter P

2011-03-01

Lipids are essential for the structural and functional integrity of membranes. Membrane lipids are not randomly distributed but are localized in different domains. A common characteristic of these membrane domains is their association with cholesterol. Lipid rafts and caveolae are examples of cholesterol enriched domains, which have attracted keen interest. However, two other important cholesterol domains are the exofacial and cytofacial leaflets of the plasma membrane. The two leaflets that make up the bilayer differ in their fluidity, electrical charge, lipid distribution, and active sites of certain proteins. The synaptic plasma membrane (SPM) cytofacial leaflet contains over 85% of the total SPM cholesterol as compared with the exofacial leaflet. This asymmetric distribution of cholesterol is not fixed or immobile but can be modified by different conditions in vivo: (i) chronic ethanol consumption; (ii) statins; (iii) aging; and (iv) apoE isoform. Several potential candidates have been proposed as mechanisms involved in regulation of SPM cholesterol asymmetry: apoE, low-density lipoprotein receptor, sterol carrier protein-2, fatty acid binding proteins, polyunsaturated fatty acids, P-glycoprotein and caveolin-1. This review examines cholesterol asymmetry in SPM, potential mechanisms of regulation and impact on membrane structure and function. © 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.

Hybrid Membrane/Absorption Process for Post-combustion CO2 Capture

Energy Technology Data Exchange (ETDEWEB)

Li, Shiguang; Shou, S.; Pyrzynski, Travis; Makkuni, Ajay; Meyer, Howard

2013-12-31

This report summarizes scientific/technical progress made for bench-scale membrane contactor technology for post-combustion CO2 capture from DOE Contract No. DE-FE-0004787. Budget Period 1 (BP1) membrane absorber, Budget Period 2 (BP2) membrane desorber and Budget Period 3 (BP3) integrated system and field testing studies have been completed successfully and met or exceeded the technical targets (≥ 90% CO2 removal and CO2 purity of 97% in one membrane stage). Significant breakthroughs are summarized below: BP1 research: The feasibility of utilizing the poly (ether ether ketone), PEEK, based hollow fiber contractor (HFC) in combination with chemical solvents to separate and capture at least 90% of the CO2 from simulated flue gases has been successfully established. Excellent progress has been made as we have achieved the BP1 goal: ≥ 1,000 membrane intrinsic CO2 permeance, ≥ 90% CO2 removal in one stage, ≤ 2 psi gas side pressure drop, and ≥ 1 (sec)-1 mass transfer coefficient. Initial test results also show that the CO2 capture performance, using activated Methyl Diethanol Amine (aMDEA) solvent, was not affected by flue gas contaminants O2 (~3%), NO2 (66 ppmv), and SO2 (145 ppmv). BP2 research: The feasibility of utilizing the PEEK HFC for CO2-loaded solvent regeneration has been successfully established High CO2 stripping flux, one order of magnitude higher than CO2 absorption flux, have been achieved. Refined economic evaluation based on BP1 membrane absorber and BP2 membrane desorber laboratory test data indicate that the CO2 capture costs are 36% lower than DOE’s benchmark amine absorption technology. BP3 research: A bench-scale system utilizing a membrane absorber and desorber was integrated into a continuous CO2 capture process using contactors containing 10 to 20 ft2 of membrane area. The integrated process operation was stable through a 100-hour laboratory test, utilizing a simulated flue gas stream. Greater than 90% CO2 capture combined with 97

Phycobilisome Mobility and Its Role in the Regulation of Light Harvesting in Red Algae

Czech Academy of Sciences Publication Activity Database

Kaňa, Radek; Kotabová, Eva; Lukeš, Martin; Papáček, Š.; Matonoha, Ctirad; Liu, L.N.; Prášil, Ondřej; Mullineaux, C.W.

2014-01-01

Roč. 165, č. 4 (2014), s. 1618-1631 ISSN 0032-0889 R&D Projects: GA ČR GAP501/12/0304; GA MŠk ED2.1.00/03.0110 Institutional support: RVO:61388971 ; RVO:67985807 Keywords : EXCITATION-ENERGY TRANSFER * CYANOBACTERIAL THYLAKOID MEMBRANE S * MICROALGA PORPHYRIDIUM-CRUENTUM Subject RIV: EE - Microbiology, Virology; BA - General Mathematics (UIVT-O) Impact factor: 6.841, year: 2014

Effect of magnetized extender on sperm membrane integrity and development of oocytes in vitro fertilized with liquid storage boar semen.

Science.gov (United States)

Lee, Sang-Hee; Park, Choon-Keun

2015-03-01

The objective of this study was to evaluate the effect of a magnetized extender on sperm membrane damage and development of oocytes in vitro fertilized with liquid storage boar semen. Before semen dilution, extender was flowed through a neodymium magnet (0, 2000, 4000 and 6000G) for 5min and collected semen was preserved for 168h at 18°C. In results, plasma membrane integrity with live sperm was significantly higher in semen treated with extenders magnetized at 4000G than sperm treated with extenders magnetized at 0G during semen preservation for 120-168h (psemen treated with extenders magnetized at 4000 and 6000G compared to 0 and 2000G during semen preservation for 168h (psemen treated with extenders magnetized at 2000G than other groups during semen preservation for 168h. The ability of semen to achieve successful in vitro fertilization was also not significantly different among the groups during preservation. However, when the semen was preserved for 168h, the blastocyst formation rates were significantly higher at 6000G compared to 0 and 2000G (psemen extender could protect the sperm membrane from damage, and improve the ability of rates of in vitro blastocyst development and magnetized semen diluter is beneficial for long liquid preservation of boar semen. Copyright © 2014 Elsevier B.V. All rights reserved.

Nanoengineered membranes for controlled transport

Science.gov (United States)

Doktycz, Mitchel J [Oak Ridge, TN; Simpson, Michael L [Knoxville, TN; McKnight, Timothy E [Greenback, TN; Melechko, Anatoli V [Oak Ridge, TN; Lowndes, Douglas H [Knoxville, TN; Guillorn, Michael A [Knoxville, TN; Merkulov, Vladimir I [Oak Ridge, TN

2010-01-05

A nanoengineered membrane for controlling material transport (e.g., molecular transport) is disclosed. The membrane includes a substrate, a cover definining a material transport channel between the substrate and the cover, and a plurality of fibers positioned in the channel and connected to an extending away from a surface of the substrate. The fibers are aligned perpendicular to the surface of the substrate, and have a width of 100 nanometers or less. The diffusion limits for material transport are controlled by the separation of the fibers. In one embodiment, chemical derivitization of carbon fibers may be undertaken to further affect the diffusion limits or affect selective permeability or facilitated transport. For example, a coating can be applied to at least a portion of the fibers. In another embodiment, individually addressable carbon nanofibers can be integrated with the membrane to provide an electrical driving force for material transport.

Comparison of membrane electroporation and protein denature in response to pulsed electric field with different durations.

Science.gov (United States)

Huang, Feiran; Fang, Zhihui; Mast, Jason; Chen, Wei

2013-05-01

In this paper, we compared the minimum potential differences in the electroporation of membrane lipid bilayers and the denaturation of membrane proteins in response to an intensive pulsed electric field with various pulse durations. Single skeletal muscle fibers were exposed to a pulsed external electric field. The field-induced changes in the membrane integrity (leakage current) and the Na channel currents were monitored to identify the minimum electric field needed to damage the membrane lipid bilayer and the membrane proteins, respectively. We found that in response to a relatively long pulsed electric shock (longer than the membrane intrinsic time constant), a lower membrane potential was needed to electroporate the cell membrane than for denaturing the membrane proteins, while for a short pulse a higher membrane potential was needed. In other words, phospholipid bilayers are more sensitive to the electric field than the membrane proteins for a long pulsed shock, while for a short pulse the proteins become more vulnerable. We can predict that for a short or ultrashort pulsed electric shock, the minimum membrane potential required to start to denature the protein functions in the cell plasma membrane is lower than that which starts to reduce the membrane integrity. Copyright © 2013 Wiley Periodicals, Inc.

Integration of sand and membrane filtration systems for iron and pesticide removal without chemical addition

DEFF Research Database (Denmark)

Kowalski, Krysztof; Madsen, Henrik Tækker; Søgaard, Erik Gydesen

2013-01-01

the content of key foulants, the techniques can be used as a pre-treatment for nanofiltration and low pressure reverse osmosis that has proved to be capable of removing pesticides. It was found that a lower fouling potential could be obtained by using the membranes, but that sand filter was better at removing......Pilot plant investigations of sand and membrane filtration (MF/UF/NF/LPRO) have been performed to treat groundwater polluted with pesticides. The results show that simple treatment, with use of aeration and sand filtration or MF/UF membranes, does not remove pesticides. However, by reducing...... manganese and dissolved organic matter. The results indicate that combining aeration; sand filtration and membrane techniques might be a good option for pesticide removal without any addition of chemicals and minimized membrane maintenance....

Biomimetic polymeric membranes for water treatment

DEFF Research Database (Denmark)

Habel, Joachim Erich Otto

This project is about the interplay of the three major components of aquaporin based biomimetic polymeric membranes (ABPMs): Aquaporins (AQPs), amphiphilic block copolymers, serving as a vesicular matrix for the hydrophobic AQP exterior (proteopolymersomes) and a polymeric membrane as embedment....... The interplay of proteopolymersomes and polymeric mesh support (in this case polyethersulfone, PES) was examined via integration of proteopolymersomes in an active layer (AL) formed by interfacial polymerisation between a linker molecule in aqueous phase and another in organic phase on top of the PES....... The resulting thin-film composite (TFC) membrane was analyzed via cross-flow forward osmosis (FO), scanning electron microscopy (SEM), fourier-transformed infrared spectroscopy (FTIR), as well as in the non-supported form over FTIR and a specialized microfluidic visualization approach. Where no clear dierences...

Characterization of mutants expressing thermostable D1 and D2 polypeptides of photosystem II in the cyanobacterium Synechococcus elongatus PCC 7942.

Science.gov (United States)

Haraguchi, Norihisa; Kaseda, Jun; Nakayama, Yasumune; Nagahama, Kazuhiro; Ogawa, Takahira; Matsuoka, Masayoshi

2018-06-08

Photosystem II complex embedded in thylakoid membrane performs oxygenic photosynthesis where the reaction center D1/D2 heterodimer accommodates all components of the electron transport chain. To express thermostable D1/D2 heterodimer in a cyanobacterium Synechococcus elongatus PCC 7942, we constructed a series of mutant strains whose psbA1 and psbD1 genes encoding, respectively, the most highly expressed D1 and D2 polypeptides were replaced with those of a thermophilic strain, Thermosynechococcus vulcanus. Because the C-terminal 16 amino acid sequences of D1 polypeptides should be processed prior to maturation but diverge from each other, we also constructed the psbA1ΔC-replaced strain expressing a thermostable D1 polypeptide devoid of the C-terminal extension. The psbA1/psbD1-replaced strain showed decreased growth rate and oxygen evolution rate, suggesting inefficient photosystem II. Immunoblot analyses for thermostable D1, D2 polypeptides revealed that the heterologous D1 protein was absent in thylakoid membrane from any mutant strains with psbA1, psbA1ΔC, and psbA1/psbD1-replacements, whereas the heterologous D2 protein was present in thylakoid membrane as well as purified photosystem II complex from the psbA1/psbD1-replaced strain. In the latter strain, the compensatory expression of psbA3 and psbD2 genes was elevated. These data suggest that heterologous D2 polypeptide could be combined with the host D1 polypeptide to form chimeric D1/D2 heterodimer, whereas heterologous D1 polypeptide even without the C-terminal extension was unable to make complex with the host D2 polypeptide. Since the heterologous D1 could not be detected even in the whole cells of psbA1/psbD1-replaced strain, the rapid degradation of unprocessed or unassembled heterologous D1 was implicated. Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Ceramic membrane ultrafiltration of natural surface water with ultrasound enhanced backwashing.

Science.gov (United States)

Boley, A; Narasimhan, K; Kieninger, M; Müller, W-R

2010-01-01

Ultrafiltration membrane cleaning with ultrasound enhanced backwashing was investigated with two ceramic membrane systems in parallel. One of them was subjected to ultrasound during backwashing, the other acted as a reference system. The feed water was directly taken from a creek with a sedimentation process as only pre-treatment. The cleaning performance was improved with ultrasound but after 3 weeks of operation damages occurred on the membranes. These effects were studied with online measurements of flux, trans-membrane-pressure and temperature, but also with integrity tests, turbidity measurements and visual examination.

NATO Advanced Study Institute on Synthetic Membranes : Science, Engineering and Applications

CERN Document Server

Lonsdale, H; Pinho, M

1986-01-01

The chapters in this book are based upon lectures given at the NATO Advanced Study Institute on Synthetic Membranes (June 26-July 8, 1983, Alcabideche, Portugal), which provided an integrated presentation of syn­ thetic membrane science and technology in three broad areas. Currently available membrane formation mechanisms are reviewed, as well as the manner in which synthesis conditions can be controlled to achieve desired membrane structures. Membrane performance in a specific separa­ tionprocess involves complex phenomena, the understanding of which re­ quires a multidisciplinary approach encompassing polymer chemistry, phys­ ical chemistry, and chemical engineering. Progress toward a global understanding of membrane phenomena is described in chapters on the principles of membrane transport. The chapters on membrane processes and applications highlight both established and emerging membrane processes, and elucidate their myriad applications. It is our hope that this book will be an enduring, comprehensi...

Modeling of an integrated fermentation/membrane extraction process for the production of 2-phenylethanol and 2-phenylethylacetate.

Science.gov (United States)

Adler, Philipp; Hugen, Thorsten; Wiewiora, Marzena; Kunz, Benno

2011-03-07

An unstructured model for an integrated fermentation/membrane extraction process for the production of the aroma compounds 2-phenylethanol and 2-phenylethylacetate by Kluyveromyces marxianus CBS 600 was developed. The extent to which this model, based only on data from the conventional fermentation and separation processes, provided an estimation of the integrated process was evaluated. The effect of product inhibition on specific growth rate and on biomass yield by both aroma compounds was approximated by multivariate regression. Simulations of the respective submodels for fermentation and the separation process matched well with experimental results. With respect to the in situ product removal (ISPR) process, the effect of reduced product inhibition due to product removal on specific growth rate and biomass yield was predicted adequately by the model simulations. Overall product yields were increased considerably in this process (4.0 g/L 2-PE+2-PEA vs. 1.4 g/L in conventional fermentation) and were even higher than predicted by the model. To describe the effect of product concentration on product formation itself, the model was extended using results from the conventional and the ISPR process, thus agreement between model and experimental data improved notably. Therefore, this model can be a useful tool for the development and optimization of an efficient integrated bioprocess. Copyright © 2010 Elsevier Inc. All rights reserved.

The ER in 3-D: a multifunctional dynamic membrane network

OpenAIRE

Friedman, Jonathan R.; Voeltz, Gia K.

2011-01-01

The endoplasmic reticulum (ER) is a large, singular, membrane-bound organelle that has an elaborate 3-D structure with a diversity of structural domains. It contains regions that are flat and cisternal, ones that are highly curved and tubular, and others adapted to form contact with nearly every other organelle and with the plasma membrane. ER 3-D structure is determined by both integral ER membrane proteins and by interactions with the cytoskeleton. Here, we describe some of the factors that...

Performance analysis of an integrated biomass gasification and PEMFC (proton exchange membrane fuel cell) system: Hydrogen and power generation

International Nuclear Information System (INIS)

Chutichai, Bhawasut; Authayanun, Suthida; Assabumrungrat, Suttichai; Arpornwichanop, Amornchai

2013-01-01

The PEMFC (proton exchange membrane fuel cell) is expected to play a significant role in next-generation energy systems. Because most hydrogen that is used as a fuel for PEMFCs is derived from the reforming of natural gas, the use of renewable energy sources such as biomass to produce this hydrogen offers a promising alternative. This study is focused on the performance analysis of an integrated biomass gasification and PEMFC system. The combined heat and power generation output of this integrated system is designed for residential applications, taking into account thermal and electrical demands. A flowsheet model of the integrated PEMFC system is developed and employed to analyze its performance with respect to various key operating parameters. A purification process consisting of a water–gas shift reactor and a preferential oxidation reactor is also necessary in order to reduce the concentration of CO in the synthesis gas to below 10 ppm for subsequent use in the PEMFC. The effect of load level on the performance of the PEMFC system is investigated. Based on an electrical load of 5 kW, it is found that the electrical efficiency of the PEMFC integrated system is 22%, and, when waste heat recovery is considered, the total efficiency of the PEMFC system is 51%. - Highlights: • Performance of a biomass gasification and PEMFC integrated system is analyzed. • A flowsheet model of the PEMFC integrated system is developed. • Effect of biomass sources and key parameters on hydrogen and power generation is presented. • The PEMFC integrated system is designed for small-scale power demand. • Effect of load changes on the performance of PEMFC is investigated

Novel Ultrafine Fibrous Poly(tetrafluoroethylene Hollow Fiber Membrane Fabricated by Electrospinning

Directory of Open Access Journals (Sweden)

Qinglin Huang

2018-04-01

Full Text Available Novel poly(tetrafluoroethylene (PTFE hollow fiber membranes were successfully fabricated by electrospinning, with ultrafine fibrous PTFE membranes as separation layers, while a porous glassfiber braided tube served as the supporting matrix. During this process, PTFE/poly(vinylalcohol (PVA ultrafine fibrous membranes were electrospun while covering the porous glassfiber braided tube; then, the nascent PTFE/PVA hollow fiber membrane was obtained. In the following sintering process, the spinning carrier PVA decomposed; meanwhile, the ultrafine fibrous PTFE membrane shrank inward so as to further integrate with the supporting matrix. Therefore, the ultrafine fibrous PTFE membranes had excellent interface bonding strength with the supporting matrix. Moreover, the obtained ultrafine fibrous PTFE hollow fiber membrane exhibited superior performances in terms of strong hydrophobicity (CA > 140°, high porosity (>70%, and sharp pore size distribution. The comprehensive properties indicated that the ultrafine fibrous PTFE hollow fiber membranes could have potentially useful applications in membrane contactors (MC, especially membrane distillation (MD in harsh water environments.

Electrospun polyurethane membranes for Tissue Engineering applications

Energy Technology Data Exchange (ETDEWEB)

Gabriel, Laís P., E-mail: lagabriel@gmail.com [National Institute of Biofabrication, Campinas (Brazil); Department of Chemical Engineering, University of Campinas, Campinas (Brazil); Rodrigues, Ana Amélia [National Institute of Biofabrication, Campinas (Brazil); Department of Medical Sciences, University of Campinas, Campinas (Brazil); Macedo, Milton; Jardini, André L.; Maciel Filho, Rubens [National Institute of Biofabrication, Campinas (Brazil); Department of Chemical Engineering, University of Campinas, Campinas (Brazil)

2017-03-01

Tissue Engineering proposes, among other things, tissue regeneration using scaffolds integrated with biological molecules, growth factors or cells for such regeneration. In this research, polyurethane membranes were prepared using the electrospinning technique in order to obtain membranes to be applied in Tissue Engineering, such as epithelial, drug delivery or cardiac applications. The influence of fibers on the structure and morphology of the membranes was studied using scanning electron microscopy (SEM), the structure was evaluated by Fourier transform infrared spectroscopy (FT-IR), and the thermal stability was analyzed by thermogravimetry analysis (TGA). In vitro cells attachment and proliferation was investigated by SEM, and in vitro cell viability was studied by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays and Live/Dead® assays. It was found that the membranes present an homogeneous morphology, high porosity, high surface area/volume ratio, it was also observed a random fiber network. The thermal analysis showed that the membrane degradation started at 254 °C. In vitro evaluation of fibroblasts cells showed that fibroblasts spread over the membrane surface after 24, 48 and 72 h of culture. This study supports the investigation of electrospun polyurethane membranes as biocompatible scaffolds for Tissue Engineering applications and provides some guidelines for improved biomaterials with desired properties.

Synthesis of zeolite NaA membrane from fused fly ash extract.

Science.gov (United States)

Ameh, Alechine E; Musyoka, Nicholas M; Fatoba, Ojo O; Syrtsova, Daria A; Teplyakov, Vladimir V; Petrik, Leslie F

2016-01-01

Zeolite-NaA membranes were synthesized from an extract of fused South African fly ash on a porous titanium support by a secondary growth method. The influence of the synthesis molar regime on the formation of zeolite NaA membrane layer was investigated. Two synthesis mixtures were generated by adding either aluminium hydroxide or sodium aluminate to the fused fly ash extract. The feedstock material and the synthesized membranes were characterized by X-diffraction (XRD), scanning electron microscopy (SEM) and X-ray fluorescence spectroscopy (XRF). It was found by XRD and SEM that the cubic crystals of a typical zeolite NaA with a dense intergrown layer was formed on the porous Ti support. The study shows that the source of Al used had an effect on the membrane integrity as sodium aluminate provided the appropriate amount of Na(+) to form a coherent membrane of zeolite NaA, whereas aluminium hydroxide did not. Morphological, the single hydrothermal stage seeded support formed an interlocked array of zeolite NaA particles with neighbouring crystals. Also, a robust, continuous and well-intergrown zeolite NaA membrane was formed with neighbouring crystals of zeolite fused to each other after the multiple stage synthesis. The synthesized membrane was permeable to He (6.0 × 10(6) L m(-2)h(-1) atm(-1)) and CO2 (5.6 × 10(6) L m(-2)h(-1) atm(-1)), which indicate that the layer of the membrane was firmly attached to the porous Ti support. Membrane selectivity was maintained showing membrane integrity with permselectivity of 1.1, showing that a waste feedstock, fly ash, could be utilized for preparing robust zeolite NaA membranes on Ti support.

Membranes for Redox Flow Battery Applications