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  • Articles  (65)
  • Biotechnology & Synthetic Biology  (41)
  • Ocean circulation  (24)
  • Oxford University Press  (41)
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  • 2015-2019  (65)
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  • 1
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Climate 28 (2015): 8574–8584, doi:10.1175/JCLI-D-14-00809.1.
    Description: The subsurface ocean response to anthropogenic climate forcing remains poorly characterized. From the Coupled Model Intercomparison Project (CMIP), a robust response of the lower thermocline is identified, where the warming is considerably weaker in the subtropics than in the tropics and high latitudes. The lower thermocline change is inversely proportional to the thermocline depth in the present climatology. Ocean general circulation model (OGCM) experiments show that sea surface warming is the dominant forcing for the subtropical gyre change in contrast to natural variability for which wind dominates, and the ocean response is insensitive to the spatial pattern of surface warming. An analysis based on a ventilated thermocline model shows that the pattern of the lower thermocline change can be interpreted in terms of the dynamic response to the strengthened stratification and downward heat mixing. Consequently, the subtropical gyres become intensified at the surface but weakened in the lower thermcline, consistent with results from CMIP experiments.
    Description: The work was supported by the National Basic Research Program of China (2012CB955600), the National Natural Science Foundation of China (41125019, 41206021), and the U.S. National Science Foundation (AGS 1249145, 1305719).
    Description: 2016-05-01
    Keywords: Circulation/ Dynamics ; Ocean circulation ; Physical Meteorology and Climatology ; Climate change
    Repository Name: Woods Hole Open Access Server
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  • 2
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 2127-2140, doi:10.1175/JPO-D-18-0035.1.
    Description: Shipboard hydrographic and velocity measurements collected in summer 2014 are used to study the evolution of the freshwater coastal current in southern Greenland as it encounters Cape Farewell. The velocity structure reveals that the coastal current maintains its identity as it flows around the cape and bifurcates such that most of the flow is diverted to the outer west Greenland shelf, while a small portion remains on the inner shelf. Taking into account this inner branch, the volume transport of the coastal current is conserved, but the freshwater transport decreases on the west side of Cape Farewell. A significant amount of freshwater appears to be transported off the shelf where the outer branch flows adjacent to the shelfbreak circulation. It is argued that the offshore transposition of the coastal current is caused by the flow following the isobaths as they bend offshore because of the widening of the shelf on the west side of Cape Farewell. An analysis of the potential vorticity shows that the subsequent seaward flux of freshwater can be enhanced by instabilities of the current. This set of circumstances provides a pathway for the freshest water originating from the Arctic, as well as runoff from the Greenland ice sheet, to be fluxed into the interior Labrador Sea where it could influence convection in the basin.
    Description: Funding for this project was provided by the National Science Foundation under Grant OCE-1259618.
    Description: 2019-03-11
    Keywords: Boundary currents ; Coastal flows ; Instability ; Ocean circulation ; Potential vorticity ; Transport
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  • 3
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 966–987, doi:10.1175/JPO-D-14-0110.1.
    Description: A key remaining challenge in oceanography is the understanding and parameterization of small-scale mixing. Evidence suggests that topographic features play a significant role in enhancing mixing in the Southern Ocean. This study uses 914 high-resolution hydrographic profiles from novel EM-APEX profiling floats to investigate turbulent mixing north of the Kerguelen Plateau, a major topographic feature in the Southern Ocean. A shear–strain finescale parameterization is applied to estimate diapycnal diffusivity in the upper 1600 m of the ocean. The indirect estimates of mixing match direct microstructure profiler observations made simultaneously. It is found that mixing intensities have strong spatial and temporal variability, ranging from O(10−6) to O(10−3) m2 s−1. This study identifies topographic roughness, current speed, and wind speed as the main factors controlling mixing intensity. Additionally, the authors find strong regional variability in mixing dynamics and enhanced mixing in the Antarctic Circumpolar Current frontal region. This enhanced mixing is attributed to dissipating internal waves generated by the interaction of the Antarctic Circumpolar Current and the topography of the Kerguelen Plateau. Extending the mixing observations from the Kerguelen region to the entire Southern Ocean, this study infers a large water mass transformation rate of 17 Sverdrups (Sv; 1 Sv ≡ 106 m3 s−1) across the boundary of Antarctic Intermediate Water and Upper Circumpolar Deep Water in the Antarctic Circumpolar Current. This work suggests that the contribution of mixing to the Southern Ocean overturning circulation budget is particularly significant in fronts.
    Description: AM was supported by the joint CSIRO–University of Tasmania Quantitative Marine Science (QMS) program and the 2009 CSIRO Wealth from Ocean Flagship Collaborative Fund. BMS was supported by the Australian Climate Change Science Program, jointly funded by the Department of the Environment and CSIRO. KLPs salary support was provided by Woods Hole Oceanographic Institution bridge support funds.
    Description: 2015-10-01
    Keywords: Geographic location/entity ; Southern Ocean ; Circulation/ Dynamics ; Diapycnal mixing ; Fronts ; Ocean circulation ; Topographic effects ; Atm/Ocean Structure/ Phenomena ; Mixing
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  • 4
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 2598–2620, doi:10.1175/JPO-D-14-0249.1.
    Description: Through combining analytical arguments and numerical models, this study investigates the finite-amplitude meanders of shelfbreak fronts characterized by sloping isopycnals outcropping at both the surface and the shelfbreak bottom. The objective is to provide a formula for the meander length scale that can explain observed frontal length scale variability and also be verified with observations. Considering the frontal instability to be a mixture of barotropic and baroclinic instability, the derived along-shelf meander length scale formula is [b1/(1 + a1S1/2)]NH/f, where N is the buoyancy frequency; H is the depth of the front; f is the Coriolis parameter; S is the Burger number measuring the ratio of energy conversion associated with barotropic and baroclinic instability; and a1 and b1 are empirical constants. Initial growth rate of the frontal instability is formulated as [b2(1 + a1S1/2)/(1 + a2αS1/2)]NH/L, where α is the bottom slope at the foot of the front, and a2 and b2 are empirical constants. The formulas are verified using numerical sensitivity simulations, and fitting of the simulated and formulated results gives a1 = 2.69, b1 = 14.65, a2 = 5.1 × 103, and b2 = 6.2 × 10−2. The numerical simulations also show development of fast-growing frontal symmetric instability when the minimum initial potential vorticity is negative. Although frontal symmetric instability leads to faster development of barotropic and baroclinic instability at later times, it does not significantly influence the meander length scale. The derived meander length scale provides a framework for future studies of the influences of external forces on shelfbreak frontal circulation and cross-frontal exchange.
    Description: WGZ and GGG were supported by the National Science Foundation through Grant OCE-1129125.
    Description: 2016-04-01
    Keywords: Circulation/ Dynamics ; Instability ; Ocean circulation ; Topographic effects ; Atm/Ocean Structure/ Phenomena ; Fronts ; Models and modeling ; Numerical analysis/modeling
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  • 5
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 2913–2932, doi:10.1175/JPO-D-14-0179.1.
    Description: The oceanic deep circulation is shared between concentrated deep western boundary currents (DWBCs) and broader interior pathways, a process that is sensitive to seafloor topography. This study investigates the spreading and deepening of Denmark Strait overflow water (DSOW) in the western subpolar North Atlantic using two ° eddy-resolving Atlantic simulations, including a passive tracer injected into the DSOW. The deepest layers of DSOW transit from a narrow DWBC in the southern Irminger Sea into widespread westward flow across the central Labrador Sea, which remerges along the Labrador coast. This abyssal circulation, in contrast to the upper levels of overflow water that remain as a boundary current, blankets the deep Labrador Sea with DSOW. Farther downstream after being steered around the abrupt topography of Orphan Knoll, DSOW again leaves the boundary, forming cyclonic recirculation cells in the deep Newfoundland basin. The deep recirculation, mostly driven by the meandering pathway of the upper North Atlantic Current, leads to accumulation of tracer offshore of Orphan Knoll, precisely where a local maximum of chlorofluorocarbon (CFC) inventory is observed. At Flemish Cap, eddy fluxes carry ~20% of the tracer transport from the boundary current into the interior. Potential vorticity is conserved as the flow of DSOW broadens at the transition from steep to less steep continental rise into the Labrador Sea, while around the abrupt topography of Orphan Knoll, potential vorticity is not conserved and the DSOW deepens significantly.
    Description: This work is supported by ONR Award N00014-09-1-0587, the NSF Physical Oceanography Program, and NASA Ocean Surface Topography Science Team Program.
    Description: 2016-06-01
    Keywords: Circulation/ Dynamics ; Abyssal circulation ; Boundary currents ; Ocean circulation ; Ocean dynamics ; Potential vorticity ; Topographic effects
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  • 6
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2016. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 46 (2016): 3011-3029, doi:10.1175/JPO-D-15-0248.1.
    Description: Seasonal variability of the tropical Atlantic circulation is dominated by the annual cycle, but semiannual variability is also pronounced, despite weak forcing at that period. This study uses multiyear, full-depth velocity measurements from the central equatorial Atlantic to analyze the vertical structure of annual and semiannual variations of zonal velocity. A baroclinic modal decomposition finds that the annual cycle is dominated by the fourth mode and the semiannual cycle is dominated by the second mode. Similar local behavior is found in a high-resolution general circulation model. This simulation reveals that the annual and semiannual cycles of the respective dominant baroclinic modes are associated with characteristic basinwide structures. Using an idealized, linear, reduced-gravity model to simulate the dynamics of individual baroclinic modes, it is shown that the observed circulation variability can be explained by resonant equatorial basin modes. Corollary simulations of the reduced-gravity model with varying basin geometry (i.e., square basin vs realistic coastlines) or forcing (i.e., spatially uniform vs spatially variable wind) show a structural robustness of the simulated basin modes. A main focus of this study is the seasonal variability of the Equatorial Undercurrent (EUC) as identified in recent observational studies. Main characteristics of the observed EUC including seasonal variability of transport, core depth, and maximum core velocity can be explained by the linear superposition of the dominant equatorial basin modes as obtained from the reduced-gravity model.
    Description: This study was supported by the Deutsche Forschungsgemeinschaft as part of the Sonderforschungsbereich 754 (SFB754) ‘‘Climate–Biogeochemistry Interactions in the Tropical Ocean’’ and through several research cruises with R/V Meteor, R/V Maria S. Merian, andR/VL’Atalante by the German Federal Ministry of Education and Research as part of the cooperative projects RACE (03F0605B) and SACUS (03G0837A) and by European Union 7th Framework Programme (FP7 2007–13) under Grant Agreement 603521 PREFACE project.
    Keywords: Atlantic Ocean ; Ocean circulation ; In situ oceanic observations ; Ocean models ; Seasonal cycle ; Tropical variability
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  • 7
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2017. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 47 (2017): 339-351, doi:10.1175/JPO-D-16-0165.1.
    Description: A novel multi-iteration statistical method for studying tracer spreading using drifter data is introduced. The approach allows for the best use of the available drifter data by making use of a simple iterative procedure, which results in the statistically probable map showing the likelihood that a tracer released at some source location would visit different geographical regions, along with the associated arrival travel times. The technique is tested using real drifter data in the North Atlantic. Two examples are considered corresponding to sources in the western and eastern North Atlantic Ocean, that is, Massachusetts Bay–like and Irish Sea–like sources, respectively. In both examples, the method worked well in estimating the statistics of the tracer transport pathways and travel times throughout the entire North Atlantic. The role of eddies versus mean flow is quantified using the same technique, and eddies are shown to significantly broaden the spread of a tracer. The sensitivity of the results to the size of the source domain is investigated and causes for this sensitivity are discussed.
    Description: This work was supported by the Grant OCE-1356630 from the National Science Foundation (NSF). Rypina also acknowledges NSF Grant OCE-1154641 and NASA Grant NNX14AH29G.
    Description: 2017-07-31
    Keywords: Atlantic Ocean ; Mass fluxes/transport ; Ocean circulation ; Trajectories ; Statistics
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  • 8
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2017. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 47 (2017): 633-647, doi:10.1175/JPO-D-16-0089.1.
    Description: Interannual variability in the volumetric water mass distribution within the North Atlantic Subtropical Gyre is described in relation to variability in the Atlantic meridional overturning circulation. The relative roles of diabatic and adiabatic processes in the volume and heat budgets of the subtropical gyre are investigated by projecting data into temperature coordinates as volumes of water using an Argo-based climatology and an ocean state estimate (ECCO version 4). This highlights that variations in the subtropical gyre volume budget are predominantly set by transport divergence in the gyre. A strong correlation between the volume anomaly due to transport divergence and the variability of both thermocline depth and Ekman pumping over the gyre suggests that wind-driven heave drives transport anomalies at the gyre boundaries. This wind-driven heaving contributes significantly to variations in the heat content of the gyre, as do anomalies in the air–sea fluxes. The analysis presented suggests that wind forcing plays an important role in driving interannual variability in the Atlantic meridional overturning circulation and that this variability can be unraveled from spatially distributed hydrographic observations using the framework presented here.
    Description: DGE was supported by a Natural Environment Research Council studentship award at the University of Southampton. JMT’s contribution was supported by the U.S. National Science Foundation (Grant OCE-1332667). GF’s contribution was supported by the U.S. National Science Foundation through Grant OCE-0961713 and by the U.S. National Oceanic and Atmospheric Administration through Grant NA10OAR4310135. The contributions of JDZ and AJGN were supported by the NERC Grant ‘‘Climate scale analysis of air and water masses’’ (NE/ K012932/1). ACNG gratefully acknowledges support from the Leverhulme Trust, the Royal Society, and the Wolfson Foundation. LY was supported by NASA Ocean Vector Wind Science Team (OVWST) activities under Grant NNA10AO86G.
    Keywords: North Atlantic Ocean ; Atmosphere-ocean interaction ; Ekman pumping/transport ; Ocean circulation ; Water masses ; Inverse methods
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  • 9
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 573-590, doi:10.1175/JPO-D-17-0206.1.
    Description: Motivated by the proximity of the Northern Recirculation Gyre and the deep western boundary current in the North Atlantic, an idealized model is used to investigate how recirculation gyres and a deep flow along a topographic slope interact. In this two-layer quasigeostrophic model, an unstable jet imposed in the upper layer generates barotropic recirculation gyres. These are maintained by an eddy-mean balance of potential vorticity (PV) in steady state. The authors show that the topographic slope can constrain the northern recirculation gyre meridionally and that the gyre’s adjustment to the slope leads to increased eddy PV fluxes at the base of the slope. When a deep current is present along the topographic slope in the lower layer, these eddy PV fluxes stir the deep current and recirculation gyre waters. Increased proximity to the slope dampens the eddy growth rate within the unstable jet, altering the geometry of recirculation gyre forcing and leading to a decrease in overall eddy PV fluxes. These mechanisms may shape the circulation in the western North Atlantic, with potential feedbacks on the climate system.
    Description: We gratefully acknowledge an AMS graduate fellowship (IALB) and U.S. National Science Foundation Grants OCE-1332667 and 1332834 (IALB and JMT).
    Description: 2018-09-06
    Keywords: Boundary currents ; Meridional overturning circulation ; Mesoscale processes ; Ocean circulation ; Potential vorticity ; Quasigeostrophic models
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  • 10
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 739-748, doi:10.1175/JPO-D-17-0089.1.
    Description: McDougall and Ferrari have estimated the global deep upward diapycnal flow in the boundary layer overlying continental slopes that must balance both downward diapycnal flow in the deep interior and the formation of bottom water around Antarctica. The decrease of perimeter of isopycnal surfaces with depth and the observed decay with height above bottom of turbulent dissipation in the deep ocean play a key role in their estimate. They argue that because the perimeter of seamounts increases with depth, the net effect of mixing around seamounts is to produce net downward diapycnal flow. While this is true along much of a seamount, it is shown here that diapycnal flow of the densest water around the seamount is upward, with buoyancy being transferred from water just above. The same is true for midocean ridges, whose perimeter is constant with depth. It is argued that mixing around seamounts and especially midocean ridges contributes positively to the global deep overturning circulation, reducing the amount of turbulence demanded over the continental slopes to balance the buoyancy budget for the bottom and deep water.
    Description: This work was supported by National Science Foundation Grant OCE- 1232962.
    Description: 2018-09-29
    Keywords: Abyssal circulation ; Boundary currents ; Buoyancy ; Diapycnal mixing ; Mass fluxes/transport ; Ocean circulation
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  • 11
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    American Meteorological Society
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 643-646, doi:10.1175/JPO-D-17-0240.1.
    Description: A simple oceanic model is presented for source–sink flow on the β plane to discuss the pathways from source to sink when transport boundary layers have large enough Reynolds numbers to be inertial in their dynamics. A representation of the flow as a Fofonoff gyre, suggested by prior work on inertial boundary layers and eddy-driven circulations in two-dimensional turbulent flows, indicates that even when the source and sink are aligned along the same western boundary the flow must intrude deep into the interior before exiting at the sink. The existence of interior pathways for the flow is thus an intrinsic property of an inertial circulation and is not dependent on particular geographical basin geometry.
    Description: 2018-09-12
    Keywords: Abyssal circulation ; Bottom currents ; Nonlinear dynamics ; Ocean circulation ; Ocean dynamics ; Potential vorticity
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  • 12
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    American Meteorological Society
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 1831-1848, doi:10.1175/JPO-D-18-0068.1.
    Description: We present a simplified theory using reduced-gravity equations for North Atlantic Deep Water (NADW) and its variation driven by high-latitude deep-water formation. The theory approximates layer thickness on the eastern boundary with domain-averaged layer thickness and, in tandem with a mass conservation argument, retains fundamental physics for cross-equatorial flows on interannual and longer forcing time scales. Layer thickness anomalies are driven by a time-dependent northern boundary condition that imposes a southward volume flux representative of a variable source of NADW and damped by diapycnal mixing throughout the basin. Moreover, an outflowing southern boundary condition imposes a southward volume flux that generally differs from the volume flux at the northern boundary, giving rise to temporal storage of NADW within the Atlantic basin. Closed form analytic solutions for the amplitude and phase are provided when the variable source of NADW is sinusoidal. We provide a nondimensional analysis that demonstrates that solution behavior is primarily controlled by two parameters that characterize the meridional extent of the southern basin and the width of the basin relative to the equatorial deformation radius. Similar scaling applied to the time-lagged equations of Johnson and Marshall provides a clear connection to their results. Numerical simulations of reduced-gravity equations agree with analytic predictions in linear, turbulent, and diabatic regimes. The theory introduces a simple analytic framework for studying idealized buoyancy- and wind-driven cross-equatorial flows on interannual and longer time scales.
    Description: This research was supported by the National Science Foundation under Grant OCE- 1634468.
    Description: 2019-02-15
    Keywords: North Atlantic Ocean ; Tropics ; Meridional overturning circulation ; Ocean circulation ; Shallow-water equations
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  • 13
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 48 (2018): 883-904, doi:10.1175/JPO-D-17-0084.1.
    Description: The dynamics controlling the along-valley (cross shelf) flow in idealized shallow shelf valleys with small to moderate Burger number are investigated, and analytical scales of the along-valley flows are derived. This paper follows Part I, which shows that along-shelf winds in the opposite direction to coastal-trapped wave propagation (upwelling regime) force a strong up-valley flow caused by the formation of a lee wave. In contrast, along-shelf winds in the other direction (downwelling regime) do not generate a lee wave and consequently force a relatively weak net down-valley flow. The valley flows in both regimes are cyclostrophic with 0(1) Rossby number. A major difference between the two regimes is the along-shelf length scales of the along-valley flows L. In the upwelling regime Ls, depends on the valley width W, and the wavelength lambda(1w) of the coastal-trapped lee wave arrested by the along-shelf flow U-s. In the downwelling regime L depends on the inertial length scale U-s|'f and W-c. The along-valley velocity scale in the upwelling regime, given by V-u approximate to root pi H-c/H-s integral W-c lambda(1w)/2 pi L-x (1+L-x(2)/L-c(2))(-1) e(-(pi Wc)/(lambda 1w),) is based on potential vorticity (PV) conservation and lee-wave dynamics (Hs and H, are the shelf and valley depth scales, respectively, and fis the Coriolis parameter). The velocity scale in the downwelling regime, given by |v(d)| approximate to (H-s/H-s)[1 + (L-x(2)/L-x(2))](-1) fL, is based on PV conservation. The velocity scales are validated by the numerical sensitivity simulations and can be useful for observational studies of along -valley transports. The work provides a framework for investigating cross -shelf transport induced by irregular shelf bathymetry and calls for future studies of this type under realistic environmental conditions and over a broader parameter space.
    Description: Both WGZ and SJL were supported by the National Science Foundation (NSF) through Grant OCE 1154575.WGZis also supported by the NSF Grant OCE 1634965 and SJL by NSF Grant OCE 1558874.
    Description: 2018-10-16
    Keywords: Ocean circulation ; Topographic effects ; Upwelling/downwelling ; Waves, oceanic ; Wind stress ; Ocean models
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  • 14
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015):1189–1204, doi:10.1175/JPO-D-14-0122.1.
    Description: Winter outcropping of the Eighteen Degree Water (EDW) and its subsequent dispersion are studied using a ° eddy-resolving simulation of the Family of Linked Atlantic Modeling Experiments (FLAME). Outcropped EDW columns in the model simulations are detected in each winter from 1990 to 1999, and particles are deployed in the center of each outcropped EDW column. Subsequently, the trajectories of these particles are calculated for the following 5 yr. The particles slowly spread away from the outcropping region into the nonoutcropping/subducted EDW region south of ~30°N and eventually to the non-EDW region in the greater subtropical gyre. Approximately 30% of the particles are found in non-EDW waters 1 yr after deployment; after 5 yr, only 25% of the particles are found within EDW. The reoutcropping time is defined as the number of years between when a particle is originally deployed in an outcropping EDW column and when that particle is next found in an outcropping EDW column. Of the particles, 66% are found to reoutcrop as EDW in 1 yr, and less than 5% of the particles outcrop in each of the subsequent 4 yr. While the individual trajectories exhibit significant eddy-like motions, the time scale of reoutcropping is primarily set by the mean circulation. The dominance of reoutcropping in 1 yr suggests that EDW outcropping contributes considerably to the persistence of surface temperature anomalies from one winter to the next, that is, the reemergence of winter sea surface temperature anomalies.
    Description: We gratefully acknowledge the support from the NSF OCE Physical Oceanography program (NSF OCE-0961090 to Y-OK and J-JP; NSF OCE-0960776 to MSL and SFG; and NSF OCE-1242989 to Y-OK).
    Description: 2015-10-01
    Keywords: Circulation/ Dynamics ; Ocean circulation ; Atm/Ocean Structure/ Phenomena ; Water masses
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  • 15
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 1410–1425, doi:10.1175/JPO-D-14-0192.1.
    Description: The west-to-east crossover of boundary currents has been seen in mean circulation schemes from several past models of the Red Sea. This study investigates the mechanisms that produce and control the crossover in an idealized, eddy-resolving numerical model of the Red Sea. The authors also review the observational evidence and derive an analytical estimate for the crossover latitude. The surface buoyancy loss increases northward in the idealized model, and the resultant mean circulation consists of an anticyclonic gyre in the south and a cyclonic gyre in the north. In the midbasin, the northward surface flow crosses from the western boundary to the eastern boundary. Numerical experiments with different parameters indicate that the crossover latitude of the boundary currents changes with f0, β, and the meridional gradient of surface buoyancy forcing. In the analytical estimate, which is based on quasigeostrophic, β-plane dynamics, the crossover is predicted to lie at the latitude where the net potential vorticity advection (including an eddy component) is zero. Various terms in the potential vorticity budget can be estimated using a buoyancy budget, a thermal wind balance, and a parameterization of baroclinic instability.
    Description: This work is supported by Award USA 00002, KSA 00011, and KSA 00011/02 made by King Abdullah University of Science and Technology (KAUST), by National Science Foundation Grants OCE0927017, OCE1154641, and OCE85464100, and by the Woods Hole Oceanographic Institution Academic Program Office.
    Description: 2015-11-01
    Keywords: Circulation/ Dynamics ; Boundary currents ; Buoyancy ; Ocean circulation ; Ocean dynamics
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  • 16
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 2806–2819, doi:10.1175/JPO-D-15-0061.1.
    Description: An eastward-flowing current of a homogeneous fluid with velocity U, contained in a channel of width L, impinges on an island of width of O(L), and the resulting interaction and dynamics are studied for values of the supercriticality parameter, b = βL2/U, both larger and smaller than π2. The former case is subcritical with respect to Rossby waves, and the latter is supercritical. The nature of the flow field depends strongly on b, and in particular, the nature of the flow around the island and the proportion of the flow passing to the north or south of the island are sensitive to b and to the position of the island in the channel. The problem is studied analytically in a relatively simple, nonlinear quasigeostrophic and adiabatic framework and numerically with a shallow-water model that allows a qualitative extension of the results to the equator. Although the issues involved are motivated by the interaction of the Equatorial Undercurrent and the Galapagos Islands, the analysis presented here focuses on the fundamental issue of the distinctive nature of the flow as a function of Rossby wave criticality.
    Description: Supported by the National Science Foundation Grant OCE-0959381.
    Description: 2016-05-01
    Keywords: Circulation/ Dynamics ; Ocean circulation ; Ocean dynamics ; Waves, oceanic
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  • 17
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2014. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 44 (2014): 3033–3053, doi:10.1175/JPO-D-13-0227.1.
    Description: The East Greenland Current (EGC) had long been considered the main pathway for the Denmark Strait overflow (DSO). Recent observations, however, indicate that the north Icelandic jet (NIJ), which flows westward along the north coast of Iceland, is a major separate pathway for the DSO. In this study a two-layer numerical model and complementary integral constraints are used to examine various pathways that lead to the DSO and to explore plausible mechanisms for the NIJ’s existence. In these simulations, a westward and NIJ-like current emerges as a robust feature and a main pathway for the Denmark Strait overflow. Its existence can be explained through circulation integrals around advantageous contours. One such constraint spells out the consequences of overflow water as a source of low potential vorticity. A stronger constraint can be added when the outflow occurs through two outlets: it takes the form of a circulation integral around the Iceland–Faroe Ridge. In either case, the direction of overall circulation about the contour can be deduced from the required frictional torques. Some effects of wind stress forcing are also examined. The overall positive curl of the wind forces cyclonic gyres in both layers, enhancing the East Greenland Current. The wind stress forcing weakens but does not eliminate the NIJ. It also modifies the sign of the deep circulation in various subbasins and alters the path by which overflow water is brought to the Faroe Bank Channel, all in ways that bring the idealized model more in line with observations. The sequence of numerical experiments separates the effects of wind and buoyancy forcing and shows how each is important.
    Description: This study has been supported by National Science Foundation (OCE0927017 and ARC1107412).
    Description: 2015-06-01
    Keywords: Circulation/ Dynamics ; Boundary currents ; Channel flows ; Meridional overturning circulation ; Ocean circulation ; Topographic effects
    Repository Name: Woods Hole Open Access Server
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  • 18
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2016. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 46 (2016): 3263-3278, doi:10.1175/JPO-D-16-0091.1.
    Description: The halocline of the Beaufort Gyre varies significantly on interannual to decadal time scales, affecting the freshwater content (FWC) of the Arctic Ocean. This study explores the role of eddies in the Ekman-driven gyre variability. Following the transformed Eulerian-mean paradigm, the authors develop a theory that links the FWC variability to the stability of the large-scale gyre, defined as the inverse of its equilibration time. The theory, verified with eddy-resolving numerical simulations, demonstrates that the gyre stability is explicitly controlled by the mesoscale eddy diffusivity. An accurate representation of the halocline dynamics requires the eddy diffusivity of 300 ± 200 m2 s−1, which is lower than what is used in most low-resolution climate models. In particular, on interannual and longer time scales the eddy fluxes and the Ekman pumping provide equally important contributions to the FWC variability. However, only large-scale Ekman pumping patterns can significantly alter the FWC, with spatially localized perturbations being an order of magnitude less efficient. Lastly, the authors introduce a novel FWC tendency diagnostic—the Gyre Index—that can be conveniently calculated using observations located only along the gyre boundaries. Its strong predictive capabilities, assessed in the eddy-resolving model forced by stochastic winds, suggest that the Gyre Index would be of use in interpreting FWC evolution in observations as well as in numerical models.
    Description: GEMacknowledges the support from theHowland Postdoctoral Program Fund at WHOI and the Stanback Fellowship Fund at Caltech.MAS was supported by NSF Grants PLR-1415489 and OCE-1232389. AFT acknowledges support from NASA Award NNN12AA01C.
    Description: 2017-04-20
    Keywords: Arctic ; Eddies ; Ekman pumping/transport ; Large-scale motions ; Ocean circulation ; Stability
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  • 19
    Publication Date: 2022-05-25
    Description: Author Posting. © American Meteorological Society, 2018. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Climate 31 (2018): 9881-9901, doi:10.1175/JCLI-D-17-0889.1.
    Description: The Atlantic meridional overturning circulation and associated poleward heat transport are balanced by northern heat loss to the atmosphere and corresponding water-mass transformation. The circulation of northward-flowing Atlantic Water at the surface and returning overflow water at depth is particularly manifested—and observed—at the Greenland–Scotland Ridge where the water masses are guided through narrow straits. There is, however, a rich variability in the exchange of water masses across the ridge on all time scales. Focusing on seasonal and interannual time scales, and particularly the gateways of the Denmark Strait and between the Faroe Islands and Shetland, we specifically assess to what extent the exchanges of water masses across the Greenland–Scotland Ridge relate to wind forcing. On seasonal time scales, the variance explained of the observed exchanges can largely be related to large-scale wind patterns, and a conceptual model shows how this wind forcing can manifest via a barotropic, cyclonic circulation. On interannual time scales, the wind stress impact is less direct as baroclinic mechanisms gain importance and observations indicate a shift in the overflows from being more barotropically to more baroclinically forced during the observation period. Overall, the observed Greenland–Scotland Ridge exchanges reflect a horizontal (cyclonic) circulation on seasonal time scales, while the interannual variability more represents an overturning circulation.
    Description: This research was supported by the Research Council of Norway project NORTH (Grant 229763). Additional support for M. A. Spall was provided by National Science Foundation Grant OCE- 1558742, for T. Eldevik and S. Østerhus by the European Union’s Horizon 2020 research and innovation program project Blue-Action (Grant 727852), and for S. Østerhus by the European Framework Programs under Grant Agreement 308299 (NACLIM).
    Keywords: Ocean circulation ; Thermocline circulation ; Atmosphere-ocean interaction ; North Atlantic Oscillation ; Statistical techniques ; Time series
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  • 20
    Publication Date: 2022-05-26
    Description: Author Posting. © American Meteorological Society, 2015. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 45 (2015): 778–791, doi:10.1175/JPO-D-14-0164.1.
    Description: This study examines anisotropic transport properties of the eddying North Atlantic flow, using an idealized model of the double-gyre oceanic circulation and altimetry-derived velocities. The material transport by the time-dependent flow (quantified by the eddy diffusivity tensor) varies geographically and is anisotropic, that is, it has a well-defined direction of the maximum transport. One component of the time-dependent flow, zonally elongated large-scale transients, is particularly important for the anisotropy, as it corresponds to primarily zonal material transport and long correlation time scales. The importance of these large-scale zonal transients in the material distribution is further confirmed with simulations of idealized color dye tracers, which has implications for parameterizations of the eddy transport in non-eddy-resolving models.
    Description: IK would like to acknowledge support through the NSF Grant OCE-1154923. IR was supported by the NSF OCE-1154641 and NASA Grant NNX14AH29G.
    Description: 2015-09-01
    Keywords: Circulation/ Dynamics ; Eddies ; Lagrangian circulation/transport ; Mesoscale processes ; Ocean circulation ; Models and modeling ; Tracers
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  • 21
    Publication Date: 2022-05-26
    Description: © The Author(s), 2019. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Semper, S., Vage, K., Pickart, R. S., Valdimarsson, H., Torres, D. J., & Jonsson, S. The emergence of the North Icelandic Jet and its evolution from northeast Iceland to Denmark Strait. Journal of Physical Oceanography, 49(10), (2019): 2499-2521, doi:10.1175/JPO-D-19-0088.1.
    Description: The North Icelandic Jet (NIJ) is an important source of dense water to the overflow plume passing through Denmark Strait. The properties, structure, and transport of the NIJ are investigated for the first time along its entire pathway following the continental slope north of Iceland, using 13 hydrographic/velocity surveys of high spatial resolution conducted between 2004 and 2018. The comprehensive dataset reveals that the current originates northeast of Iceland and increases in volume transport by roughly 0.4 Sv (1 Sv ≡ 106 m3 s−1) per 100 km until 300 km upstream of Denmark Strait, at which point the highest transport is reached. The bulk of the NIJ transport is confined to a small area in Θ–S space centered near −0.29° ± 0.16°C in Conservative Temperature and 35.075 ± 0.006 g kg−1 in Absolute Salinity. While the hydrographic properties of this transport mode are not significantly modified along the NIJ’s pathway, the transport estimates vary considerably between and within the surveys. Neither a clear seasonal signal nor a consistent link to atmospheric forcing was found, but barotropic and/or baroclinic instability is likely active in the current. The NIJ displays a double-core structure in roughly 50% of the occupations, with the two cores centered at the 600- and 800-m isobaths, respectively. The transport of overflow water 300 km upstream of Denmark Strait exceeds 1.8 ± 0.3 Sv, which is substantially larger than estimates from a year-long mooring array and hydrographic/velocity surveys closer to the strait, where the NIJ merges with the separated East Greenland Current. This implies a more substantial contribution of the NIJ to the Denmark Strait overflow plume than previously envisaged.
    Description: Six different research vessels were involved in the collection of the data used in this study: RRS James Clark Ross, R/V Knorr, R/V Bjarni Sæmundsson, R/V Håkon Mosby, NRV Alliance, and R/V Kristine Bonnevie. We thank the captain and crew of each of these vessels for their hard work as well as the many watch standers who have sailed on the cruises and helped collect the measurements. We also thank Frank Bahr for processing the VMADCP data collected on NRV Alliance and Magnús Danielsen for the processing of the hydrographic data collected on R/V Bjarni Sæmundsson. We acknowledge Leah Trafford McRaven for assistance with Fig. 1 and two anonymous reviewers for their helpful comments, which improved the manuscript. Funding for the project was provided by the Bergen Research Foundation Grant BFS2016REK01 (K. Våge and S. Semper), the Norwegian Research Council under Grant Agreement 231647 (K. Våge), and the U.S. National Science Foundation Grants OCE-1259618 and OCE-1756361 (R. S. Pickart and D. J. Torres), as well as OCE-1558742 (R. S. Pickart). The dataset is available on PANGAEA under https://doi.pangaea.de/10.1594/PANGAEA.903535.
    Keywords: Ocean ; Continental shelf/slope ; Ocean circulation ; Transport ; Intermediate waters ; In situ oceanic observations
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  • 22
    Publication Date: 2022-05-26
    Description: Author Posting. © American Meteorological Society, 2017. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Atmospheric and Oceanic Technology 34 (2017): 1679-1691, doi:10.1175/JTECH-D-16-0162.1.
    Description: For direction-finding high-frequency (HF) radar systems, the correct separation of backscattered spectral energy due to Bragg resonant waves from that due to more complex double-scattering represents a critical first step toward attaining accurate estimates of surface currents from the range-dependent radar backscatter. Existing methods to identify this “first order” region of the spectra, generally sufficient for lower-frequency radars and low-velocity or low-surface gravity wave conditions, are more likely to fail in higher-frequency systems or locations with more variable current, wave, or noise regimes, leading to elevated velocity errors. An alternative methodology is presented that uses a single and globally relevant smoothing length scale, careful pretreatment of the spectra, and marker-controlled watershed segmentation, an image processing technique, to separate areas of spectral energy due to surface currents from areas of spectral energy due to more complex scattering by the wave field or background noise present. Applied to a number of HF radar datasets with a range of operating frequencies and characteristic issues, the new methodology attains a higher percentage of successful first-order identification, particularly during complex current and wave conditions. As operational radar systems continue to expand to more systematically cover areas of high marine traffic, close approaches to ports and harbors, or offshore energy installations, use of this type of updated methodology will become increasingly important to attain accurate current estimates that serve both research and operational interests.
    Description: This analysis was supported by internal funds from the Woods Hole Oceanographic Institution.
    Description: 2018-02-11
    Keywords: Ocean circulation ; Waves, oceanic ; Data processing ; Radars/Radar observations ; Remote sensing ; Pattern detection
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  • 23
    Publication Date: 2022-05-26
    Description: Author Posting. © American Meteorological Society, 2017. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 47 (2017): 2927-2947, doi:10.1175/JPO-D-17-0083.1.
    Description: Motivated by observations in Hudson shelf valley showing stronger onshore than offshore flows, this study investigates wind-driven flows in idealized shallow shelf valleys. This first part of a two-part sequence focuses on the mechanism of the asymmetrical flow response in a valley to along-shelf winds of opposite directions. Model simulations show that (i) when the wind is in the opposite direction to coastal-trapped wave (CTW) phase propagation, the shelf flow turns onshore in the valley and generates strong up-valley transport and a standing meander on the upstream side (in the sense of CTW phase propagation) of the valley, and (ii) when the wind is in the same direction as CTW phase propagation, the flow forms a symmetric onshore detour pattern over the valley with negligible down-valley transport. Comparison of the modeled upstream meanders in the first scenario with CTW characteristics confirms that the up-valley flow results from CTWs being arrested by the wind-driven shelf flow establishing lee waves. The valley bathymetry generates an initial excessive onshore pressure gradient force that drives the up-valley flow and induces CTW lee waves that sustain the up-valley flow. When the wind-driven shelf flow aligns with CTW phase propagation, the initial disturbance generated in the valley propagates away, allowing the valley flow to adjust to roughly follow isobaths. Because of the similarity in the physical setup, this mechanism of arrested CTWs generating stronger onshore than offshore flow is expected to be applicable to the flow response in slope canyons to along-isobath background flows of opposite directions.
    Description: WGZ and SJL were supported by the National Science Foundation through GrantOCE1154575.WGZ is also supported by the NSF Grant OCE 1634965 and SJL by NSF Grant OCE 1558874.
    Description: 2018-06-08
    Keywords: Ocean circulation ; Topographic effects ; Transport ; Vertical motion ; Waves, oceanic ; Wind stress
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  • 24
    Publication Date: 2022-05-26
    Description: Author Posting. © American Meteorological Society, 2014. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Climate 27 (2014): 9359–9376, doi:10.1175/JCLI-D-14-00228.1.
    Description: Multidecadal variability of the Atlantic meridional overturning circulation (AMOC) is examined based on a comparison of the AMOC streamfunctions in depth and in density space, in a 700-yr present-day control integration of the fully coupled Community Climate System Model, version 3. The commonly used depth-coordinate AMOC primarily exhibits the variability associated with the deep equatorward transport that follows the changes in the Labrador Sea deep water formation. On the other hand, the density-based AMOC emphasizes the variability associated with the subpolar gyre circulation in the upper ocean leading to the changes in the Labrador Sea convection. Combining the two representations indicates that the ~20-yr periodicity of the AMOC variability in the first half of the simulation is primarily due to an ocean-only mode resulting from the coupling of the deep equatorward flow and the upper ocean gyre circulation near the Gulf Stream and North Atlantic Current. In addition, the density-based AMOC reveals a gradual change in the deep ocean associated with cooling and increased density, which is likely responsible for the transition of AMOC variability from strong ~20-yr oscillations to a weaker red noise–like multidecadal variability.
    Description: Support from the NOAA Climate Program Office (Grant NA10OAR4310202 and NA13OAR4310139) and NSF EaSM2 (OCE1242989) is gratefully acknowledged.
    Description: 2015-06-15
    Keywords: North Atlantic Ocean ; Meridional overturning circulation ; Ocean circulation ; Thermocline circulation ; Climate variability ; Multidecadal variability
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  • 25
    Publication Date: 2016-05-12
    Description: In this study, we isolated 15 endophytic fungi from five Sudanese medicinal plants. Each fungal endophytic strain was identified by sequencing of internal transcribed spacer (ITS) regions of rDNA. Ethyl acetate extracts were prepared from each endophyte cultivated in vitro and tested for their respective antibacterial activities and antiproliferative activities against human cancer cells. Antibacterial screening was carried out against two bacterial strains: Gram-negative Escherichia coli and Gram-positive methicillin-resistant Staphylococcus aureus , by the broth dilution method. Cell viability was evaluated by the MTT procedure after exposure of MCF7 breast cancer cells and HT29 or HCT116 human colon adenocarcinoma cells to each endophytic extract. Of interest, Byssochlamys spectabilis isolated from Euphorbia prostata showed cytotoxicity (IC 50 = 1.51 ± 0.2 μg mL –1 ) against MCF7 cells, but had a low effect against HT29 or HCT116 cells (IC 50 〉 20 μg mL –1 ). Cladosporium cladosporioides 2, isolated from Vernonia amygdalina leaves, showed antiproliferative activities against MCF7 cells (IC 50 = 10.5 ± 1.5 μg mL –1 ) only. On the other hand, B. spectabilis and Alternaria sp. extract had antibacterial activities against the S. aureus strain. The findings of this work revealed that endophytic fungi associated with medicinal plants from Sudan could be considered as an attractive source of new therapeutic compounds.
    Keywords: Biotechnology & Synthetic Biology
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  • 26
    Publication Date: 2016-04-08
    Description: Resveratrol is a well-known triphenolic natural product present in red wine. For its contribution to human health, the demand for resveratrol as a food and nutrition supplement has increased significantly. In recent years, the rapid development of synthetic biology has promoted extensive work to increase the production of resveratrol in microbes. However, supplementation of expensive phenylpropanoic precursors was required in current engineered strains. Here, we first utilized the site-specific integration strategy to produce resveratrol in Escherichia coli . The genes tal , 4cl and sts were site-specific integrated into the loci of genes tyrR and trpED in the chromosome of E. coli BW25113 (DE3). The final strain was capable of producing 4.612 mg L –1 of resveratrol from glucose.
    Keywords: Biotechnology & Synthetic Biology
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  • 27
    Publication Date: 2016-04-08
    Description: While extensively studied in several model organisms, the role of small, non-coding RNAs in the stress response remains largely unexplored in Clostridium organisms. About 100 years after the first industrial Acetone–Butanol–Ethanol fermentation process, based on the Weizmann Clostridium acetobutylicum strain, strain tolerance to butanol remains a crucial factor limiting the economics of the process. Several studies have examined the response of this organism to metabolite stress, and several genes have been engaged to impart enhanced tolerance, but no sRNAs have yet been directly engaged in this task. We show that the two stress-responsive sRNAs, 6S and tmRNA, upon overexpression impart tolerance to butanol as assessed by viability assays under process-relevant conditions. 6S overexpression enhances cell densities as well as butanol titres. We discuss the likely mechanisms that these two sRNAs might engage in this tolerance phenotype. Our data support the continued exploration of sRNAs as a basis for engineering enhanced tolerance and enhanced solvent production, especially because sRNA-based strategies impose a minimal metabolic burden on the cells.
    Keywords: Biotechnology & Synthetic Biology
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  • 28
    Publication Date: 2016-07-02
    Description: Ten indole alkaloids were obtained from the marine sponge-associated fungus Neosartorya siamensis KUFA 0017. We studied the antimicrobial properties of these and of three other compounds previously isolated from the soil fungus N. siamensis KUFC 6349. Only neofiscalin A showed antimicrobial activity against Gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE); with a minimum inhibitory concentration (MIC) of 8 μg mL –1 against both strains. Another compound, fiscalin C, presented synergistic activity against MRSA when combined with oxacillin, although alone showed no antibacterial effect. Moreover, neofiscalin A, when present at sub-MICs, hampered the ability of both MRSA and VRE strains to form a biofilm. Additionally, the biofilm inhibitory concentration values of neofiscalin A against the MRSA and VRE isolates were 96 and 80 μg mL –1 , respectively. At a concentration of 200 μg mL –1 , neofiscalin A was able to reduce the metabolic activity of the biofilms by ~50%. One important fact is that our results also showed that neofiscalin A had no cytotoxicity against a human brain capillary endothelial cell line.
    Keywords: Biotechnology & Synthetic Biology
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  • 29
    Publication Date: 2016-07-02
    Description: During unconventional protein secretion (UPS), proteins do not pass through the classical endoplasmic reticulum (ER)–Golgi-dependent pathway, but are transported to the cell membrane via alternative routes. One type of UPS is dependent on several autophagy-related (Atg) proteins in yeast and mammalian cells, but mechanisms for unconventional secretion are largely unknown for filamentous fungi. In this study, we investigated whether the autophagy machinery is used for UPS in the filamentous fungus Aspergillus niger . An aspartic protease, which we called PepN, was identified as being likely to be secreted unconventionally, as this protein is highly abundant in culture filtrates during carbon starvation while it lacks a conventional N-terminal secretion sequence. We analysed the presence of PepN in the culture filtrates of carbon starved wild-type, atg1 and atg8 deletion mutant strains by Western blot analysis and by secretome analysis using nanoLC-ESI-MS/MS (wild-type and atg8 deletion mutant). Besides the presence of carbohydrate-active enzymes and other types of proteases, PepN was abundantly found in culture filtrates of both wild-type and atg deletion strains, indicating that the secretion of PepN is independent of the autophagy machinery in A. niger and hence most likely occurs via a different mechanism.
    Keywords: Biotechnology & Synthetic Biology
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  • 30
    Publication Date: 2016-08-27
    Description: Actin-like MreB paralogs play important roles in cell shape maintenance, cell wall synthesis and the regulation of the D,L-endopeptidases, CwlO and LytE. The gram-positive bacteria, Bacillus amyloliquefaciens LL3, is a poly--glutamic acid (-PGA) producing strain that contains three MreB paralogs: MreB, Mbl and MreBH. In B. amyloliquefaciens , CwlO and LytE can degrade -PGA. In this study, we aimed to test the hypothesis that modulating transcript levels of MreB paralogs would alter the synthesis and degradation of -PGA. The results showed that overexpression or inhibition of MreB, Mbl or MreBH had distinct effects on cell morphology and the molecular weight of the -PGA products. In fermentation medium, cells of mreB inhibition mutant were 50.2% longer than LL3, and the -PGA titer increased by 55.7%. However, changing the expression level of mbl showed only slight effects on the morphology, -PGA molecular weight and titer. In the mreBH inhibition mutant, -PGA production and its molecular weight increased by 56.7% and 19.4%, respectively. These results confirmed our hypothesis that suppressing the expression of MreB paralogs might reduce -PGA degradation, and that improving the cell size could strengthen -PGA synthesis. This is the first report of enhanced -PGA production via suppression of actin-like MreB paralogs.
    Keywords: Biotechnology & Synthetic Biology
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  • 31
    Publication Date: 2016-06-08
    Description: Microbial production of acetone and butanol was one of the first large-scale industrial fermentation processes of global importance. During the first part of the 20th century, it was indeed the second largest fermentation process, superseded in importance only by the ethanol fermentation. After a rapid decline after the 1950s, acetone-butanol-ethanol (ABE) fermentation has recently gained renewed interest in the context of biorefinery approaches for the production of fuels and chemicals from renewable resources. The availability of new methods and knowledge opens many new doors for industrial microbiology, and a comprehensive view on this process is worthwhile due to the new interest. This thematic issue of FEMS Microbiology Letters, dedicated to the 100th anniversary of the first industrial exploitation of Chaim Weizmann's ABE fermentation process, covers the main aspects of old and new developments, thereby outlining a model development in biotechnology. All major aspects of industrial microbiology are exemplified by this single process. This includes new technologies, such as the latest developments in metabolic engineering, the exploitation of biodiversity and discoveries of new regulatory systems such as for microbial stress tolerance, as well as technological aspects, such as bio- and down-stream processing.
    Keywords: Biotechnology & Synthetic Biology
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  • 32
    Publication Date: 2016-06-02
    Description: Food-grade lactic acid bacteria, such as lactobacilli, represent good candidates for the development of mucosal vectors. Indeed, they are generally recognized as safe microorganisms and some strains display beneficial effects (probiotics). In this study, we described a new lactobacilli in vivo expression (LIVE) system for the production and delivery of therapeutic molecules at mucosal surfaces. The versatility and functionality of this system was successfully validated in several lactobacilli species; furthermore, we assessed in vivo LIVE system in two different mouse models of human pathologies: (i) a model of therapy against intestinal inflammation (inflammatory bowel diseases) and (ii) a model of vaccination against dental caries. We demonstrated that Lactobacillus gasseri expressing the anti-inflammatory cytokine IL-10 under LIVE system efficiently delivered the recombinant protein at mucosal surfaces and display anti-inflammatory effects. In the vaccination model against caries, LIVE system allowed the heterologous expression of Streptococcus mutans antigen GbpB by L. gasseri , leading to a stimulation of the host immune response.
    Keywords: Biotechnology & Synthetic Biology
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  • 33
    Publication Date: 2016-06-02
    Description: The microbial fermentation process is one of the sustainable and environment-friendly ways to produce 1-butanol and other bio-based chemicals. The success of the fermentation process greatly relies on the choice of bioreactors and the separation methods. In this review, the history and the performance of bioreactors for the acetone–butanol–ethanol (ABE) fermentation is discussed. The subject is then focused on in situ product recovery (ISPR) techniques, particularly for the integrated extraction-gas stripping. The usefulness of this promising hybrid ISPR device is acknowledged by its incorporation with batch, fed-batch and continuous processes to improve the performance of ABE fermentation.
    Keywords: Biotechnology & Synthetic Biology
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  • 34
    Publication Date: 2016-01-29
    Description: Anaerobic Clostridium spp. is an important bioproduction microbial genus that can produce solvents and utilize a broad spectrum of substrates including cellulose and syngas. Genome-scale metabolic (GSM) models are increasingly being put forth for various clostridial strains to explore their respective metabolic capabilities and suitability for various bioconversions. In this study, we have selected representative GSM models for six different clostridia ( Clostridium acetobutylicum , C. beijerinckii , C. butyricum , C. cellulolyticum , C. ljungdahlii and C. thermocellum ) and performed a detailed model comparison contrasting their metabolic repertoire. We also discuss various applications of these GSM models to guide metabolic engineering interventions as well as assessing cellular physiology.
    Keywords: Biotechnology & Synthetic Biology
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  • 35
    Publication Date: 2016-03-02
    Description: Biobutanol is a next-generation liquid biofuel with properties akin to those of gasoline. There is a widespread effort to commercialize biobutanol production from agricultural residues, such as corn stover, which do not compete with human and animal foods. This pursuit is backed by extensive government mandates to expand alternative energy sources. This review provides an overview of research on biobutanol production using corn stover feedstock. Structural composition, pretreatment, sugar yield (following pretreatment and hydrolysis) and generation of lignocellulose-derived microbial inhibitory compounds (LDMICs) from corn stover are discussed. The review also discusses different Clostridium species and strains employed for biobutanol production from corn stover-derived sugars with respect to solvent yields, tolerance to LDMICs and in situ solvent recovery (integrated fermentation). Further, the economics of cellulosic biobutanol production are highlighted and compared to corn starch-derived ethanol and gasoline. As discussed herein, the economic competitiveness of biobutanol production from corn stover largely depends on feedstock processing and fermentation process design.
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  • 36
    Publication Date: 2016-02-20
    Description: Biological production in heterologous hosts is of interest for the production of the C4 alcohol (butanol) and other chemicals. However, some hurdles need to be overcome in order to achieve an economically viable process; these include avoiding the consumption of butanol and maintaining tolerance to this solvent during production. Pseudomonas putida is a potential host for solvent production; in order to further adapt P. putida to this role, we generated mini-Tn 5 mutant libraries in strain BIRD-1 that do not consume butanol. We analyzed the insertion site of the mini-Tn 5 in a mutant that was deficient in assimilation of butanol using arbitrary PCR followed by Sanger sequencing and found that the transposon was inserted in the malate synthase B gene. Here, we show that in a second round of mutagenesis a double mutant unable to take up butanol had an insertion in a gene coding for a multisensor hybrid histidine kinase. The genetic context of the histidine kinase sensor revealed the presence of a set of genes potentially involved in butanol assimilation; qRT-PCR analysis showed induction of this set of genes in the wild type and the malate synthase mutant but not in the double mutant.
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  • 37
    Publication Date: 2016-02-20
    Description: The heavy dependence on petroleum-derived fuel has raised concerns about energy sustainability and climate change, which have prompted researchers to explore fuel production from renewable sources. 1-Butanol and isobutanol are promising biofuels that have favorable properties and can also serve as solvents or chemical feedstocks. Microbial production of these alcohols provides great opportunities to access a wide spectrum of renewable resources. In recent years, research has improved the native 1-butanol production and has engineered isobutanol production in various organisms to explore metabolic diversity and a broad range of substrates. This review focuses on progress in metabolic engineering for the production of these two compounds using various resources.
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  • 38
    Publication Date: 2016-02-20
    Description: Biobased production of butanol promises a more sustainable route for industrial production. However, butanol toxicity remains a barrier for achieving high product titers. Investigation into butanol stress has shed some light on its modes of toxicity. Unfortunately, there still remain significant shortfalls in our understanding of the complex interactions of butanol with cells. To address this knowledge gap, a diverse range of tools have been employed to gain a better understanding of the adverse effects of butanol on the cell. These findings have lead to the identification of possible molecular mechanisms associated with butanol tolerance, which can be harnessed for future strain development efforts. This review focuses on recent efforts to address the toxicity of butanol in microbial producers and offers some perspectives on the future direction of this research sector.
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  • 39
    Publication Date: 2016-02-25
    Description: Biobutanol outperforms bioethanol as an advanced biofuel, but is not economically competitive in terms of its titer, yield and productivity associated with feedstocks and energy cost. In this work, the synergistic effect of calcium and zinc was investigated in the acetone-butanol-ethanol (ABE) fermentation by Clostridium acetobutylicum using glucose, xylose and glucose/xylose mixtures as carbon source(s). Significant improvements associated with enhanced glucose/xylose utilization, cell growth, acids re-assimilation and butanol biosynthesis were achieved. Especially, the maximum butanol and ABE production of 16.1 and 25.9 g L –1 were achieved from 69.3 g L –1 glucose with butanol/ABE productivities of 0.40 and 0.65 g L –1 h –1 compared to those of 11.7 and 19.4 g/L with 0.18 and 0.30 g L –1 h –1 obtained in the control respectively without any supplement. More importantly, zinc was significantly involved in the butanol tolerance based on the improved xylose utilization under various butanol-shock conditions (2, 4, 6, 8 and 10 g L –1 butanol). Under the same conditions, calcium and zinc co-supplementation led to the best xylose utilization and butanol production. These results suggested that calcium and zinc could play synergistic roles improving ABE fermentation by C. acetobutylicum .
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  • 40
    Publication Date: 2016-02-07
    Description: Pseudomonas putida KT2440 is a saprophytic, environmental microorganism that plays important roles in the biodegradation of environmental toxic compounds and production of polymers, chemicals and secondary metabolites. Gene deletion of KT2440 usually involves cloning of the flanking homologous fragments of the gene of interest into a suicide vector followed by transferring into KT2440 via triparental conjugation. Selection and counterselection steps are then employed to generate gene deletion mutant. However, these methods are tedious and are not suitable for the manipulation of multiple genes simultaneously. Herein, a two-step, markerless gene deletion method is presented. First, homologous armsflanked loxP-neo-loxP was knocked-in to replace the gene of interest, then the kanamycin resistance marker is removed by Cre recombinase catalyzed site-specific recombination. Both two-plasmid and one-plasmid gene systems were established. MekR/P mekA regulated gene expression system was found to be suitable for tight Cre expression in one-plasmid deletion system. The straightforward, time saving and highly efficient markerless gene deletion strategy has the potential to facilitate the genetics and functional genomics study of P. putida KT2440.
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  • 41
    Publication Date: 2016-03-02
    Description: Flow cytometry, in combination with fluorescent staining, was used to evaluate population heterogeneity in acetone-butanol–ethanol fermentation that was carried out with type strain Clostridium beijerinckii NCIMB 8052 and non-type C. pasteurianum NRRL B-598. A combination of propidium iodide (PI) and carboxyfluorescein diacetate (CFDA), PI plus Syto-9 and bis-oxonol (BOX) alone were employed to distinguish between active and damaged cells together with simultaneous detection of spores. These strategies provided valuable information on the physiological state of clostridia. CFDA and PI staining gave the best separation of four distinct subpopulations of enzymatically active cells, doubly stained cells, damaged cells and spores. Proportional representation of cells in particular sub-regions correlated with growth characteristics, fermentation parameters such as substrate consumption and product formation in both species under different cultivation conditions.
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  • 42
    Publication Date: 2016-02-03
    Description: Simultaneous saccharification and fermentation (SSF) process was applied for biobutanol production by Clostridium saccharobutylicum DSM 13864 from corn stover (CS). The key influential factors in SSF process, including corn steep liquor concentration, dry biomass and enzyme loading, SSF temperature, inoculation size and pre-hydrolysis time were optimized. In 5-L bioreactor with SSF process, butanol titer and productivity of 12.3 g/L and 0.257 g/L/h were achieved at 48 h, which were 20.6% and 21.2% higher than those in separate hydrolysis and fermentation (SHF), respectively. The butanol yield reached 0.175 g/g pretreated CS in SSF, representing 50.9% increase than that in SHF (0.116 g/g pretreated CS). This study proves the feasibility of efficient and economic production of biobutanol from CS by SSF.
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  • 43
    Publication Date: 2016-03-13
    Description: We previously reported that the β-1,4-Mannanase ( manB ) gene from Bacillus pumilus functions as a good reporter gene in Lactobacillus casei . Two vectors were constructed. One carries the signal peptide of secretion protein Usp45 (SP Usp45 ) from Lactococcus lactis (pELSH), and the other carries the full-length S-layer protein, SlpA, from L. acidophilus (pELWH). In this work, another vector, pELSPH, was constructed to include the signal peptide of protein SlpA (SP SlpA ), and the capacity of all three vectors to drive expression of the manB gene in L. casei was evaluated. The results showed that SP Usp45 is functionally recognized and processed by the L. casei secretion machinery. The SP Usp45 -mediated secretion efficiency was ~87%, and SP SlpA drove the export of secreted ManB with ~80% efficiency. SP SlpA secretion was highly efficient, and expressed SlpA was anchored to the cell wall by an unknown secretion mechanism. Full-length SlpA drove the cell wall-anchored expression of an SlpA-ManB fusion protein but at a much lower level than that of protein SlpA.
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  • 44
    Publication Date: 2016-03-13
    Description: The porin MspA of Mycobacterium smegmatis is a biological nanopore used for DNA sequencing. The octameric MspA pore can be isolated from M. smegmatis in milligram quantities, is extremely stable against denaturation and rapidly inserts into lipid membranes. Here, we show that MspA pores composed of different Msp subunits are formed in M. smegmatis and that hetero-oligomers of different Msp monomers increase the heterogeneity of MspA pores designed for DNA sequencing. To improve the quality of preparations of mutant MspA proteins, all four msp genes were deleted from the M. smegmatis genome after insertion of an inducible porin gene from M. tuberculosis. In the msp quadruple mutant M. smegmatis ML712 no Msp porins were detected and mutant MspA proteins were produced at wild-type levels. Lipid bilayer experiments demonstrated that MspA pores isolated from ML712 formed functional channels and had a narrower conductance distribution than pores purified from M. smegmatis with background msp expression. Thus, the M. smegmatis msp quadruple mutant improves the homogeneity of MspA pores designed for DNA sequencing and might also facilitate the identification and functional characterization of other mycobacterial pore proteins.
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  • 45
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2016-03-04
    Description: Mostly, butanol is formed as a product by saccharolytic anaerobes, employing the so-called ABE fermentation (for acetone–butanol–ethanol). However, this alcohol can also be produced from gaseous substrates such as syn(thesis) gas (major components are carbon monoxide and hydrogen) by autotrophic acetogens. In view of economic considerations, a biotechnological process based on cheap and abundant gases such as CO and CO 2 as a carbon source is preferable to more expensive sugar or starch fermentation. In addition, any conflict for use of substrates that can also serve as human nutrition is avoided. Natural formation of butanol has been found with, e.g. Clostridium carboxidivorans , while metabolic engineering for butanol production was successful using, e.g. C. ljungdahlii . Production of butanol from CO 2 under photoautotrophic conditions was also possible by recombinant DNA construction of a respective cyanobacterial Synechococcus sp. PCC 7942 strain.
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  • 46
    Publication Date: 2016-05-20
    Description: Lichen-forming fungi and extracts derived from them have been used as alternative medicine sources for millennia and recently there has been a renewed interest in their known bioactive properties for anticancer agents, cosmetics and antibiotics. Although lichen-forming fungus-derived compounds are biologically and commercially valuable, few studies have been performed to determine their modes of action. This study used chemical-genetic and chemogenomic high-throughput analyses to gain insight into the modes of action of Caloplaca flavoruscens extracts. High-throughput screening of 575 lichen extracts was performed and 39 extracts were identified which inhibited yeast growth. A C. flavoruscens extract was selected as a promising antifungal and was subjected to genome-wide haploinsufficiency profiling and homozygous profiling assays. These screens revealed that yeast deletion strains lacking Rsc8, Pro1 and Toa2 were sensitive to three concentrations (IC 25.5 , IC 25 and IC 50 , respectively) of C. flavoruscens extract. Gene-enrichment analysis of the data showed that C. flavoruscens extracts appear to perturb transcription and chromatin remodeling.
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  • 47
    Publication Date: 2016-05-20
    Description: Plant lectins, which are proteins/glycoproteins present in a wide range of vegetables, fruits, cereals and beans, are resistant to digestive enzymes and food cooking temperatures. They bind reversibly to specific glycosidic residues expressed on the membrane of intestinal epithelial cells (IEC) and cause anti-nutritional effects in humans and animals. Soybean lectin (SBA) has been detected in poultry diets, and its ability to bind to the intestinal epithelium has been reported. The development of new methods for removing SBA from feeds or to prevent interaction with the intestinal mucosa is of interest. In this study, the in vitro cytotoxicity of SBA on IEC of chicks was demonstrated for the first time. The LD 50 , assessed after 2 h exposure of IEC to SBA, was 6.13 μg mL –1 . The ability of Bifidobacterium infantis CRL1395 to bind SBA on the bacterial envelope was confirmed, and prevention of IEC cytotoxicity by lectin removal was demonstrated. Safety of B. infantis CRL1395, resistance to gastrointestinal stress and adhesion were also determined. It was concluded that the early administration of B. infantis CRL1395 to chicks would effectively reduce the toxicity of SBA. Besides, it would favour the colonization of the gut with a beneficial microbiota.
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  • 48
    Publication Date: 2016-04-08
    Description: Fermentation-based production of butyric acid is robust and efficient. Modern catalytic technologies make it possible to convert butyric acid to important fine chemicals and biofuels. Here, current chemocatalytic and biocatalytic conversion methods are reviewed with a focus on upgrading butyric acid to 1-butanol or butyl-butyrate. Supported Ruthenium- and Platinum-based catalyst and lipase exhibit important activities which can pave the way for more sustainable process concepts for the production of green fuels and chemicals.
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  • 49
    Publication Date: 2016-04-15
    Description: Clostridium acetobutylicum is an industrially important Gram-positive organism, which is capable of producing economically important chemicals in the ABE (Acetone, Butanol and Ethanol) fermentation process. Renewed interests in the ABE process necessitate the availability of additional genetics tools to facilitate the derivation of a greater understanding of the underlying metabolic and regulatory control processes in operation through forward genetic strategies. In this study, a xylose inducible, mariner -based, transposon system was developed and shown to allow high-efficient random mutagenesis in the model strain ATCC 824. Of the thiamphenicol resistant colonies obtained, 91.9% were shown to be due to successful transposition of the catP- based mini-transposon element. Phenotypic screening of 200 transposon clones revealed a sporulation-defective clone with an insertion in spo0A , thereby demonstrating that this inducible transposon system can be used for forward genetic studies in C. acetobutylicum .
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  • 50
    Publication Date: 2016-04-15
    Description: Bacteria are becoming increasingly resistant to currently used antibiotics. At the same time, little progress has been made in discovering new antibacterial drugs to combat resistant organisms. History teaches us that ‘high tech’ target-based complex methods are not synonymous with success and a return to simple, systematic screening of natural products against bacteria from traditional and novel resources holds our greatest hope of success.
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  • 51
    Publication Date: 2016-04-15
    Description: The economic upturn of the past 200 years would not have been conceivable without fossil resources such as coal and oil. However, the fossil-based economy increasingly reaches its limits and displays contradictions. Bioeconomy, strategically combining economy and ecology willing to make biobased and sustainable growth possible, is promising to make a significant contribution towards solving these issues. In this context, microbial bioconversions are promising to support partially the increasing need for materials and fuels starting from fresh, preferably waste, biomass. Butanol is a very attractive molecule finding applications both as a chemical platform and as a fuel. Today it principally derives from petroleum, but it also represents the final product of microbial catabolic pathways. Because of the need to maximize yield, titer and productivity to make the production competitive and viable, the challenge is to transform a robustly regulated metabolic network into the principal cellular activity. However, this goal can only be accomplished by a profound understanding of the cellular physiology, survival strategy and sensing/signalling cascades. Here, we shortly review on the natural cellular pathways and circumstances that lead to n -butanol accumulation, its physiological consequences that might not match industrial needs and on possible solutions for circumventing these natural constraints.
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  • 52
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2016-04-20
    Description: Butanol is a very interesting substance both for the chemical industry and as a biofuel. The classical distillation process for the removal of butanol is far too energy demanding, at a factor of 220% of the energy content of butanol. Alternative separation processes studied are hybrid processes of gas-stripping, liquid–liquid extraction and pervaporation with distillation and a novel adsorption/drying/desorption hybrid process. Compared with the energy content of butanol, the resulting energy demand for butanol separation and concentration of optimized hybrid processes is 11%–22% for pervaporation/distillation and 11%–17% for liquid–liquid extraction/distillation. For a novel adsorption/drying/desorption process, the energy demand is 9.4%. But all downstream process options need further proof of industrial applicability.
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  • 53
    Publication Date: 2016-04-20
    Description: Acetone–butanol–ethanol (ABE) fermentation is a metabolic process of clostridia that produces bio-based solvents including butanol. It is enabled by an underlying metabolic reaction network and modulated by cellular gene regulation and environmental cues. Mathematical modeling has served as a valuable strategy to facilitate the understanding, characterization and optimization of this process. In this review, we highlight recent advances in system-level, quantitative modeling of ABE fermentation. We begin with an overview of integrative processes underlying the fermentation. Next we survey modeling efforts including early simple models, models with a systematic metabolic description, and those incorporating metabolism through simple gene regulation. Particular focus is given to a recent system-level model that integrates the metabolic reactions, gene regulation and environmental cues. We conclude by discussing the remaining challenges and future directions towards predictive understanding of ABE fermentation.
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  • 54
    Publication Date: 2016-02-10
    Description: Cry proteins are pesticidal toxins produced by the bacterium Bacillus thuringiensis (Bt), which aggregate in sporulating cells to form a crystal. Except in a relatively few cases, these crystals are located outside the exosporium that surrounds the spore. Bt2-56 is a strain of Bt that has the relatively uncommon characteristic of locating its Cry protein-containing crystal within the exosporium, and in association with a long, multifiber filament. With the ultimate goal of both understanding and manipulating the localization of Cry proteins within the exosporium, we sought to identify the genes coding for the exosporium-localized Cry proteins in Bt2-56. Herein we show (i) that five cry-like genes are present in the genome of Bt2-56, (ii) that two pairs of these genes show organizational similarity to a relatively uncommon gene configuration that coexpress a cry gene along with a gene whose product aids crystal formation and (iii) that when one of these two gene pairs (cry21A-cdA) is expressed in an acrystalliferous strain of Bt, crystals are formed that localize within the exosporium. In Bt ssp. finitimus , the only other strain in which crystal localization has been studied, a Cry protein needed expression of two non-cry ORFs in order to localize within the exosporium, indicating that there are some mechanistic differences for crystal localization between Bt ssp. finitimus and Bt2-56.
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  • 55
    Publication Date: 2016-02-12
    Description: The commercialization of the n -butanol bioprocess is largely dependent on the price of feedstocks. Renewable cellulose appears to be an appealing feedstock. The microbial production of n -butanol still remains challenging because of the limited availability of intracellular NADH. To address this issue, an Escherichia coli strain carrying the clostridial CoA-dependent pathway was supplied with heterologous formate dehydrogenase. With the cellulose hydrolysate of rice straw, this single strain produced cellulosic biobutanol with a production yield at 35% of the theoretical and a productivity of 0.093 g L –1  h –1 . In an alternative method, the production involved a co-culture system consisting of two separate strains provided with the full CoA-dependent pathway. This system achieved a production yield and productivity reaching 62.8% of the theoretical and 0.163 g L –1  h –1 , respectively. The result indicates that the E. coli co-culture system is technically promising for the production of cellulosic biobutanol.
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  • 56
    Publication Date: 2016-11-09
    Description: Hepatitis E virus (HEV) is the pathogen causing hepatitis E (HE). It arouses global public health concern since it is a zoonotic disease. The objective of this letter is to report a cost-effective internal control prepared for monitoring procedures of HEV reverse transcriptase (RT)-PCR detection. A selected conserved HEV RNA fragment was integrated into the downstream of the truncated MS2 bacteriophage genome based on Armored RNA technology. The resulting MS2-HEV gene harbored by the pET-28b-MS2-HEV plasmid was transformed into E. coli BL21(DE3) for expression analysis by SDS-PAGE. The expression products were purified and concentrated by ultrasonication and ultrafiltration separation. The morphology and stability properties of the virus-like particles (VLPs) were evaluated by electron microscopy scanning and nuclease challenges, respectively. SDS-PAGE results showed that the constructed MS2-HEV gene expressed efficiently and the purity of the VLPs was highly consistent with the result in electron microscopy. Stability evaluation results demonstrated that the prepared VLPs exhibited strong resistance to DNase I and RNase A attacks and also performed long-lasting protection of coated HEV RNA for at least 4 months at –20°C. These data revealed that the prepared VLPs meet the basic requirements of use as internal control material in the HEV RNA amplification assay.
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  • 57
    Publication Date: 2016-09-08
    Description: Epigenetics is the study of heritable changes in gene expression without concomitant changes in DNA sequence. Due to its relevance in development, differentiation and human health, epigenetics has recently become an emerging area of science with regard to eukaryotic organisms and has shown enormous potential in synthetic biology. However, significant examples of epigenetic regulation in bacterial synthetic biology have not yet been reported. In the current study, we present the first model of such an epigenetic circuit. We termed the circuit the alternator circuit because parental cells carrying this circuit and their progeny alternate between distinct and heritable cellular fates without undergoing changes in genome sequence. Furthermore, we demonstrated that the alternator circuit exhibits hysteresis because its output depends not only on its present state but also on its previous states.
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  • 58
    Publication Date: 2016-10-12
    Description: Bacteriophages produce endolysins (peptidoglycan hydrolases) to lyse the host cell from within and release nascent bacteriophage particles. Recombinant endolysins can lyse Gram-positive bacteria when added exogenously. As a potential alternative antimicrobial, we cloned and expressed the enterococcal VD13 bacteriophage endolysin. VD13 endolysin has a CHAP catalytic domain with 92% identity with the bacteriophage IME-EF1 endolysin. The predicted size of VD13 endolysin is ~27 kDa as verified by SDS-PAGE. The VD13 endolysin lyses Enterococcus faecalis strains, but not E. faecium or other non-enterococci. VD13 endolysin has activity from pH 4 to pH 8, with peak activity at pH 5, and exhibits greater activity in the presence of calcium. Optimum activity at pH 5 occurs in the absence of NaCl. VD13 endolysin, in ammonium acetate (C 2 H 3 O 2 NH 4 ) calcium chloride (CaCl 2 ) buffer pH 5, is stimulated to higher activity upon heating at temperatures up to 65°C for 30 min, whereas activity is lost upon heating to 42°C, in pH 7 buffer.
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  • 59
    Publication Date: 2016-09-08
    Description: Cultivation in a bioreactor of immobilized deep-sea hydrothermal microbial community was tested in order to assess the stability and reactivity of this new system. A community composed of eight hydrothermal strains was entrapped in a polymer matrix that was used to inoculate a continuous culture in a gas-lift bioreactor. The continuous culture was performed for 41 days at successively 60°C, 55°C, 60°C, 85°C and 60°C, at pH 6.5, in anaerobic condition and constant dilution rate. Oxic stress and pH variations were tested at the beginning of the incubation. Despite these detrimental conditions, three strains including two strict anaerobes were maintained in the bioreactor. High cell concentrations (3 x 10 8 cells mL –1 ) and high ATP contents were measured in both liquid fractions and beads. Cloning-sequencing and qPCR revealed that Bacillus sp. dominated at the early stage, and was later replaced by Thermotoga maritima and Thermococcus sp. Acetate, formate and propionate concentrations varied simultaneously in the liquid fractions. These results demonstrate that these immobilized cells were reactive to culture conditions. They were protected inside the beads during the stress period and released in the liquid fraction when conditions were more favorable. This confirms the advantage of immobilization that highlights the resilience capacity of certain hydrothermal microorganisms after a stress period.
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  • 60
    Publication Date: 2016-10-12
    Description: To study the viability of a gyrA S83 stop mutation found in an Escherichia coli J53 ciprofloxacin-resistant strain (J53 CipR27), a pBR322 derivative was constructed with a TAG mutation in the bla gene knocking out ampicillin resistance. Ampicillin resistance was restored, suggesting that the strain contains tRNA suppressor activity able to suppress the UAG codon gyrA and allow viability. The method was applied to 22 unique clinical E. coli isolates, and all were found to have low-level suppressor activity.
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  • 61
    Publication Date: 2016-11-17
    Description: Enterotoxigenic Escherichia coli (ETEC) bacteria producing heat-stable toxin (STa) and/or heat-labile toxin (LT) are among top causes of children's diarrhea and travelers’ diarrhea. Currently no vaccines are available for ETEC associated diarrhea. A major challenge in developing ETEC vaccines is the inability to stimulate protective antibodies against the key STa toxin that is potently toxic and also poorly immunogenic. A recent study suggested toxoid fusion 3xSTa N12S -dmLT, which consists of a monomer LT toxoid (LT R192G/L211A ) and three copies of STa toxoid STa N12S , may represent an optimal immunogen inducing neutralizing antibodies against STa toxin [IAI 2014, 82(5):1823-32]. In this study, we immunized mice with this fusion protein following a different parenteral route and using different adjuvants to further characterize immunogenicity of this toxoid fusion. Data from this study showed that 3xSTa N12S -dmLT toxoid fusion induced neutralizing anti-STa antibodies in the mice following subcutaneous immunization, as effectively as in the mice under intraperitoneal route. Data also indicated that double mutant LT (dmLT) can be an effective adjuvant for this toxoid fusion in mice subcutaneous immunization. Results from this study affirmed that toxoid fusion 3xSTa N12S -dmLT induces neutralizing antibodies against STa toxin, suggesting this toxoid fusion is potentially a promising immunogen for ETEC vaccine development.
    Keywords: Biotechnology & Synthetic Biology
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    Electronic ISSN: 1574-6968
    Topics: Biology
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  • 62
    Publication Date: 2016-12-23
    Description: Carocin D is a bacteriocin produced by Pectobacterium carotovorum subsp. carotovorum Pcc21. Carocin D inhibits the growth of P . carotovorum subsp. carotovorum and closely related strains. Pectobacterium carotovorum subsp. carotovorum is a causative bacterium for soft rot disease and leads to severe economic losses. Bacteriocins recognize and interact with a specific membrane protein of target bacteria as a receptor. To identify the receptor responsible for carocin D recognition, mutants that underwent a phenotypic change from carocin D sensitivity to carocin D insensitivity were screened. Based on Tn 5 insertions, carocin D sensitivity was dependent on expression of the outer membrane protein OmpF. The insensitivity of the mutant (Pcc3MR) to carocin D was complemented with ompF from carocin D-sensitive strains, not from carocin D-resistant strains. The selectivity between sensitive and resistant strains could be attributed to variation in OmpFs in the cell-surface-exposed regions. Based on sequence analysis and complementation assays, it appears that carocin D uses OmpF as a receptor and is translocated by the TonB system. According to previously reported translocation mechanisms of colicins, OmpF works along with the TolA system rather than the TonB system. Therefore, the current findings suggest that carocin D is imported by a unique colicin-like bacteriocin translocation system.
    Keywords: Biotechnology & Synthetic Biology
    Print ISSN: 0378-1097
    Electronic ISSN: 1574-6968
    Topics: Biology
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  • 63
    Publication Date: 2016-11-17
    Description: Pseudomonas putida KT2440 is a saprophytic and generally recognized as safe microorganism that plays important roles in the biodegradation and production of value-added chemicals. Chromosomal gene deletion of P. putida KT2440 usually involves time-consuming gene cloning, conjugal transfer and counterselection. Recently, we developed a P. putida KT2440 markerless gene deletion method based on recombineering and Cre/ loxP site-specific recombination. PCR-based Red recombineering circumvents the tedious cloning steps and is more amenable to high-throughput manipulation. Here we report an improved scarless gene deletion strategy based on recombineering and intron-encoded homing endonuclease I-SceI-mediated double-strand break repair. Sixteen drug exporter gene(s) were deleted and the minimal inhibition concentrations of the mutants to a variety of antibiotics were determined. The robustness of the procedure was also demonstrated by sequential deletion of five large genomic regions. Up to 59% recombination efficiency was achieved for a 54.8 kb deletion, and the efficiency of RecA-mediated double-strand break repair, which was boosted by Red recombinase, was nearly 100%. The strain with a 3.76% genome reduction showed an improved growth rate and transformation efficiency. The straightforward, time-saving and highly efficient scarless deletion approach has the potential to facilitate the genetic study, and biotechnological and environmental applications of P. putida KT2440.
    Keywords: Biotechnology & Synthetic Biology
    Print ISSN: 0378-1097
    Electronic ISSN: 1574-6968
    Topics: Biology
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  • 64
    Publication Date: 2016-12-16
    Description: Human health has been seriously endangered by highly prevalent salmonellosis and multidrug-resistant Salmonella strains. Current vaccines suffer from variable immune-protective effects, so more effective ones are needed to control Salmonella infection . Bacterial ghosts have been produced by the expression of lysis gene E from bacteriophage PhiX174 and can be filled with considerable exogenous substances such as DNA or drugs as a novel platform. In this study, Salmonella enteritidis (SE) ghosts were developed and loaded with Neisseria gonorrhoeae porin B (porB) to construct a novel inactive vaccine. Our new studies show that SE ghosts loaded with porB displayed increased production of pro-inflammatory cytokines (IL-1β, IL-6, IL-10 and IL-12p70) in bone marrow-derived dendritic cells (BMDCs), and elicited significantly higher specific systemic and mucosal immune responses to Salmonella than SE ghosts alone. In addition, the novel porB-loaded ghosts conferred higher protective effects on virulent Salmonella challenge. For the first time, we demonstrate that N. gonorrhoeae porB, as a novel adjuvant, can increase the immunogenicity of SE ghosts. Our studies suggested that Salmonella enteritidis ghosts loaded with Neisseria gonorrhoeae porin B might be a useful mucosal Salmonella vaccine candidate for practical use in the future.
    Keywords: Biotechnology & Synthetic Biology
    Print ISSN: 0378-1097
    Electronic ISSN: 1574-6968
    Topics: Biology
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  • 65
    Publication Date: 2016-12-12
    Description: The food-grade bacterium Lactococcus lactis is increasingly used for heterologous protein expression in therapeutic and industrial applications. The ability of L. lactis to secrete biologically active cytokines may be used for the generation of therapeutic cytokines. Interleukin (IL)-18 enhances the immune response, especially on mucosal surfaces, emphasizing its therapeutic potential. However, it is produced as an inactive precursor and has to be enzymatically cleaved for maturation. We genetically manipulated L. lactis to secrete murine IL-18. The mature murine IL-18 gene was inserted downstream of a nisin promoter in pNZ8149 plasmid and the construct was used to transform L. lactis NZ3900 . The transformants were selected on Elliker agar and confirmed by restriction enzyme digestion and sequencing. The expression and secretion of IL-18 protein was verified by SDS-PAGE, western blotting and ELISA. The biological activity of recombinant IL-18 was determined by its ability to induce interferon (IFN)- production in L. lactis co-cultured with murine splenic T cells. The amounts of IL-18 in bacterial lysates and supernatants were 3–4 μg mL –1 and 0.6–0.7 ng mL –1 , respectively. The successfully generated L. lactis strain that expressed biologically active murine IL-18 can be used to evaluate the possible therapeutic effects of IL-18 on mucosal surfaces.
    Keywords: Biotechnology & Synthetic Biology
    Print ISSN: 0378-1097
    Electronic ISSN: 1574-6968
    Topics: Biology
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