scholarly journals Cross-utilization of β-galactosides and cellobiose in Geobacillus stearothermophilus

2020 ◽  
Vol 295 (31) ◽  
pp. 10766-10780
Author(s):  
Smadar Shulami ◽  
Arie Zehavi ◽  
Valery Belakhov ◽  
Rachel Salama ◽  
Shifra Lansky ◽  
...  
Keyword(s):  
Transport System ◽  
Linear Growth ◽  
Growth Curves ◽  
Thermophilic Bacterium ◽  
Transport Systems ◽  
Geobacillus Stearothermophilus ◽  
Phosphotransferase System ◽  
Sensing System ◽  
Galactose Transport ◽  
Logarithmic Growth

Strains of the Gram-positive, thermophilic bacterium Geobacillus stearothermophilus possess elaborate systems for the utilization of hemicellulolytic polysaccharides, including xylan, arabinan, and galactan. These systems have been studied extensively in strains T-1 and T-6, representing microbial models for the utilization of soil polysaccharides, and many of their components have been characterized both biochemically and structurally. Here, we characterized routes by which G. stearothermophilus utilizes mono- and disaccharides such as galactose, cellobiose, lactose, and galactosyl-glycerol. The G. stearothermophilus genome encodes a phosphoenolpyruvate carbohydrate phosphotransferase system (PTS) for cellobiose. We found that the cellobiose-PTS system is induced by cellobiose and characterized the corresponding GH1 6-phospho-β-glucosidase, Cel1A. The bacterium also possesses two transport systems for galactose, a galactose-PTS system and an ABC galactose transporter. The ABC galactose transport system is regulated by a three-component sensing system. We observed that both systems, the sensor and the transporter, utilize galactose-binding proteins that also bind glucose with the same affinity. We hypothesize that this allows the cell to control the flux of galactose into the cell in the presence of glucose. Unexpectedly, we discovered that G. stearothermophilus T-1 can also utilize lactose and galactosyl-glycerol via the cellobiose-PTS system together with a bifunctional 6-phospho-β-gal/glucosidase, Gan1D. Growth curves of strain T-1 growing in the presence of cellobiose, with either lactose or galactosyl-glycerol, revealed initially logarithmic growth on cellobiose and then linear growth supported by the additional sugars. We conclude that Gan1D allows the cell to utilize residual galactose-containing disaccharides, taking advantage of the promiscuity of the cellobiose-PTS system.

scholarly journals 2-Deoxy-d-galactose, a substrate for the galactose-transport system of Escherichia coli

1977 ◽  
Vol 168 (1) ◽  
pp. 15-22 ◽  
Author(s):  
P J F Henderson ◽  
R A Giddens
Keyword(s):  
Escherichia Coli ◽  
Transport System ◽  
Transport Systems ◽  
Mutual Inhibition ◽  
E Coli ◽  
System 2 ◽  
Galactose Transport ◽  
Proton Uptake ◽  
Inhibition Studies ◽  
Lactose Transport

The following observations showed that 2-deoxy-D-galactose is a useful tool for the isolation and elucidation of the activity of one system for galactose uptake into Escherichia coli. 1. 2-Deoxygalactose, which is not a substrate for growth of E. coli, was transported into strains of the organism induced for galactose transport. 2. By using appropriate mutants it was shown that 2-deoxygalactose is a much better substrate for the galactose-transport system than for the methyl galactoside-transport system. This was confirmed by the results of mutual inhibition studies with substrates of each transport system. 3. The glucose-, arabinose- or lactose-transport systems did not effect significant transport of 2-deoxygalactose. 4. Like other substrates of the galactose-transport system, 2-deoxygalactose promoted effective proton uptake into de-energized suspensions of appropriate E. coli strains. 5. The S183 series of E. coli mutants were found to contain a constitutive galactose-transport system, if 2-deoxygalactose transport is used as one criterion for such activity.

scholarly journals Transport of galactose, glucose and their molecular analogues by Escherichia coli K12

1977 ◽  
Vol 162 (2) ◽  
pp. 309-320 ◽  
Author(s):  
P J F Henderson ◽  
R A Giddens ◽  
M C Jones-Mortimer
Keyword(s):  
Escherichia Coli ◽  
Transport System ◽  
Proton Transport ◽  
Regulatory Mechanism ◽  
Transport Systems ◽  
British Library ◽  
Transport Activity ◽  
Phosphotransferase System ◽  
Escherichia Coli K12 ◽  
Specific Transport System

1. Strains of Escherichia coli K12 were made that are unable to assimilate glucose by the phosphotransferase system, since they lack the glucose-specific components specified by the genes ptsG and ptsM. 2. Derivative organisms lacking the methyl galactoside or galactose-specific transport system were examined for their ability to transport galactose, d-fucose, methyl beta-D-galactoside, glucose, 2-deoxy-D-glucose and methyl alpha-D-glucoside. 3. Galactose, glucose and to a lesser extent fucose are substrates for both transport systems. 4. 2-Deoxyglucose is transported on the galactose-specific but not the methyl galactoside system. 5. The ability of sugars to elicit anaerobic proton transport is associated with the galactose-specific, but not with the methyl galactoside transport activity. Hence a chemiosmotic mechanism of energization is likely to apply to the former but not to the latter. Alternatively the methyl galactoside system may be switched off under certain conditions, which would indicate a novel regulatory mechanism. 6. Details of the procedure for the derivation of strains may be obtained from the authors, and have been deposited as Supplementary Publication SUP 50074 (8 pages at the) British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1977), 161,1.

scholarly journals Role of Periplasmic Trehalase in Uptake of Trehalose by the Thermophilic Bacterium Rhodothermus marinus

2008 ◽  
Vol 190 (6) ◽  
pp. 1871-1878
Author(s):  
Carla D. Jorge ◽  
Luís L. Fonseca ◽  
Winfried Boos ◽  
Helena Santos
Keyword(s):  
Kinetic Parameters ◽  
Glucose Transport ◽  
Transport System ◽  
Glucose Transporter ◽  
Thermophilic Bacterium ◽  
Transport Systems ◽  
Uptake System ◽  
Rhodothermus Marinus ◽  
Whole Cells ◽  
Physiological Relevance

ABSTRACT Trehalose uptake at 65°C in Rhodothermus marinus was characterized. The profile of trehalose uptake as a function of concentration showed two distinct types of saturation kinetics, and the analysis of the data was complicated by the activity of a periplasmic trehalase. The kinetic parameters of this enzyme determined in whole cells were as follows: Km = 156 ± 11 μM and V max = 21.2 ± 0.4 nmol/min/mg of total protein. Therefore, trehalose could be acted upon by this periplasmic activity, yielding glucose that subsequently entered the cell via the glucose uptake system, which was also characterized. To distinguish the several contributions in this intricate system, a mathematical model was developed that took into account the experimental kinetic parameters for trehalase, trehalose transport, glucose transport, competition data with trehalose, glucose, and palatinose, and measurements of glucose diffusion out of the periplasm. It was concluded that R. marinus has distinct transport systems for trehalose and glucose; moreover, the experimental data fit perfectly with a model considering a high-affinity, low-capacity transport system for trehalose (Km = 0.11 ± 0.03 μM and V max = 0.39 ± 0.02 nmol/min/mg of protein) and a glucose transporter with moderate affinity and capacity (Km = 46 ± 3 μM and V max = 48 ± 1 nmol/min/mg of protein). The contribution of the trehalose transporter is important only in trehalose-poor environments (trehalose concentrations up to 6 μM); at higher concentrations trehalose is assimilated primarily via trehalase and the glucose transport system. Trehalose uptake was constitutive, but the activity decreased 60% in response to osmotic stress. The nature of the trehalose transporter and the physiological relevance of these findings are discussed.

scholarly journals Carbohydrate Utilization Patterns for the Extremely Thermophilic Bacterium Caldicellulosiruptor saccharolyticus Reveal Broad Growth Substrate Preferences

2009 ◽  
Vol 75 (24) ◽  
pp. 7718-7724 ◽  
Author(s):  
Amy L. VanFossen ◽  
Marcel R. A. Verhaart ◽  
Servé M. W. Kengen ◽  
Robert M. Kelly
Keyword(s):  
Transcriptional Response ◽  
Thermophilic Bacterium ◽  
Open Reading Frames ◽  
Transport Systems ◽  
Response Analysis ◽  
Growth Substrate ◽  
Phosphotransferase System ◽  
Carbohydrate Utilization ◽  
Caldicellulosiruptor Saccharolyticus ◽  
Substrate Preferences

ABSTRACT Coutilization of hexoses and pentoses derived from lignocellulose is an attractive trait in microorganisms considered for consolidated biomass processing to biofuels. This issue was examined for the H2-producing, extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus growing on individual monosaccharides (arabinose, fructose, galactose, glucose, mannose, and xylose), mixtures of these sugars, as well as on xylan and xylogluco-oligosacchrides. C. saccharolyticus grew at approximately the same rate (t d, ∼95 min) and to the same final cell density (1 × 108 to 3 × 108 cells/ml) on all sugars and sugar mixtures tested. In the monosaccharide mixture, although simultaneous consumption of all monosaccharides was observed, not all were utilized to the same extent (fructose > xylose/arabinose > mannose/glucose/galactose). Transcriptome contrasts for monosaccharide growth revealed minimal changes in some cases (e.g., 32 open reading frames [ORFs] changed ≥2-fold for glucose versus galactose), while substantial changes occurred for cases involving mannose (e.g., 353 ORFs changed ≥2-fold for glucose versus mannose). Evidence for catabolite repression was not noted for either growth on multisugar mixtures or the corresponding transcriptomes. Based on the whole-genome transcriptional response analysis and comparative genomics, carbohydrate specificities for transport systems could be proposed for most of the 24 putative carbohydrate ATP-binding cassette transporters and single phosphotransferase system identified in C. saccharolyticus. Although most transporter genes responded to individual monosaccharides and polysaccharides, the genes Csac_0692 to Csac_0694 were upregulated only in the monosaccharide mixture. The results presented here affirm the broad growth substrate preferences of C. saccharolyticus on carbohydrates representative of lignocellulosic biomass and suggest that this bacterium holds promise for biofuel applications.

scholarly journals The High-Affinity Maltose/Trehalose ABC Transporter in the Extremely Thermophilic Bacterium Thermus thermophilus HB27 Also Recognizes Sucrose and Palatinose

2005 ◽  
Vol 187 (4) ◽  
pp. 1210-1218 ◽  
Author(s):  
Zélia Silva ◽  
Maria-Manuel Sampaio ◽  
Anke Henne ◽  
Alex Böhm ◽  
Ruben Gutzat ◽  
...  
Keyword(s):  
Abc Transporter ◽  
Transport System ◽  
Equilibrium Dialysis ◽  
Thermus Thermophilus ◽  
Gene Clusters ◽  
Thermophilic Bacterium ◽  
Binding Activity ◽  
Transport Systems ◽  
Growth Defect ◽  
High Affinity

ABSTRACT We have studied the transport of trehalose and maltose in the thernophilic bacterium Thermus thermophilus HB27, which grows optimally in the range of 70 to 75°C. The Km values at 70°C were 109 nM for trehalose and 114 nM for maltose; also, a high Km (424 nM) was found for the uptake of sucrose. Competition studies showed that a single transporter recognizes trehalose, maltose, and sucrose, while d-galactose, d-fucose, l-rhamnose, l-arabinose, and d-mannose were not competitive inhibitors. In the recently published genome of T. thermophilus HB27, two gene clusters designated malEFG1 (TTC1627 to -1629) and malEFG2 (TTC1288 to -1286) and two monocistronic genes designated malK1 (TTC0211) and malK2 (TTC0611) are annotated as trehalose/maltose and maltose/maltodextrin transport systems, respectively. To find out whether any of these systems is responsible for the transport of trehalose, the malE1 and malE2 genes, lacking the sequence encoding the signal peptides, were expressed in Escherichia coli. The binding activity of pure recombinant proteins was analyzed by equilibrium dialysis. MalE1 was able to bind maltose, trehalose, and sucrose but not glucose or maltotetraose (Kd values of 103, 67, and 401 nM, respectively). Mutants with disruptions in either malF1 or malK1 were unable to grow on maltose, trehalose, sucrose, or palatinose, whereas mutants with disruption in malK2 or malF2 showed no growth defect on any of these sugars. Therefore, malEFG1 encodes the binding protein and the two transmembrane subunits of the trehalose/maltose/sucrose/palatinose ABC transporter, and malK1 encodes the ATP-binding subunit of this transporter. Despite the presence of an efficient transporter for trehalose, this compound was not used by HB27 for osmoprotection. MalE1 and MalE2 exhibited extremely high thermal stability: melting temperatures of 90°C for MalE1 and 105°C for MalE2 in the presence of 2.3 M guanidinium chloride. The latter protein did not bind any of the sugars examined and is not implicated in a maltose/maltodextrin transport system. This work demonstrates that malEFG1 and malK1 constitute the high-affinity ABC transport system of T. thermophilus HB27 for trehalose, maltose, sucrose, and palatinose.

scholarly journals Genome Sequence of Geobacillus stearothermophilus DSM 458, an Antimicrobial-Producing Thermophilic Bacterium, Isolated from a Sugar Beet Factory

2017 ◽  
Vol 5 (43) ◽  
Author(s):  
Kevin Egan ◽  
Philip Kelleher ◽  
Des Field ◽  
Mary C. Rea ◽  
R. Paul Ross ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Genome Sequence ◽  
In Silico ◽  
Thermophilic Bacterium ◽  
Full Genome Sequence ◽  
Full Genome ◽  
Geobacillus Stearothermophilus ◽  
Biosynthetic Genes ◽  
Content Type

ABSTRACT This paper reports the full genome sequence of the antimicrobial-producing bacterium Geobacillus stearothermophilus DSM 458, isolated in a sugar beet factory in Austria. In silico analysis reveals the presence of a number of novel bacteriocin biosynthetic genes.

Heave Induced Internal Flow Fluctuations in Vertical Transport Systems for Deep Ocean Mining

2014 ◽  
Author(s):  
Stephan D. A. Hannot ◽  
Jort M. van Wijk
Keyword(s):  
Transport System ◽  
Internal Flow ◽  
Deep Ocean ◽  
Vertical Transport ◽  
Transport Systems ◽  
Ship Motions ◽  
Centrifugal Pumps ◽  
Mining System ◽  
Pump System ◽  
Ocean Mining

Deep ocean mining systems will have to operate often in harsh weather conditions with heavy sea states. A typical mining system consists of a Mining Support Vessel (MSV) with a Vertical Transport System (VTS) attached to it. The transport system is a pump pipeline system using centrifugal pumps. The heave motions of the ship are transferred to the pump system due to the riser-ship coupling. Ship motions thus will have a significant influence on the internal flow in the VTS. In this paper, the influence of heave motions on the internal flow in the VTS for a typical mining system for Seafloor Massive Sulfide (SMS) deposits in Papua New Guinea is analyzed. Data on the wave climate in the PNG region is used to compute the ship motions of a coupled MSV-VTS. The ship motions then are translated into forces acting on the internal flow in order to compute fluctuations in the internal flow. In this way, the workability of the mining system with respect to the system’s production can be assessed. Based on a detailed analysis of the internal flow in relation to ship motions, the relevance of a coupled analysis for the design of VTS is made clear. This paper provides a method for performing such analyses.

scholarly journals Trk2 Potassium Transport System in Streptococcus mutans and Its Role in Potassium Homeostasis, Biofilm Formation, and Stress Tolerance

2016 ◽  
Vol 198 (7) ◽  
pp. 1087-1100 ◽  
Author(s):  
Gursonika Binepal ◽  
Kamal Gill ◽  
Paula Crowley ◽  
Martha Cordova ◽  
L. Jeannine Brady ◽  
...  
Keyword(s):  
Stress Tolerance ◽  
Streptococcus Mutans ◽  
Transport System ◽  
Biofilm Formation ◽  
Cellular Response ◽  
Pathogenic Bacteria ◽  
Transport Systems ◽  
Content Type ◽  
Growth And Survival ◽  
Dental Biofilm

ABSTRACTPotassium (K+) is the most abundant cation in the fluids of dental biofilm. The biochemical and biophysical functions of K+and a variety of K+transport systems have been studied for most pathogenic bacteria but not for oral pathogens. In this study, we establish the modes of K+acquisition inStreptococcus mutansand the importance of K+homeostasis for its virulence attributes. TheS. mutansgenome harbors four putative K+transport systems that included two Trk-like transporters (designated Trk1 and Trk2), one glutamate/K+cotransporter (GlnQHMP), and a channel-like K+transport system (Kch). Mutants lacking Trk2 had significantly impaired growth, acidogenicity, aciduricity, and biofilm formation. [K+] less than 5 mM eliminated biofilm formation inS. mutans. The functionality of the Trk2 system was confirmed by complementing anEscherichia coliTK2420 mutant strain, which resulted in significant K+accumulation, improved growth, and survival under stress. Taken together, these results suggest that Trk2 is the main facet of the K+-dependent cellular response ofS. mutansto environment stresses.IMPORTANCEBiofilm formation and stress tolerance are important virulence properties of caries-causingStreptococcus mutans. To limit these properties of this bacterium, it is imperative to understand its survival mechanisms. Potassium is the most abundant cation in dental plaque, the natural environment ofS. mutans. K+is known to function in stress tolerance, and bacteria have specialized mechanisms for its uptake. However, there are no reports to identify or characterize specific K+transporters inS. mutans. We identified the most important system for K+homeostasis and its role in the biofilm formation, stress tolerance, and growth. We also show the requirement of environmental K+for the activity of biofilm-forming enzymes, which explains why such high levels of K+would favor biofilm formation.

scholarly journals Inactivation of the galactose transport system inSaccharomyces cerevisiae

FEBS Letters ◽  
1986 ◽  
Vol 207 (2) ◽  
pp. 258-261 ◽  
Author(s):  
Carmen DeJuan ◽  
Rosario Lagunas
Keyword(s):  
Transport System ◽  
Galactose Transport

scholarly journals Regarding some aspects of economic feasibility for the transport integrated use in favor of environmental security

2018 ◽  
Vol 170 ◽  
pp. 05001 ◽  
Author(s):  
Ruben Kazaryan
Keyword(s):  
National Security ◽  
Transport System ◽  
Developed Countries ◽  
Climatic Conditions ◽  
Environmental Security ◽  
Economic Feasibility ◽  
Transport Systems ◽  
Domestic Water ◽  
European Area ◽  
High Dynamics

The transport system in Russia and the transport support issues of national security differ completely from the Occident and other developed countries due to geostrategic and natural climatic conditions of our country. The integrated transport system, where in a sufficiently coherent manner used are the roads and railways, domestic water and air ways, pipeline transport, is more developed in the European area of the country. However there is no assessment of economic feasibility for the integrated transport lines. Firstly, assessment criteria for the transport support efficiency in favor of environmental security; secondly, mathematical economic models of various transport types integrated use in favor of environmental security; thirdly, economic feasibility methods for transport integrated use in favor of environmental security, that unavoidably leads to unreasonable transport operations using the existing rail and water ways at little distances, and increases their base cost by several times versus the transport by road. High dynamics of transport support for economic, demographic and environmental security within a market economy require the development of approach for the transport integrated use in favor of Russia national security. This article describes some practical recommendations for transport systems control bodies.

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