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Articles 151 - 180 of 192

Full-Text Articles in Biological Engineering

Determination Of Master Compliance Curve For Extruded Semolina Pasta, Laura E. Emery, Martin Okos Oct 2013

Determination Of Master Compliance Curve For Extruded Semolina Pasta, Laura E. Emery, Martin Okos

The Summer Undergraduate Research Fellowship (SURF) Symposium

The dependence of the rate of relaxation of semolina pasta on moisture content and temperature and how it affects shrinkage during drying has not yet been determined. The purpose of this research was to develop an equation that relates moisture content and temperature in order to obtain a master curve for creep of the product. When found, this equation could help to optimize the drying process and increase the quality of the final pasta product. Semolina flour mixed with water and propionic acid to create a 35% wet basis product was extruded on a C.W. Brabender 2523 to obtain a …


Artificial Yeast Polarization Controlled By Chemical Gradient, James K. Nolan, Bernard Tao Oct 2013

Artificial Yeast Polarization Controlled By Chemical Gradient, James K. Nolan, Bernard Tao

The Summer Undergraduate Research Fellowship (SURF) Symposium

Engineering synthetic multicellular systems will lead to new synthetic biology technological platforms, inform developmental biology through recapitulation of natural systems and possibly unveil novel morphologies with practical applications not before reached throughout natural history (Maharbiz, 2012). Creating an exogenous molecular circuit that will polarize unicellular cells into “apical” and “basal” domains relative to a substrate plane would fulfill a missing component towards fully multicellular synthetic cellular communities (Maharbiz, 2012). To this end, a PIP3 polarization network previously designed by Chau and associates (Chau, Walter, Gerardin, Tang, Lim 2012) was coupled to the specific activation by niacin of a recombinant …


Life Support: Long Term Storage Of Solid Waste In An Enclosed Membrane System, Emmy O. Trieu, Michael Flynn, Rocco Mancinelli Aug 2013

Life Support: Long Term Storage Of Solid Waste In An Enclosed Membrane System, Emmy O. Trieu, Michael Flynn, Rocco Mancinelli

STAR Program Research Presentations

In deep space missions, maintaining life support is of the utmost priority. In such a closed system, human waste must be stored and treated. Simulated feces (ersatz) will be inoculated with microbes representing normal fecal flora, mixed with urine brine and shredded refuse of products typically used in space missions. Composting methods often use alternating layers of waste with scraps of carboniferous materials (finely shredded refuse). By preparing membrane bags with a homogenized ersatz and carboniferous refuse mixture and membrane bags with alternating layers of ersatz and carboniferous refuse, it may be possible to monitor anaerobic thermophillic digestion of the …


Controlling Bacterial Persister Cells With Low Level Electric Currents, Henry Lars Peterson May 2013

Controlling Bacterial Persister Cells With Low Level Electric Currents, Henry Lars Peterson

Renée Crown University Honors Thesis Projects - All

Bacterial persister cells present a growing concern as they inherit the ability to tolerate high concentrations of antibiotics and repopulate after an antibiotic treatment leading to chronic diseases. Pseudomonas aeruginosa causes many human infections including skin infections and those associated with burn injuries, and implanted medical devices, and are associated with Cystic Fibrosis. Recently, the Ren Lab developed a novel approach to eliminate persister cells of P. aeruginosa, including those in biofilms, using low level electric currents. To evaluate the safety of this method and to better understand how the underlying elements, this study focused on the cytotoxicity of …


Atypical Glycolysis In Clostridium Thermocellum, Jilai Zhou, Daniel G. Olson, D. Aaron Argyros, Yu Deng, Walter M. Van Gulik, Johannes P. Van Dijken, Lee R. Lynd Feb 2013

Atypical Glycolysis In Clostridium Thermocellum, Jilai Zhou, Daniel G. Olson, D. Aaron Argyros, Yu Deng, Walter M. Van Gulik, Johannes P. Van Dijken, Lee R. Lynd

Dartmouth Scholarship

Cofactor specificities of glycolytic enzymes in Clostridium thermocellum were studied with cellobiose-grown cells from batch cultures. Intracellular glucose was phosphorylated by glucokinase using GTP rather than ATP. Although phosphofructokinase typically uses ATP as a phosphoryl donor, we found only pyrophosphate (PPi)-linked activity. Phosphoglycerate kinase used both GDP and ADP as phosphoryl acceptors. In agreement with the absence of a pyruvate kinase sequence in the C. thermocellum genome, no activity of this enzyme could be detected. Also, the annotated pyruvate phosphate dikinase (ppdk) is not crucial for the generation of pyruvate from phosphoenolpyruvate (PEP), as deletion of the ppdk gene did …


The Effects Of Hydrostatic Pressure On Early Endothelial Tubulogenic Processes, Ryan M. Underwood Jan 2013

The Effects Of Hydrostatic Pressure On Early Endothelial Tubulogenic Processes, Ryan M. Underwood

Theses and Dissertations--Biomedical Engineering

The effects of mechanical forces on endothelial cell function and behavior are well documented, but have not been fully characterized. Specifically, fluid pressure has been shown to elicit physical and chemical responses known to be involved in the initiation and progression of endothelial cell-mediated vascularization. Central to the process of vascularization is the formation of tube-like structures. This process—tubulogenesis—is essential to both the physiological and pathological growth of tissues. Given the known effects of pressure on endothelial cells and its ubiquitous presence in the vasculature, we investigated pressure as a magnitude-dependent parameter for the regulation of endothelial tubulogenic activity. To …


Noninvasive Near-Infrared Diffuse Optical Monitoring Of Cerebral Hemodynamics And Autoregulation, Ran Cheng Jan 2013

Noninvasive Near-Infrared Diffuse Optical Monitoring Of Cerebral Hemodynamics And Autoregulation, Ran Cheng

Theses and Dissertations--Biomedical Engineering

Many cerebral diseases are associated with abnormal cerebral hemodynamics and impaired cerebral autoregulation (CA). CA is a mechanism to maintain cerebral blood flow (CBF) stable when mean arterial pressure (MAP) fluctuates. Evaluating these abnormalities requires direct measurements of cerebral hemodynamics and MAP. Several near-infrared diffuse optical instruments have been developed in our laboratory for hemodynamic measurements including near-infrared spectroscopy (NIRS), diffuse correlation spectroscopy (DCS), hybrid NIRS/DCS, and dual-wavelength DCS flow-oximeter. We utilized these noninvasive technologies to quantify CBF and cerebral oxygenation in different populations under different physiological conditions/manipulations. A commercial finger plethysmograph was used to continuously monitor MAP. For investigating …


Bacillus Thuringiensis: Transgenic Crops, Julie A. Peterson, John J. Obrycki, James D. Harwood Jan 2013

Bacillus Thuringiensis: Transgenic Crops, Julie A. Peterson, John J. Obrycki, James D. Harwood

Department of Entomology: Faculty Publications

Bacillus thuringiensis (Bt) crops, genetically modified to express insecticidal toxins that target key pests of corn, cotton, rice, potato, and other crops, have been rapidly adopted and have become dominant fixtures in agroecosystems throughout the world. Due to the constitutive nature of Bt toxin expression, insecticidal proteins may be found in nearly all plant tissues, presenting multiple sources for Bt toxins to enter the environment, thus creating complex direct and indirect pathways for non-target organisms to be exposed to insecticidal proteins. The environmental impacts of Bt crops have been widely debated, although both benefits and risks do exist. Benefits of …


Characterization Of Xylan Utilization And Discovery Of A New Endoxylanase In Thermoanaerobacterium Saccharolyticum Through Targeted Gene Deletions, Kara K. Podkaminer, Adam M. Guss, Heather L. Trajano, David A. Hogsett, Lee R. Lynd Sep 2012

Characterization Of Xylan Utilization And Discovery Of A New Endoxylanase In Thermoanaerobacterium Saccharolyticum Through Targeted Gene Deletions, Kara K. Podkaminer, Adam M. Guss, Heather L. Trajano, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

The economical production of fuels and commodity chemicals from lignocellulose requires the utilization of both the cellulose and hemicellulose fractions. Xylanase enzymes allow greater utilization of hemicellulose while also increasing cellulose hydrolysis. Recent metabolic engineering efforts have resulted in a strain of Thermoanaerobacterium saccharolyticum that can convert C5 and C6 sugars, as well as insoluble xylan, into ethanol at high yield. To better understand the process of xylan solubilization in this organism, a series of targeted deletions were constructed in the homoethanologenic T. saccharolyticum strain M0355 to characterize xylan hydrolysis and xylose utilization in this organism. While the deletion of …


Ethanol And Anaerobic Conditions Reversibly Inhibit Commercial Cellulase Activity In Thermophilic Simultaneous Saccharification And Fermentation (Tssf), Kara K. Podkaminer, William R. Kenealy, Christopher D. Herring, David A. Hogsett, Lee R. Lynd Jun 2012

Ethanol And Anaerobic Conditions Reversibly Inhibit Commercial Cellulase Activity In Thermophilic Simultaneous Saccharification And Fermentation (Tssf), Kara K. Podkaminer, William R. Kenealy, Christopher D. Herring, David A. Hogsett, Lee R. Lynd

Dartmouth Scholarship

A previously developed mathematical model of low solids thermophilic simultaneous saccharification and fermentation (tSSF) with Avicel was unable to predict performance at high solids using a commercial cellulase preparation (Spezyme CP) and the high ethanol yield Thermoanaerobacterium saccharolyticum strain ALK2. The observed hydrolysis proceeded more slowly than predicted at solids concentrations greater than 50 g/L Avicel. Factors responsible for this inaccuracy were investigated in this study.


Enhanced Hydrocarbon Biodegradation Using Bioaugmentation With Biowish™-Aqua Fog, Michael Robert Lehrer Jun 2012

Enhanced Hydrocarbon Biodegradation Using Bioaugmentation With Biowish™-Aqua Fog, Michael Robert Lehrer

Master's Theses

This study was done to determine the effectiveness of a commercially available bioaugmentation product, BiOWiSHTM-Aqua FOG, for remediating petroleum-contaminated sandy soil. Biodegradation enhancement by BiOWiSHTM-Aqua FOG was evaluated in laboratory microcosms by directly measuring total petroleum hydrocarbon (TPH) and indirectly using respirometry. Attempts were made to enrich hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG, and the resulting enrichment cultures were screened using respirometry as well. Potential hydrocarbon-degrading bacteria in BiOWiSHTM-Aqua FOG were isolated. Experiments were performed at bench-scale using microcosm bottles containing sand contaminated with either motor oil or No. 2 diesel fuel. The …


Development Of A Protocol To Measure Gene Expression In The Mouse Tibia, Daniel Hoover Jun 2012

Development Of A Protocol To Measure Gene Expression In The Mouse Tibia, Daniel Hoover

Biomedical Engineering

Numerous molecular factors active in bone tissue direct fracture repair and remodeling which can be altered by disease conditions such as Peripheral Arterial Disease (PAD) and Osteoporosis. Methods of molecular biology are commonly applied to investigate the expression and role of these molecular factors. This project presents a robust three-step protocol for examining gene expression in the mouse tibia. The protocol begins with isolating RNA from a flash frozen tibia sample. The isolated RNA is reverse transcribed into cDNA. Finally, PCR is performed to indentify expressed genes. Establishing this protocol will allow further research into the mechanisms of bone remodeling …


Computational Design And Characterization Of A Temperature-Sensitive Plasmid Replicon For Gram Positive Thermophiles, Daniel G. Olson, Lee R. Lynd May 2012

Computational Design And Characterization Of A Temperature-Sensitive Plasmid Replicon For Gram Positive Thermophiles, Daniel G. Olson, Lee R. Lynd

Dartmouth Scholarship

Temperature-sensitive (Ts) plasmids are useful tools for genetic engineering, but there are currently none compatible with the gram positive, thermophilic, obligate anaerobe, Clostridium thermocellum. Traditional mutagenesis techniques yield Ts mutants at a low frequency, and therefore requires the development of high-throughput screening protocols, which are also not available for this organism. Recently there has been progress in the development of computer algorithms which can predict Ts mutations. Most plasmids currently used for genetic modification of C. thermocellum are based on the replicon of plasmid pNW33N, which replicates using the RepB replication protein. To address this problem, we set out …


Design And Implementation Of A Shoulder Simulator, Chelsea Lynn Kowal May 2012

Design And Implementation Of A Shoulder Simulator, Chelsea Lynn Kowal

Renée Crown University Honors Thesis Projects - All

The main objective of this thesis project is to build a shoulder simulator to be able to understand the mechanics of the shoulder, specifically the rotator cuff and deltoid muscles. The shoulder simulator should be able to mimic the important motions of the human shoulder by pulling on tendons of cadaver shoulders and moving the shoulder in various specified motions. In my thesis project, I worked on creating a shoulder simulator that would control the rotator cuff muscles along with the three deltoid muscles. In order to do this, I needed to design and test the shoulder simulator using cadaver …


A Thousand Tiny Pieces: The Federal Circuit’S Fractured Myriad Ruling, Lessons To Be Learned, And The Way Forward, Jonathan R. K. Stroud Jan 2012

A Thousand Tiny Pieces: The Federal Circuit’S Fractured Myriad Ruling, Lessons To Be Learned, And The Way Forward, Jonathan R. K. Stroud

IP Theory

No abstract provided.


Complete Genome Sequence Of Clostridium Clariflavum Dsm 19732, Javier A. Izquierdo, Lynne Goodwin, Karen W. Davenport, Hazuki Teshima Jan 2012

Complete Genome Sequence Of Clostridium Clariflavum Dsm 19732, Javier A. Izquierdo, Lynne Goodwin, Karen W. Davenport, Hazuki Teshima

Dartmouth Scholarship

Clostridium clariflavum is a Cluster III Clostridium within the family Clostridiaceae isolated from thermophilic anaerobic sludge (Shiratori et al, 2009). This species is of interest because of its similarity to the model cellulolytic organism Clostridium thermocellum and for the ability of environmental isolates to break down cellulose and hemicellulose. Here we describe features of the 4,897,678 bp long genome and its annotation, consisting of 4,131 protein-coding and 98 RNA genes, for the type strain DSM 19732.


An Investigation Of Reflective Mulches For Use Over Capillary Mat Systems For Winter-Time Greenhouse Strawberry Production, George E. Meyer, Ellen T. Paparozzi, Elizabeth Walter-Shea, Erin E. Blankenship, Stacy A. Adams Jan 2012

An Investigation Of Reflective Mulches For Use Over Capillary Mat Systems For Winter-Time Greenhouse Strawberry Production, George E. Meyer, Ellen T. Paparozzi, Elizabeth Walter-Shea, Erin E. Blankenship, Stacy A. Adams

Department of Agronomy and Horticulture: Faculty Publications

Photosynthethically active radiation (PAR) is a principle environmental variable used by horticultural specialists, agronomists and ecosystem modelers to characterize the quantity and quality of light conducive to plant growth and development. Spatial distribution of PAR in a greenhouse can be quite variable and diffuse throughout the day time photoperiod, especially at low sun angles in northern regions of the United States. Four colors of reflective plastic mulches (white, red, olive, and black) were evaluated for winter-time strawberry (Fragaria × ananassa Duch.) production based on their reflectance and transmittance properties in a double-polyethylene, plastic-glazed Quonset greenhouse inNebraska. The spectral properties …


Enhanced Microbial Utilization Of Recalcitrant Cellulose By An Ex Vivo Cellulosome-Microbe Complex, Chun You, Xiao-Zhou Zhang, Noppadon Sathitsuksanoh, Lee R. Lynd Dec 2011

Enhanced Microbial Utilization Of Recalcitrant Cellulose By An Ex Vivo Cellulosome-Microbe Complex, Chun You, Xiao-Zhou Zhang, Noppadon Sathitsuksanoh, Lee R. Lynd

Dartmouth Scholarship

A cellulosome-microbe complex was assembled ex vivo on the surface of Bacillus subtilis displaying a miniscaffoldin that can bind with three dockerin-containing cellulase components: the endoglucanase Cel5, the processive endoglucanase Cel9, and the cellobiohydrolase Cel48. The hydrolysis performances of the synthetic cellulosome bound to living cells, the synthetic cellulosome, a noncomplexed cellulase mixture with the same catalytic components, and a commercial fungal enzyme mixture were investigated on low-accessibility recalcitrant Avicel and high accessibility regenerated amorphous cellulose (RAC). The cellbound cellulosome exhibited 4.5- and 2.3-fold-higher hydrolysis ability than cell-free cellulosome on Avicel and RAC, respectively. The cellulosome-microbe synergy was not completely …


Design And Fabrication Of Nanofluidic Systems For Biomolecule Characterizations, Orain Ansel Hibbert Dec 2011

Design And Fabrication Of Nanofluidic Systems For Biomolecule Characterizations, Orain Ansel Hibbert

Graduate Theses and Dissertations

Nanofluidic channel systems were designed and fabricated by combining MEMS microfabrication with AFM nanolithography. In the fabrication process flow, photolithography was first utilized to pattern microfluidic channels and reservoirs on a 4" Pyrex substrate. Subsequently, atomic force microscopy (AFM) based nanolithography was used to mechanically fabricate nanochannels to connect the microreservoirs which formed the inlet and outlet of the nanofluidic system. A Tap190 Diamond-Like Carbon (DLC) AFM tip with a force constant of 48 N/m and a radius of less than 15 nm was used as the nanolithography tool. The resultant nanochannel ranges from 20 to 80 µm in length …


Comparison Of Four Chip-Seq Analytical Algorithms Using Rice Endosperm H3k27 Trimethylation Profiling Data., Brandon M. Malone, Feng Tan, Susan M. Bridges, Zhaohua Peng Sep 2011

Comparison Of Four Chip-Seq Analytical Algorithms Using Rice Endosperm H3k27 Trimethylation Profiling Data., Brandon M. Malone, Feng Tan, Susan M. Bridges, Zhaohua Peng

BCoE Publications

Chromatin immunoprecipitation coupled with high throughput DNA Sequencing (ChIP-Seq) has emerged as a powerful tool for genome wide profiling of the binding sites of proteins associated with DNA such as histones and transcription factors. However, no peak calling program has gained consensus acceptance by the scientific community as the preferred tool for ChIP-Seq data analysis. Analyzing the large data sets generated by ChIP-Seq studies remains highly challenging for most molecular biology laboratories.Here we profile H3K27me3 enrichment sites in rice young endosperm using the ChIP-Seq approach and analyze the data using four peak calling algorithms (FindPeaks, PeakSeq, USeq, and MACS). Comparison …


Mutant Alcohol Dehydrogenase Leads To Improved Ethanol Tolerance In Clostridium Thermocellum, Steven D. Brown, Adam M. Guss, Tatiana V. Karpinets, Jerry M. Parks Aug 2011

Mutant Alcohol Dehydrogenase Leads To Improved Ethanol Tolerance In Clostridium Thermocellum, Steven D. Brown, Adam M. Guss, Tatiana V. Karpinets, Jerry M. Parks

Dartmouth Scholarship

Clostridium thermocellum is a thermophilic, obligately anaerobic, Gram-positive bacterium that is a candidate microorganism for converting cellulosic biomass into ethanol through consolidated bioprocessing. Ethanol intolerance is an important metric in terms of process economics, and tolerance has often been described as a complex and likely multigenic trait for which complex gene interactions come into play. Here, we resequence the genome of an ethanol-tolerant mutant, show that the tolerant phenotype is primarily due to a mutated bifunctional acetaldehyde-CoA/alcohol dehydrogenase gene (adhE), hypothesize based on structural analysis that cofactor specificity may be affected, and confirm this hypothesis using enzyme assays. …


Forward Osmosis Bag: Ground Testing The Prospect Of Using Alternative Water Sources For Drinking Water, Michaela Marie Brant, Ariella Yendler, Mona Hammoudeh, Michael T. Flynn Aug 2011

Forward Osmosis Bag: Ground Testing The Prospect Of Using Alternative Water Sources For Drinking Water, Michaela Marie Brant, Ariella Yendler, Mona Hammoudeh, Michael T. Flynn

STAR Program Research Presentations

On the STS-135 Space Shuttle mission, to be launched July 8, 2011, a forward osmosis bag (FOB) study will be conducted. At NASA Ames this summer, the ground truth testing is being conducted for results comparisons. The FOB technology is derived from a commercial product, the X-Pack water filter. Forward osmosis operates by utilizing an established concentration gradient across a semi-permeable membrane to move water molecules from one side of the membrane to the other. This concept is exploited to harvest drinking water from grey water sources such as urine, sea water, or vehicle water. In this experiment, potassium chloride …


Identifying Changes In Climatic Trends And The Fingerprints Of Landuse And Landcover Changes In The High Plains Of The Usa, Denis Mutiibwa Jul 2011

Identifying Changes In Climatic Trends And The Fingerprints Of Landuse And Landcover Changes In The High Plains Of The Usa, Denis Mutiibwa

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

Human activities such as conversion of natural ecosystem to croplands and urban-centers, deforestation and afforestation impact biophysical properties of land surface such as albedo, energy balance, and surface roughness. Alterations in these properties affect the heat and moisture exchanges between the land surface and atmospheric boundary layer. The objectives of this research were; (i) to quantitatively identify the High plains’ regional climate change in temperatures over the period 1895 to 2006, (ii) detect the signatures of anthropogenic forcing of LULC changes on the regional climate change of the High Plains, and (iii) examine the trends in evolving regional latent heat …


Modeling And Quantitative Analysis Of White Matter Fiber Tracts In Diffusion Tensor Imaging, Xuwei Liang Jan 2011

Modeling And Quantitative Analysis Of White Matter Fiber Tracts In Diffusion Tensor Imaging, Xuwei Liang

University of Kentucky Doctoral Dissertations

Diffusion tensor imaging (DTI) is a structural magnetic resonance imaging (MRI) technique to record incoherent motion of water molecules and has been used to detect micro structural white matter alterations in clinical studies to explore certain brain disorders. A variety of DTI based techniques for detecting brain disorders and facilitating clinical group analysis have been developed in the past few years. However, there are two crucial issues that have great impacts on the performance of those algorithms. One is that brain neural pathways appear in complicated 3D structures which are inappropriate and inaccurate to be approximated by simple 2D structures, …


Analysis Of The Performance Of Peanut Fame In A Single Cylinder Idi Engine And Investigations Of Neat Methyl Ester Influence On Fuel Quality, Jeffery Lewis Jan 2011

Analysis Of The Performance Of Peanut Fame In A Single Cylinder Idi Engine And Investigations Of Neat Methyl Ester Influence On Fuel Quality, Jeffery Lewis

College of Graduate Studies: Theses & Dissertations

Author's abstract: To meet evolving standards set forth by the U.S. EPA and U.S. Energy Policy it is necessary to consider a variety of feedstock for biofuels synthesis and to understand the role individual fatty acids contribute to the overall performance. It is the belief of the authors that excess peanut production would provide an excellent source for high quality biodiesel feedstock, and the complete property and combustion analysis of the fatty acid components will provide insight for future biofuel developments. The preliminary investigations of the project focused on the combustion performance and thermal efficiency of peanut FAME and its …


Modeling Heat Transfer During Cooling Of Ready-To-Eat Meat And Poultry Products Using Three-Dimensional Finite Element Analysis And Web-Based Simulation, Jihan F. Cepeda Jimenez Dec 2010

Modeling Heat Transfer During Cooling Of Ready-To-Eat Meat And Poultry Products Using Three-Dimensional Finite Element Analysis And Web-Based Simulation, Jihan F. Cepeda Jimenez

Department of Agricultural and Biological Systems Engineering: Dissertations, Theses, and Student Research

The meat industry is required to comply with processing performance standards for preventing the growth of foodborne pathogens in products. These performance standards, established by the United States Department of Agriculture - Food Safety and Inspection Service (USDA-FSIS) require a reduction of Salmonella spp (lethality standard) and limit the growth of sporeforming bacteria (stabilization standard) in certain processed meat products. In general, strategies used to comply with these standards are associated with thermal processing. Meat processors have difficulties complying with these performance standards. Moreover, thermal processing deviations are an issue in the meat industry that generate uncertainty regarding the safety …


Deletion Of The Cel48s Cellulase From Clostridium Thermocellum, Daniel G. Olson, Shital A. Tripathi, Richard J. Giannone, Jonathan Lo, Nicky C. Caiazza, David A. Hogsett, Robert L. Hettich, Adam M. Guss, Genia Dubrovsky, Lee R. Lynd Oct 2010

Deletion Of The Cel48s Cellulase From Clostridium Thermocellum, Daniel G. Olson, Shital A. Tripathi, Richard J. Giannone, Jonathan Lo, Nicky C. Caiazza, David A. Hogsett, Robert L. Hettich, Adam M. Guss, Genia Dubrovsky, Lee R. Lynd

Dartmouth Scholarship

Clostridium thermocellum is a thermophilic anaerobic bacterium that rapidly solubilizes cellulose with the aid of a multienzyme cellulosome complex. Creation of knockout mutants for Cel48S (also known as CelS, S(S), and S8), the most abundant cellulosome subunit, was undertaken to gain insight into its role in enzymatic and microbial cellulose solubilization. Cultures of the Cel48S deletion mutant (S mutant) were able to completely solubilize 10 g/L crystalline cellulose. The cellulose hydrolysis rate of the S mutant strain was 60% lower than the parent strain, with the S mutant strain also exhibiting a 40% reduction in cell yield. The cellulosome produced …


Selective Biochlorination Of Hydroxyquinolines By A Flavin-Dependent Halogenase, Fuchao Xu, Amanda Merkley, Dayu Yu, Jixun Zhan Oct 2010

Selective Biochlorination Of Hydroxyquinolines By A Flavin-Dependent Halogenase, Fuchao Xu, Amanda Merkley, Dayu Yu, Jixun Zhan

Biological Engineering Faculty Publications

Rdc2 is a flavin-dependent halogenase from Pochonia chlamydosporia. Through the introduction of a His6-tag to both the N- and C-termini, the isolation yield of Rdc2 from Escherichia coli using Ni-NTA affinity chromatography was increased by three-fold. In vitro reaction of Rdc2 and a flavin reductase (Fre) with seven different hydroxyquinolines revealed that 3-hydroxyquinoline (3), 5-hydroxyquinoline (5), 6-hydroxyquinoline (6), and 7-hydroxyquinoline (7) can be specifically halogenated. These products were prepared by incubating the corresponding substrates with IPTG-induced E. coli BL21(DE3)/Rdc2. They were respectively characterized as 3-hydroxy-4-chloroquinoline (3a), 5-hydroxy-6-chloroquinoline (5a), 5-chloro-6-hydroxyquinoline (6a), and 7-hydroxy-8-chloroquinoline (7a) by NMR and MS …


Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza Aug 2010

Development Of Pyrf-Based Genetic System For Targeted Gene Deletion In Clostridium Thermocellum And Creation Of A Pta Mutant, Shital A. Tripathi, Daniel G. Olson, D. Aaron Argyros, Bethany B. Miller, Trisha F. Barrett, Daniel M. Murphy, Jesse D. Mccool, Anne K. Warner, Vineet B. Rajgarhia, Lee R. Lynd, David A. Hogsett, Nicky C. Caiazza

Dartmouth Scholarship

We report development of a genetic system for making targeted gene knockouts in Clostridium thermocellum, a thermophilic anaerobic bacterium that rapidly solubilizes cellulose. A toxic uracil analog, 5-fluoroorotic acid (5-FOA), was used to select for deletion of the pyrF gene. The ΔpyrF strain is a uracil auxotroph that could be restored to a prototroph via ectopic expression of pyrF from a plasmid, providing a positive genetic selection. Furthermore, 5-FOA was used to select against plasmid-expressed pyrF, creating a negative selection for plasmid loss. This technology was used to delete a gene involved in organic acid production, namely pta, which encodes …


Robust Dynamic Balance Of Ap-1 Transcription Factors In A Neuronal Gene Regulatory Network., Gregory M Miller, Babatunde A Ogunnaike, James S Schwaber, Rajanikanth Vadigepalli Jan 2010

Robust Dynamic Balance Of Ap-1 Transcription Factors In A Neuronal Gene Regulatory Network., Gregory M Miller, Babatunde A Ogunnaike, James S Schwaber, Rajanikanth Vadigepalli

Department of Pathology, Anatomy, and Cell Biology Faculty Papers

BACKGROUND: The octapeptide Angiotensin II is a key hormone that acts via its receptor AT1R in the brainstem to modulate the blood pressure control circuits and thus plays a central role in the cardiac and respiratory homeostasis. This modulation occurs via activation of a complex network of signaling proteins and transcription factors, leading to changes in levels of key genes and proteins. AT1R initiated activity in the nucleus tractus solitarius (NTS), which regulates blood pressure, has been the subject of extensive molecular analysis. But the adaptive network interactions in the NTS response to AT1R, plausibly related to the development of …