Biotechnology Commons^™

The Use Of Co-Culturing In Solid Substrate Cultivation And Possible Solutions To Scientific Challenges, Wanying Yao, Sue E. Nokes

Biosystems and Agricultural Engineering Faculty Publications

This perspective systematically summarizes the use of solid substrate co‐cultures in agriculture, food, plant, and industrial biotechnology applications. The summarization is organized by organism, i.e. fungus, bacteria, yeast and then co‐cultivation of either two or three organisms. Generally, in solid substrate co‐culture, the organisms synergistically penetrate and degrade the solid substrate, thereby increasing product yield and productivity over a monoculture. Efforts to increase co‐culture performance include optimizing process parameters (pH, temperature, moisture, and oxygen demand) and defining the acceptable types of substrate. Scientific challenges exist in understanding the interactions between microbial stains, such as viability, suite of products, and bio‐transformations. …

A Morphological Analysis Of A Hybrid Swarm Of Native Ulmus Rubra And Introduced U. Pumila (Ulmaceae) In Southeastern Nebraska, Christian Elowsky, Ingrid E. Jordon-Thaden, Robert B. Kaul

Nebraska Center for Biotechnology: Faculty and Staff Publications

The parental species and hybrid swarm of native Ulmus rubra Muhl. and the introduced, naturalized and weedy U. pumila L. were investigated in a 65-km transect in Lancaster, Saunders, and Butler counties in Nebraska. Thirty-two trees of U. rubra, 32 of U. pumila, and 50 of the hybrid swarm were sampled for leaves and buds and subsampled for flowers and fruits. Leaves were measured for petiole length, blade length, width, primary and secondary teeth per cm, number of secondary teeth per primary tooth, and texture. Buds were scored for color and distribution of trichomes. Flowers were sampled for stamen counts …

Influence Of Media Composition On The Growth Rate Of Chlorella Vulgaris And Scenedesmus Acutus Utilized For Co2 Mitigation, Czarena L. Crofcheck, Xinyi E, Aubrey Shea, Michael D. Montross, Mark Crocker, Rodney Andrews

Biosystems and Agricultural Engineering Faculty Publications

Atmospheric carbon dioxide levels have increased since the industrial revolution due to increasing combustion of fossil fuels. One possible CO₂ mitigation strategy is the use of microalgae for mitigation of CO₂. This paper focuses on the influence of media composition on the growth rate of two microalgae strains, Chlorella vulgaris and Scenedesmus actus. A KNO₃ based medium was found to work better for Chlorella, while a urea based equivalent worked better for Scenedesmus. The urea based media investigated here resulted in growth similar to that found with previously reported KNO₃ based media. …

Overview Of The Recent Fda Process Validation Guidance For Medicinal Product Development And Manufacture, Nuala Calnan (Editor)

Conference Papers

This conference paper presents an overview of the recently published FDA Process Validation Guideline for the development and manufacture of medicinal drug products. It discusses the impacts of conducting life cycle based validation activities and highlights the future challenges in meeting the ongoing 'Continued Process Validation' (CPV) expectations of the regulator. It also presents the latest information on the EU draft Process Validation Guidance - due for publication in late 2013 / 2014.

Use Of Image Cytometry For Quantification Of Pathogenic Fungi In Association With Host Cells, Charlotte A. Berkes, Leo Li-Ying Chan, Alisha Wilkinson, Benjamin Paradis

Biology Faculty Publications

Studies of the cellular pathogenesis mechanisms of pathogenic yeasts such as Candida albicans, Histoplasma capsulatum, and Cryptococcus neoformans commonly employ infection of mammalian hosts or host cells (i.e. macrophages) followed by yeast quantification using colony forming unit analysis or flow cytometry. While colony forming unit enumeration has been the most commonly used method in the field, this technique has disadvantages and limitations, including slow growth of some fungal species on solid media and low and/or variable plating efficiencies, which is of particular concern when comparing growth of wild-type and mutant strains. Flow cytometry can provide rapid quantitative information regarding yeast …

Diversity Of Plant Methionine Sulfoxide Reductases B And Evolution Of A Form Specific For Free Methionine Sulfoxide, Dung Tien Le, Lionel Tarrago, Yasuko Watanabe, Alaattin Kaya, Byung Cheon Lee, Uyen Tran, Rie Nishiyama, Dmitri E. Fomenko, Vadim N. Gladyshev, Lam-Son Phan Tran

Department of Biochemistry: Faculty Publications

Methionine can be reversibly oxidized to methionine sulfoxide (MetO) under physiological conditions. Organisms evolved two distinct methionine sulfoxide reductase families (MSRA & MSRB) to repair oxidized methionine residues. We found that 5 MSRB genes exist in the soybean genome, including GmMSRB1 and two segmentally duplicated gene pairs (GmMSRB2 and GmMSRB5, GmMSRB3 and GmMSRB4). GmMSRB2 and GmMSRB4 proteins showed MSRB activity toward protein-based MetO with either DTT or thioredoxin (TRX) as reductants, whereas GmMSRB1 was active only with DTT. GmMSRB2 had a typical MSRB mechanism with Cys121 and Cys 68 as catalytic and resolving residues, respectively. Surprisingly, this enzyme also possessed …

Dna Purification Process Optimization At Life Technologies Corporation, Trevor J. Shepherd

Master's Theses

This project focused on optimizing the plasmid DNA purification process at Life Technologies. These plasmids are designed to code for specialized proteins used by research universities, national laboratories, or research companies. Once cultivated and harvested, the plasmids must be analyzed for quality and quantity. The project is divided into improving three aspects of the process: 1) plasmid identification, 2) plasmid purity evaluation, and 3) process yield. Plasmid identification is now simpler, more robust and has zero ambiguity. Plasmid purity evaluation is now measured with computer software, which reduces user error and eliminates subjectivity. Using the nascent metrics provided by the …

Validation Of Antibodies Used To Study Hypoxia Inducible Factors In Two Species Of Fundulus, Jenna D. Hill

LSU New Orleans Theses and Dissertations

Hypoxia inducible factors (HIFs) are transcription factors and the master regulators of oxygen-dependent gene expression in animals. The focus of this thesis is the distribution of HIF protein in tissues of the fish Fundulus heteroclitus and F. grandis, two widespread species that occur in naturally hypoxic waters. Polyclonal antibodies against HIF-1α, HIF-2α, and HIF-3α were tested on proteins made in vitro and on extracts made from several tissues of normoxic and hypoxic fish. Antibodies against HIF-1α and 3α bound specifically to full length protein made in vitro, and produced bands on western blots of nuclear extracts of near …

Regulation Of The Target Of Rapamycin Signaling Pathway In Saccharomyces Cerevisiae, Tammy Pracheil

LSU New Orleans Theses and Dissertations

An integrative, biochemical, genetic, and molecular biology approach utilizing gene manipulation, gene knock outs, plasmid based protein expression, and in vivo protein localization of fluorescence tagged proteins was employed to determine the function of an essential protein, Lst8, in TORC1 and TORC2 signaling and a previously uncharacterized complex, the Far3-7-8-9-10-11 complex (Far complex) in the budding yeast, Saccharomyces cerevisiae. Mutations in SAC7 and FAR11 suppressed lethality of both lst8 and tor2-21 mutations but not TORC1 inactivation, suggesting that the essential function of Lst8 is linked only to TORC2.

Far11, a component of a six-member complex, was found to interact …

Aerobic Degradation Of Α-, Β-, Γ-Hexachlorocyclohexane By Narragansett Bay Bacterioplankton, Ian M. Rambo

Senior Honors Projects

Hexachlorocyclohexanes (HCHs) are a family of chlorinated organic compounds that were previously used as agricultural insecticides. HCHs are recognized as persistent organic pollutants due to their toxicity, recalcitrant properties, and tendency to bioaccumulate in food webs. Although HCH was first synthesized in 1825, its use was not widespread until the discovery of the insecticidal activity of the γ-HCH isomer in 1942. γ-HCH and its toxic waste isomers α-HCH and β-HCH were banned from production and use by the United Nations in 2009, yet these chemicals still present environmental problems due to their persistence in soils and surface waters. HCHs continue …

Biomass Gasification: An Alternative Solution To Animal Waste Management, Hanjing Wu

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

The overall goal of this research was to evaluate gasification of animal waste as an alternative manure management strategy, from the standpoints of syngas production and biochar application.

To meet the overall objective, the thermogravimetric characteristics of dairy manure, as a thermochemical conversion feedstock, were studied firstly. Then, gasification technology was applied to dairy manure and feedlot manure using a fluidized-bed laboratory-scale gasifier. In addition, biochar derived from the feedlot manure was examined for its effects on nutrient leaching as a soil amendment. Finally, a life cycle assessment was conducted to evaluate greenhouse gas emissions of two feedlot manure management …

A Cell Simulator Platform: The Cell Collective, Tomáš Helikar, B. Kowal, J. A. Rogers

Department of Biochemistry: Faculty Publications

Diseases are often a result of multiple malfunctions in complex, nonlinear biological/biochemical networks. As such, these processes are far more complicated to understand because they tend to give rise to functions that are emergent in nature, i.e., higher-level (mal)functions that are more than the sum of their parts. Systems biology provides a new approach to understanding biological systems and diseases from a holistic perspective.

Expression, Production, And Purification Of Novel Therapeutic Proteins, Mckinzie Shea Fruchtl

Graduate Theses and Dissertations

Interest in the production of recombinant proteins consisting of collagen binding domain (CBD) fused to a bioactive material has increased due to the targeting/attachment capabilities of CBD. For example, CBD fusions can be applied to the reversing of bone density loss and the repair of the eardrum, specifically, by choosing an appropriate fusion partner (parathyroid hormone or epidermal growth factor). The production of CBD fusions was examined using batch and fed-batch culturing of Escherichia coli to express the fusion proteins, and affinity chromatography to isolate the final product.

Different medium formulations, feeding strategies, and induction methods were tested in order …

The Geometry And Sensitivity Of Ion-Beam Sculpted Nanopores For Single Molecule Dna Analysis, Ryan Connor Rollings

Graduate Theses and Dissertations

In this dissertation, the relationship between the geometry of ion-beam sculpted solid-state nanopores and their ability to analyze single DNA molecules using resistive pulse sensing is investigated. To accomplish this, the three dimensional shape of the nanopore is determined using energy filtered and tomographic transmission electron microscopy. It is shown that this information enables the prediction of the ionic current passing through a voltage biased nanopore and improves the prediction of the magnitude of current drop signals when the nanopore interacts with single DNA molecules. The dimensional stability of nanopores in solution is monitored using this information and is improved …

Steered Molecular Dynamics Simulations Of A Type Iv Pilus Probe Initial Stages Of A Force-Induced Conformational Transition, Joseph L. Baker, Nicolas Biais, Florence Tama

Publications and Research

Type IV pili are long, protein filaments built from a repeating subunit that protrudes from the surface of a wide variety of infectious bacteria. They are implicated in a vast array of functions, ranging from bacterial motility to microcolony formation to infection. One of the most well-studied type IV filaments is the gonococcal type IV pilus (GC-T4P) from Neisseria gonorrhoeae, the causative agent of gonorrhea. Cryo-electron microscopy has been used to construct a model of this filament, offering insights into the structure of type IV pili. In addition, experiments have demonstrated that GC-T4P can withstand very large tension forces, and …

An Integrated Research Approach To The Development Of Fungal Isolates As Bioherbicides For The Control Of Woody Weeds, Patrick Mershon

Independent Study Project (ISP) Collection

Prickly acacia (Acacia nilotica) is an invasive weed tree in Australia, ranked seventh on the list of Weeds of National Significance because of its aggressiveness and its economic and environmental impacts. The significant downsides to manual and chemical control of this species drove the search for a biological control mechanism. Surveying and sampling stands of A. nilotica naturally exhibiting disease symptoms yielded several native Australian fungi capable of pathogenesis in this species, which are currently under assessment as potential bioherbicides for this weed. Three field trials were established at the same site in order to further advance current …

The Unstable Ground Of Low Hierarchies, Joshua Dinsmore

The Transdisciplinary STEAM+ Journal

Broad Vision is a collaborative project between the Sciences and Arts. It involves students and lecturers from six different departments, across three schools at the University of Westminster, London, UK. In the first year of the project we worked with the microscope as the locus for our interconnections.

A Single Point Mutation In The Listerial Betl Sigma(A)-Dependent Promoter Leads To Improved Osmo- And Chill-Tolerance And A Morphological Shift At Elevated Osmolarity, Ronald F. Hoffman, Susan Mclernon, Audrey Feeney, Colin Hill, Roy D. Sleator

Department of Biological Sciences Publications

Betaine uptake in Listeria monocytogenes is mediated by three independent transport systems, the simplest of which in genetic terms is the secondary transporter BetL. Using a random mutagenesis approach, based on the E. coli XL1 Red mutator strain, we identified a single point mutation in a putative promoter region upstream of the BetL coding region which leads to a significant increase in betL transcript levels under osmo- and chill-stress conditions and a concomitant increase in stress tolerance. Furthermore, the mutation appears to counter the heretofore unreported “twisted” cell morphology observed for L. monocytogenes grown at elevated osmolarities in tryptone soy …

Detection And Quantification Of Methylation In Dna Using Solid-State Nanopores., Jiwook Shim, Gwendolyn I Humphreys, Bala Murali Venkatesan, Jan Marie Munz, Xueqing Zou, Chaitanya Sathe, Klaus Schulten, Farhad Kosari, Ann M Nardulli, George Vasmatzis, Rashid Bashir

College of Science & Mathematics Departmental Research

Epigenetic modifications in eukaryotic genomes occur primarily in the form of 5-methylcytosine (5 mC). These modifications are heavily involved in transcriptional repression, gene regulation, development and the progression of diseases including cancer. We report a new single-molecule assay for the detection of DNA methylation using solid-state nanopores. Methylation is detected by selectively labeling methylation sites with MBD1 (MBD-1x) proteins, the complex inducing a 3 fold increase in ionic blockage current relative to unmethylated DNA. Furthermore, the discrimination of methylated and unmethylated DNA is demonstrated in the presence of only a single bound protein, thereby giving a resolution of a single …

Functional Heterologous Expression Of An Engineered Full Length Cipa From Clostridium Thermocellum In Thermoanaerobacterium Saccharolyticum, Devin H. Currie, Christopher D. Herring, Adam M. Guss, Daniel G. Olson, David A. Hogsett, Lee R. Rynd

Dartmouth Scholarship

Background: Cellulose is highly recalcitrant and thus requires a specialized suite of enzymes to solubilize it into fermentable sugars. In C. thermocellum, these extracellular enzymes are present as a highly active multi-component system known as the cellulosome. This study explores the expression of a critical C. thermocellum cellulosomal component in T. saccharolyticum as a step toward creating a thermophilic bacterium capable of consolidated bioprocessing by employing heterologously expressed cellulosomes. Results:We developed an inducible promoter system based on the native T. saccharolyticum xynA promoter, which was shown to be induced by xylan and xylose. The promoter was used to …