Life Sciences Commons^™

Understanding The Intracellular Positive Regulatory Interactions In The Metabolic Network, Sultana Mohammed Al Zubaidi

Theses

Metabolism is an essential cellular process that produces essential components such as energy molecules and cellular building blocks. It is regulated by intracellular metabolite-enzyme interactions, which can increase or decrease enzyme efficiency. Each metabolic process is not evolved in nature and may not contribute to the optimal efficiency of the organism's metabolism. However, this study aims to clarify the relationship between cellular positive regulatory interactions and the cell's metabolic network, especially how these relationships regulate metabolism and how it evolved in nature. To obtain that, for each enzyme, the activation molecules from the BRENDA database were downloaded, mapped data to …

Exploring Bioprocessing Technologies For Diverse Industrial Application Of Canola, Ahmad Fawzi N Alhomodi

Electronic Theses and Dissertations

Globally, canola is the second largest oilseed crop after soybean, which is processed for the commercial production of high value oil. The industrial processing steps for canola oil extraction include preprocessing (cleaning, pressing, flaking, and cooking), mechanical pressing, and/or organic solvents. This process results in large quantities of protein-rich (~ 40% dry basis) meal as a co-product. The meal is used partially in animal diet (<30% inclusion) due to high levels of antinutritional factors (ANFs) such as high fibers, phytic acid, and glucosinolates, and low metabolizable energy. Thus, this research was designed to examine various bioprocessing technologies (i.e., traditional sprouting, solid state fermentation, submerged state fermentation, co-culture fermentation, mild pretreatments, and their combinations) for possible advancement in canola use. Canola seed sprouting for 6-day period led to an increase in protein content and a reduction in ANFs and oil content of sprouts compared to ungerminated seed. Subsequent submerged state fermentation of 6- day old sprouts using three different strains (Aureobasidium pullulans, Trichoderma reesei and Neurospora crassa) further increased protein content and lowered ANFs. Solid-state fermentation of 144 h old sprouts using A. pullulans, N. crassa and T. reesei enhanced the protein content and reduced ANFs of sprouts. Sprouting canola seed for three days helped in hull removal, leading to high protein meal accompanied by low fiber and phytic acid level. Three-day seed sprouting had no effect on oil yield, but free fatty acid content was higher compared to seed oil. Co-culture fermentation of HECM under solid state process showed maximum reduction in fiber content with co-culture of A. pullulans and N. crassa while a combination of A. pullulans and T. reesei promoted the highest GLS and phytic acid reductions compared to other combinations, which indicated the advantage of coculture inoculation over monoculture in terms of ANFs reduction. Mild pretreatment of HECM using deionized water resulted in washed HECM (WHECM) with lower soluble sugars and GLS compared to untreated HECM, whereas protein and amino acid were concentrated due to the removal of soluble components. WHECM compared to HECM showed higher protein digestibility when fed to rainbow trout. Subsequent mono- and coculture fermentation of HECM and WHECM under submerged process resulted in higher protein and amino acid content and lower ANFs levels compared to uninoculated controls. The results of cellulase, endoglucanase and β-glucosidase activity indicated the crucial role of used substrates, fungi, fermentation modes (solid state/submerged stated) and inoculation methods (mono-/co-culture) on enzyme activities.

The Effect Of Alcalase Concentration On The Proteins From The Shells Of Litopenaeus Setiferus (White Shrimp), Liam T. Quan

Journal of the South Carolina Academy of Science

Chitin is a naturally abundant polymer that also happens to be biodegradable. Chitin can be used in a variety of different products such as biodegradable plastics, papers, medical products, foods, and medical treatments. To extract chitin, shells must be demineralized and deproteinized. The goal of this experiment was to examine the effect of the protease Alcalase in the deproteinization of litopenaeus setiferus shells. The hypothesis was that if the concentration of Alcalase increased, then the absorbance of proteins in the spectrophotometer reading would increase. The null hypothesis was that if the concentration increased there would be no change in absorption. …

Lipase From New Isolate Bacillus Cereus Ata179: Optimization Of Production Conditions, Partial Purification, Characterization And Its Potential In The Detergent Industry, Eli̇f Demi̇rkan, Aynur Aybey Çeti̇nkaya, Maoulida Abdou

Turkish Journal of Biology

In this study, 341 Bacillus sp. strains were isolated from agricultural soils of Turkey. The potent extracellular lipase producer was selected. It was identified by 16S rRNA, named as Bacillus cereus ATA179. Optimal nutritional and physical parameters for lipase production were determined. Sucrose as the carbon source, (NH4)2HPO4 as the nitrogen source, CaCl2 as the metal ion were obtained. The best results of physical parameters were stated at 45°C, pH 7.0, shaking rate 50 rpm, inoculation amount 7% and inoculum age 24 h. ATA179 strain showed a 51% increase in enzyme production in the modified medium created by optimizing nutritional …

Soil Chemistry And Enzyme Data Examining Impacts Of Callery Pear Stem Cutting, Michaela J. Woods, Grace K. Attea, Ryan W. Mcewan

Five Rivers MetroParks Collaboration Data Archive

This dataset encompasses information following the collection of soil from underneath cut and treated Callery pear stems and untreated Callery pear trees. Soil was assessed for moisture, pH, and enzyme activities.

Comparison Of An Electron Transport System (Ets) Enzyme-Mediated Reduction Assay And Respiration Rate Of The Invasive Copepod Eurytemora Carolleeae In Green Bay, Wi, Usa, Alexander Timpe

Lawrence University Honors Projects

The use of aquatic resources for agriculture, trade, and recreation adds stress to water-dwelling organisms. Rapid changes in abiotic conditions, such as warming due to climate change and nutrient loading from agricultural runoff and urban areas, threaten to induce profound alterations to aquatic environments. These changes affect interspecific community interactions and may cause an aquatic resource to lose its functionality that is valuable to humans. Studying organisms such as plankton that form an ecosystem’s foundation is an important step towards understanding the entire food web and predicting how it may or may not be able to respond to a changing …

Theobromine And Related Methylxanthines As Inhibitors Of Primary Amine Oxidase, Padraig Shanahan, Jeffrey O’Sullivan, Keith Tipton, Gemma Kinsella, Barry Ryan, Gary Henehan

Articles

Methylxanthines are the most widely consumed drugs in the world and evidence of their health benefits has been growing in recent years. Primary Amine Oxidase (PrAO) has been recognised as a therapeutic target for amelioration of inflammatory, vascular and neurodegenerative diseases. Previous work in our laboratories showed that caffeine inhibited Bovine PrAO with a Ki of 1.0mM using benzylamine as substrate.

This study aimed to extend our previous work and explore the possibility that related methylxanthines might influence PrAO activity. While paraxanthine, theophylline and 7-methylxanthine had little effect on PrAO, theobromine was a noncompetitive inhibitor with a Ki of 276±44µM. …

Lonicera Maackii Alters Decay Dynamics Of Coarse Woody Debris, Michaela J. Woods

Browse all Theses and Dissertations

Since industrialization, anthropogenic carbon emissions have led to excess atmospheric carbon dioxide that may alter the stability of ecosystem processes. Microorganisms are essential in mitigating excess carbon and play a notable role in the breakdown of organic material. This process, decomposition, is essential in forested ecosystems where microorganisms can recycle nutrients and store carbon in soil organic matter or release it through respiration. Fungi participate in decomposition through the release of enzymes responsible for carrying out the chemical reactions that break down plant material. Species introductions have the potential to alter decomposition dynamics. In the Midwestern US, the invasive shrub …

A Study On The Interactions Of Trehalose With Model Folate Compounds, Carolyn R Ware

Chancellor’s Honors Program Projects

No abstract provided.

The Urea Carboxylase And Allophanate Hydrolase Activities Of Urea Amidolyase Are Functionally Independent, Yi Lin, Cody J. Boese, Martin St. Maurice

Biological Sciences Faculty Research and Publications

Urea amidolyase (UAL) is a multifunctional biotin-dependent enzyme that contributes to both bacterial and fungal pathogenicity by catalyzing the ATP-dependent cleavage of urea into ammonia and CO₂. UAL is comprised of two enzymatic components: urea carboxylase (UC) and allophanate hydrolase (AH). These enzyme activities are encoded on separate but proximally related genes in prokaryotes while, in most fungi, they are encoded by a single gene that produces a fusion enzyme on a single polypeptide chain. It is unclear whether the UC and AH activities are connected through substrate channeling or other forms of direct communication. Here, we use …

The Role Of Biotin And Oxamate In The Carboxyltransferase Reaction Of Pyruvate Carboxylase, Adam D. Lietzan, Yi Lin, Martin St. Maurice

Biological Sciences Faculty Research and Publications

Pyruvate carboxylase (PC) is a biotin-dependent enzyme that catalyzes the MgATP-dependent carboxylation of pyruvate to oxaloacetate, an important anaplerotic reaction in central metabolism. During catalysis, carboxybiotin is translocated to the carboxyltransferase domain where the carboxyl group is transferred to the acceptor substrate, pyruvate. Many studies on the carboxyltransferase domain of PC have demonstrated an enhanced oxaloacetate decarboxylation activity in the presence of oxamate and it has been shown that oxamate accepts a carboxyl group from carboxybiotin during oxaloacetate decarboxylation. The X-ray crystal structure of the carboxyltransferase domain from Rhizobium etli PC reveals that oxamate is positioned in the active site …

Functionally Diverse Biotin-Dependent Enzymes With Oxaloacetate Decarboxylase Activity, Adam D. Lietzan, Martin St. Maurice

Biological Sciences Faculty Research and Publications

Biotin-dependent enzymes catalyze carboxylation, decarboxylation and transcarboxylation reactions that participate in the primary metabolism of a wide range of organisms. In all cases, the overall reaction proceeds via two half reactions that take place in physically distinct active sites. In the first half-reaction, a carboxyl group is transferred to the 1-N′ of a covalently tethered biotin cofactor. The tethered carboxybiotin intermediate subsequently translocates to a second active site where the carboxyl group is either transferred to an acceptor substrate or, in some bacteria and archaea, is decarboxylated to biotin and CO₂ in order to power the export of sodium …

An Investigation Into The Relationship Between The Activation Of Amp Kinase And The Acetylation Of Microtubules, Jessica Sarrantonio

Honors Theses

Microtubules, or cytoskeletal polymers composed of the protein tubulin, form long hollow tubes in the cell and are responsible for many critical roles. Previous research has shown that depletion of ATP causes microtubules to become stable, i.e. resistant to depolymerization. It has also been shown that enhanced stability of microtubules correlates with increased tubulin acetylation, a common microtubule posttranslation modification. ATP-depletion is a severe metabolic stressor, and as such, we expect that this treatment would activate AMP kinase, an enzyme deemed the “master regulator” of metabolism. When activated by a variety of stressors, this enzyme can initiate a program of …

Tissue-Specific Thyroid Hormone Regulation Of Gene Transcripts Encoding Iodothyronine Deiodinases And Thyroid Hormone Receptors In Striped Parrotfish (Scarus Iseri), Kaitlin M. Johnson, Sean C. Lema

Biological Sciences

In fish as in other vertebrates, the diverse functions of thyroid hormones are mediated at the peripheral tissue level through iodothyronine deiodinase (dio) enzymes and thyroid hormone receptor (tr) proteins. In this study, we examined thyroid hormone regulation of mRNAs encoding the three deiodinases dio1, dio2 and dio3 – as well as three thyroid hormone receptors trαA, trαB and trβ – in initial phase striped parrotfish (Scarus iseri). Parrotfish were treated with dissolved phase T₃ (20 nM) or methimazole (3 mM) for 3 days. Treatment with exogenous T₃ elevated circulating T₃, while the …

Characterization Of The Substrate Specificity And Mechanism Of Protein Arginine Methyltransferase 1, Whitney Lyn Wooderchak

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protein arginine methyltransferases (PRMTs) posttranslationally modify protein arginine residues. Type I PRMTs catalyze the formation of monomethylarginine (MMA) and asymmetric dimethylarginine (ADMA) via methyl group transfer from S-adenosyl methionine onto protein arginine residues. Type II PRMTs generate MMA and symmetric dimethylarginine. PRMT-methylation affects many biological processes. Although PRMTs are vital to normal development and function, PRMT-methylation is also linked to cardiovascular disease, stroke, multiple sclerosis, and cancer.

Thus far, nine human PRMT isoforms have been identified with orthologues present in yeast, plants, and fish. PRMT1 predominates, performing an estimated 85% of all protein arginine methylation in vivo. Yet, the substrate …

Exogenous Expression Of Human Na, K-Atpase Isoforms In Insect Cells, Christal Renee' Carpenter

McCabe Thesis Collection

The Na-K-ATPase or Na pump is a membrane bound enzyme that maintains a high concentration of K+ and low concentration of Na+ in most animal cells. Three subunits constitute the Na pump, named alpha (ex), beta (~). and gamma (y). The ex subunit is responsible for the catalytic and transport of the enzyme, the ~ subunit is important for bringing the ex to the plasma membrane of the cell, and y is a regulatory subunit of the Na pump. In addition, there are several molecular variants or isoforms of the ex and ~ subunits. At present four different ex (ex …

Investigation On The Liver Functions Agriculturel Workers Chronically Exposed To Pesticides, Ülkü Çömelekoğlu, Bi̇rgül Mazmanci, Abdullah Arpaci

Turkish Journal of Biology

Blood samples were taken from agricultural workers who had been exposed to pesticides for long periods in Içel and the surrounding agricultural areas (n=150) and levels of AST, ALT, ALP, LDH, total protein, albumin and globulin in serum were determined. The same parameters were measured in 64 persons who are not exposed to pesticides in any way. Study group showed significantly higher levels of AST, ALT, ALP and LDH (p

Esterase D And Abo Polymorphisms In Turkish Population, E. Ferda Perçin, Ilhan Sezgin, Ahmet Çolak, Ziynet Çinar

Turkish Journal of Biology

This study is intended to be a contribution to the determination of the genetic structure of the Turkish population. For this purpose, esterase D (ESD), an erythrocyte enzyme, and ABO blood group loci phenotypes from the blood samples of two hundred healthy and unrelated individuals from different regions of Turkey were determined. ESD enzyme phenotypes were determined by cellulose acetate electrophoresis, and ABO blood group phenotypes were determined by the aglutination test; in this way, allele frequencies were calculated. It was determined that the allele frequencies of the ESD enzyme were ESD*1=0.807±0.019, and ESD*2=0.193±0.019. The allele frequencies of the ABO …

Development Of An Enzyme Linked Immunosorbent Assay Capable Of Detecting Antibodies Specific To The Cytomegalovirus, Rachel Bird

Undergraduate Honors Capstone Projects

The cytomegalovirus (CMV) is capable of causing serious illness and death in immunocompromised individuals. The objective of this research is to develop a method of detecting antibodies against CMV. This has lead to the development of an Enzyme linked Immunosorbent Assay (ELISA) that is capable of detecting CMV or anti-CMV antibodies. The process in the development of such an assay involves; 1) the purification of Hyperimmune Rabbit Serum containing anti-CMV antibodies, 2) determination of the appropriate concentrations necessary to provide a clear and concise response to the test, and 3) testing of monoclonal antibodies to determine their specificity towards the …

The Primary Structure Of A Fungal Chitin Deacetylase Reveals The Function For Two Bacterial Gene Products., Dimitris Kafetzopoulos, George Thireos, John N. Vournakis, Vassilis Bouriotis

Dartmouth Scholarship

Chitin deacetylase (EC 3.5.1.41) hydrolyzes the N-acetamido groups of N-acetyl-D-glucosamine residues in chitin. A cDNA to the Mucor rouxii mRNA encoding chitin deacetylase was isolated, characterized, and sequenced. Protein sequence comparisons revealed significant similarities of the fungal chitin deacetylase to rhizobial nodB proteins and to an uncharacterized protein encoded by a Bacillus stearothermophilus open reading frame. These data suggest the functional homology of these evolutionarily distant proteins. NodB is a chitooligosaccharide deacetylase essential for the biosynthesis of the bacterial nodulation signals, termed Nod factors. The observed similarity of chitin deacetylase to the B. stearothermophilus gene product suggests that this gene …

Functional And Morphological Studies Of Mitochondria Exposed To Undecagold Clusters: Biologic Surfaces Labeling With Gold Clusters, E. Valdivia, C. Gabel, J. E. Reardon, C. A. Cajacob, H. Yang, R. S. Wehbi, G. L. Scott, P. A. Frey, L. A. Fahien

Scanning Microscopy

This study reports morphological and functional alterations observed in respiring isolated mitochondria when they are exposed to nonpenetrating, positive electrostatically charged synthetic undecagold clusters. Modification of the undecagold clusters positive charges change or prevent the functional effects and the binding to the outside surface of the mitochondria. The mitochondrial functional alterations are dependent on the oxidative phosphorylation capacity of the isolated organelles. The results of these experiments indicate that artificial undecagold may be useful to explore the molecular mechanisms of biological energy transducers which require electric charges separation, ionic fluxes, and electric surface properties.

Incorporation Of Protoporphyrin Ix Into Chlorophyll A And Purification And Properties Of (-) S-Adenosyl-L-Methionine-Magnesium Protoporphyrin Ix Methyltransferase, Jii Shieh

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Experiments on the incorporation of ¹⁴C-protoporphyrin IX into chlorophyll indicate that ferrous iron may be involved in reactions between protoporphyrin IX and chlorophyll. Experiments using both health and chlorotic tobacco tissue for incorporation studies show that incorporation of protoporphyrin IX into chlorophyll was activated by Fe²⁺.

S-adenosylmethionine-magnesium protoporphyrin IX methyltransferase (E.C.2.1.1.11) which catalyzes the reaction between S-adenosylmethionine and Mg protoporphyrin IX to form S-adenosylhomocysteine and Mg protoporphyrin IX monomethyl ester was demonstrated in barley seedlings. Methyltransferase was found principally in whole chloroplasts and fractions containing mitochondria were disintegrated chloroplasts. Use of detergent Tween 80, precipitation with protamine …

Ribose 5'-Phosphate Aminotransferase Of Salmonella Typhimurium, Gulamnabi Y. Vahora

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Ribose-5 '-phosphate aminotransferase, an alternate first enzyme in the purine de novo pathway, has been purified about 800-fold from Salmonella typhimurium (pur D-55 ). The enzyme is distinct from ribosylpyrophosphate-5'-phosphate amidotransferase (E C 2.4.2.14), has an approximate molecular weight of 229,000, and requires ribose-5'-phosphate, ATP and either ammonium ion, carbamyl phosphate or L-glutamine, to synthesize 5'-phosphoribosylamine (PRA). A coupled assay system, employing a S. typhimurium (pur G-310) extract as the coupling-agent source, was used to measure the enzyme. PRA production in this system was inhibited by higher ribose-5' -phosphate concentrations and longer (over 10 minutes) incubation periods. It …