{"id":{"repo_id":"lethbridge","oai_identifier":"oai:opus.uleth.ca:10133/4763"},"canonical_url":"https://search.dev.ndltd.org/etd/lethbridge/oai:opus.uleth.ca:10133/4763","repository":{"repo_id":"lethbridge","name":"University of Lethbridge","base_url":"https://opus.uleth.ca/server/oai/request"},"display":{"title":"Rapid kinetic studies of PhyA from Selenomonas ruminantium, and a simplified means of production of a phosphate biosensor","abstract":"Myo-inositol polyphosphates (IPs) are ubiquitous in nature and involved in various cellular events. Dephosphorylation of IPs by protein tyrosine phosphatase-like polyphosphatases (PTPLPs) occurs via a complex pathway, and the in vivo function of many of these enzymes remains unknown. In order to further our understanding of PTPLP catalyzed dephosphorylation of IPs; I present rapid kinetics studies of the representative PTPLP PhyA from Selenomonas ruminantium (PhyAsr). These studies revealed kinetic parameters of PhyAsr dimerization, and myo-inositol hexakisphosphate (IP6) binding to the homodimer. In addition to studying PhyAsr, I have developed a simplified methodology to produce a biosensor capable of detecting phosphate release in real-time. The phosphate biosensor is fluorescently labeled and utilizes Escherichia coli Phosphate-binding protein (PhoS). The results show that my proposed methodology yields a functional biosensor and is feasible for large-scale production. I envision this methodology to be versatile and useful for a large number of research applications where detection of free phosphate is required.","abstract_html":"Myo-inositol polyphosphates (IPs) are ubiquitous in nature and involved in various cellular events. Dephosphorylation of IPs by protein tyrosine phosphatase-like polyphosphatases (PTPLPs) occurs via a complex pathway, and the in vivo function of many of these enzymes remains unknown. In order to further our understanding of PTPLP catalyzed dephosphorylation of IPs; I present rapid kinetics studies of the representative PTPLP PhyA from Selenomonas ruminantium (PhyAsr). These studies revealed kinetic parameters of PhyAsr dimerization, and myo-inositol hexakisphosphate (IP6) binding to the homodimer. In addition to studying PhyAsr, I have developed a simplified methodology to produce a biosensor capable of detecting phosphate release in real-time. The phosphate biosensor is fluorescently labeled and utilizes Escherichia coli Phosphate-binding protein (PhoS). The results show that my proposed methodology yields a functional biosensor and is feasible for large-scale production. I envision this methodology to be versatile and useful for a large number of research applications where detection of free phosphate is required.","abstract_has_math":false,"creators":["Smith, Dustin D.","University of Lethbridge. Faculty of Arts and Science"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016","date_published":"2016","updated_at":"2026-07-27T20:02:35Z","subjects":["Biochemistry","Enzyme kinetics","Phosphate sensor","PTPLP","myo-inositol","Phosphatase","Stopped-flow","PhyA","PhoS","Phosphate release","Phosphate binding protein","Phytase"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["hdl:10133/4763"],"render_values":[{"text":"hdl:10133/4763","href":null,"code":true}]}]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.issued","label":"Date","values":["2016"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biochemistry","Enzyme kinetics","Phosphate sensor","PTPLP","myo-inositol","Phosphatase","Stopped-flow","PhyA","PhoS","Phosphate release","Phosphate binding protein","Phytase"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["hdl:10133/4763"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.other","label":"Dc Description Other","values":["Myo-inositol polyphosphates (IPs) are ubiquitous in nature and involved in various cellular events. Dephosphorylation of IPs by protein tyrosine phosphatase-like polyphosphatases (PTPLPs) occurs via a complex pathway, and the in vivo function of many of these enzymes remains unknown. In order to further our understanding of PTPLP catalyzed dephosphorylation of IPs; I present rapid kinetics studies of the representative PTPLP PhyA from Selenomonas ruminantium (PhyAsr). These studies revealed kinetic parameters of PhyAsr dimerization, and myo-inositol hexakisphosphate (IP6) binding to the homodimer. In addition to studying PhyAsr, I have developed a simplified methodology to produce a biosensor capable of detecting phosphate release in real-time. The phosphate biosensor is fluorescently labeled and utilizes Escherichia coli Phosphate-binding protein (PhoS). The results show that my proposed methodology yields a functional biosensor and is feasible for large-scale production. I envision this methodology to be versatile and useful for a large number of research applications where detection of free phosphate is required."]},{"key":"dc:title","label":"Title","values":["Rapid kinetic studies of PhyA from Selenomonas ruminantium, and a simplified means of production of a phosphate biosensor"]}]}],"canonical_facts":{"dc:date.issued":["2016"],"dc:description.other":["Myo-inositol polyphosphates (IPs) are ubiquitous in nature and involved in various cellular events. Dephosphorylation of IPs by protein tyrosine phosphatase-like polyphosphatases (PTPLPs) occurs via a complex pathway, and the in vivo function of many of these enzymes remains unknown. In order to further our understanding of PTPLP catalyzed dephosphorylation of IPs; I present rapid kinetics studies of the representative PTPLP PhyA from Selenomonas ruminantium (PhyAsr). These studies revealed kinetic parameters of PhyAsr dimerization, and myo-inositol hexakisphosphate (IP6) binding to the homodimer. In addition to studying PhyAsr, I have developed a simplified methodology to produce a biosensor capable of detecting phosphate release in real-time. The phosphate biosensor is fluorescently labeled and utilizes Escherichia coli Phosphate-binding protein (PhoS). The results show that my proposed methodology yields a functional biosensor and is feasible for large-scale production. I envision this methodology to be versatile and useful for a large number of research applications where detection of free phosphate is required."],"dc:identifier":["hdl:10133/4763"],"dc:subject":["Biochemistry","Enzyme kinetics","Phosphate sensor","PTPLP","myo-inositol","Phosphatase","Stopped-flow","PhyA","PhoS","Phosphate release","Phosphate binding protein","Phytase"],"dc:title":["Rapid kinetic studies of PhyA from Selenomonas ruminantium, and a simplified means of production of a phosphate biosensor"],"dc:type":["Thesis"]},"updated_at":"2026-07-27T20:02:35Z"}