{"id":{"repo_id":"texas-state","oai_identifier":"oai:digital.library.txst.edu:10877/12523"},"canonical_url":"https://search.dev.ndltd.org/etd/texas-state/oai:digital.library.txst.edu:10877/12523","repository":{"repo_id":"texas-state","name":"Texas State University","base_url":"https://digital.library.txst.edu/server/oai/request"},"display":{"title":"Generation and Characterization of dszB Mutants from Nocardia asteroides A3H1","abstract":"Dibenzothiophene, a model compound for sulfur-containing organic molecules found in fossil fuels, can be desulfurized to 2-hydroxybiphenyl by Nocardia asteroides sp. strain A3H1 (A3H1) using the “4S”desulfurization pathway. In the final and rate-limiting step of this pathway, the DszB enzyme catalyzes the hydrolysis of 2-(2’- hydroxyphenyl) benzenesulfinate to form inorganic sulfur and 2-hydroxybiphenyl. The wildtype dszB gene from A3H1 was successfully cloned into the pBAD TOPO® and pBAD TOPO®/ThioFusion expression vectors. Cloning efficiency of the wildtype gene into the pBAD TOPO® vector was 86%. The in vitro directed evolution of DszB was initiated using error-prone PCR. An asexual dszB mutant library was generated and sixty mutant genes were characterized. The average mutation rate of the dszB gene was determined to be 0.75% ± 0.84. Attempts at overexpressing the mutant genes were unsuccessful and no measurable induction of fusion protein expression was observed upon arabinose addition.","abstract_html":"Dibenzothiophene, a model compound for sulfur-containing organic molecules found in fossil fuels, can be desulfurized to 2-hydroxybiphenyl by Nocardia asteroides sp. strain A3H1 (A3H1) using the “4S”desulfurization pathway. In the final and rate-limiting step of this pathway, the DszB enzyme catalyzes the hydrolysis of 2-(2’- hydroxyphenyl) benzenesulfinate to form inorganic sulfur and 2-hydroxybiphenyl. The wildtype dszB gene from A3H1 was successfully cloned into the pBAD TOPO® and pBAD TOPO®/ThioFusion expression vectors. Cloning efficiency of the wildtype gene into the pBAD TOPO® vector was 86%. The in vitro directed evolution of DszB was initiated using error-prone PCR. An asexual dszB mutant library was generated and sixty mutant genes were characterized. The average mutation rate of the dszB gene was determined to be 0.75% ± 0.84. Attempts at overexpressing the mutant genes were unsuccessful and no measurable induction of fusion protein expression was observed upon arabinose addition.","abstract_has_math":false,"creators":["Scott, Christopher A."],"institution":"Southwest Texas State University","degree_name":"Master of Science","degree_level":"Masters","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":["Watkins, Linette M."],"committee_chairs":[],"committee_members":[],"year":2003,"date_issued":"2003-08","date_published":"2003-08","updated_at":"2026-07-27T21:22:51Z","subjects":["desulfurization","Nocardia","petroleum products"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://hdl.handle.net/10877/12523","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.advisor","label":"Advisor","values":["Watkins, Linette M."]},{"key":"dc:creator","label":"Author","values":["Scott, Christopher A."]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2020-09-02T14:36:06Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2020-09-02T14:36:06Z"]},{"key":"dc:date.issued","label":"Date","values":["2003-08"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science"]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Southwest Texas State University"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["desulfurization","Nocardia","petroleum products"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["https://hdl.handle.net/10877/12523"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Dibenzothiophene, a model compound for sulfur-containing organic molecules found in fossil fuels, can be desulfurized to 2-hydroxybiphenyl by Nocardia asteroides sp. strain A3H1 (A3H1) using the “4S”desulfurization pathway. In the final and rate-limiting step of this pathway, the DszB enzyme catalyzes the hydrolysis of 2-(2’- hydroxyphenyl) benzenesulfinate to form inorganic sulfur and 2-hydroxybiphenyl. The wildtype dszB gene from A3H1 was successfully cloned into the pBAD TOPO® and pBAD TOPO®/ThioFusion expression vectors. Cloning efficiency of the wildtype gene into the pBAD TOPO® vector was 86%. The in vitro directed evolution of DszB was initiated using error-prone PCR. An asexual dszB mutant library was generated and sixty mutant genes were characterized. The average mutation rate of the dszB gene was determined to be 0.75% ± 0.84. Attempts at overexpressing the mutant genes were unsuccessful and no measurable induction of fusion protein expression was observed upon arabinose addition."]},{"key":"dc:format","label":"Dc Format","values":["Text"]},{"key":"dc:format.medium","label":"Dc Format Medium","values":["1 file (.pdf)"]},{"key":"dc:title","label":"Title","values":["Generation and Characterization of dszB Mutants from Nocardia asteroides A3H1"]}]}],"canonical_facts":{"dc:contributor.advisor":["Watkins, Linette M."],"dc:creator":["Scott, Christopher A."],"dc:date.accessioned":["2020-09-02T14:36:06Z"],"dc:date.available":["2020-09-02T14:36:06Z"],"dc:date.issued":["2003-08"],"dc:description.abstract":["Dibenzothiophene, a model compound for sulfur-containing organic molecules found in fossil fuels, can be desulfurized to 2-hydroxybiphenyl by Nocardia asteroides sp. strain A3H1 (A3H1) using the “4S”desulfurization pathway. In the final and rate-limiting step of this pathway, the DszB enzyme catalyzes the hydrolysis of 2-(2’- hydroxyphenyl) benzenesulfinate to form inorganic sulfur and 2-hydroxybiphenyl. The wildtype dszB gene from A3H1 was successfully cloned into the pBAD TOPO® and pBAD TOPO®/ThioFusion expression vectors. Cloning efficiency of the wildtype gene into the pBAD TOPO® vector was 86%. The in vitro directed evolution of DszB was initiated using error-prone PCR. An asexual dszB mutant library was generated and sixty mutant genes were characterized. The average mutation rate of the dszB gene was determined to be 0.75% ± 0.84. Attempts at overexpressing the mutant genes were unsuccessful and no measurable induction of fusion protein expression was observed upon arabinose addition."],"dc:format":["Text"],"dc:format.medium":["1 file (.pdf)"],"dc:identifier.uri":["https://hdl.handle.net/10877/12523"],"dc:language.iso":["en"],"dc:subject":["desulfurization","Nocardia","petroleum products"],"dc:title":["Generation and Characterization of dszB Mutants from Nocardia asteroides A3H1"],"dc:type":["Thesis"],"thesis:degree_level":["Masters"],"thesis:degree_name":["Master of Science"],"thesis:institution_name":["Southwest Texas State University"]},"updated_at":"2026-07-27T21:22:51Z"}