{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/71162"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/71162","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"In Vitro Methanogenesis From Formaldehyde. Identification of Three Carbon-1 Intermediates of the Methanogenic Pathway","abstract":"The biosynthesis of CH(,4) from HCHO and by cell-free extracts of Methanobacterium thermoautotrophicum strain (DELTA)H under H(,2) or N(,2) was investigated. Quantitative conversion of HCHO to CH(,4) was achieved under an atmosphere of H(,2) gas. Under a non-reductive atmosphere of N(,2), HCHO underwent disproportionation (oxidation and reduction) events. A CH(,4)/HCHO ratio of 1/2.9 was observed. Cell-free extracts of methanogens able to oxidize formate to CO(,2) and H(,2) (Mc. jannaschii and Mc. voltae) showed a CH(,4)/HCHO ratio of 1/2. Sets of chemical reactions explaining both findings were proposed. In the case of M. thermoautotrophicum, the oxidized C(,1) units were found bond to a carrier later referred to as Formaldehyde Activating Factor (FAF). A spectrophotometric assay for FAF was implemented and used for the purification of the coenzyme. FAF was purified under strictly anoxic conditions to prevent inactivation by O(,2). ('1)H and ('13)C NMR spectrometry and ultraviolet spectroscopy was used to document the binding of C(,1) units of different oxidation states to FAF. Methanogenesis from each derivative of FAF (formyl, methylene, and methyl) was kinetically comparable to CH(,3)-S-CoM. All the derivatives efficiently substituted for CH(,3)-S-CoM in the CH(,3)-S-CoM-stimulated conversion of CO(,2) to CH(,4), the RPG effect. The enzyme responsible for the oxidation of methylene-FAF to formyl-FAF under N(,2) was found to catalyze the reverse reaction under H(,2). This enzyme was proposed to be part of the methanogenic pathway. Initial purification of the enzyme was reported. Component A(,2) of the methylreductase system was involved in the conversion of HCHO to CH(,4) under H(,2). Initial purification of this enzyme was documented. HCHO, HOCH(,2)-S-CoM, thiazolidine, hexamethylene-tetramine, and L-serine required FAF for their conversion to CH(,4). HCHO was found to react chemically with FAF to form methylene-FAF. HOCH(,2)-S-CoM, thiazolidine, and hexamethylenetetramine were found to be in equilbrium with HCHO. This equilibrium was found to be responsible for the methanogenicity of these compounds. In the case of L-serine, the presence of an FAF-dependent transhydroxymethylase in cell-free extracts of M. thermoautotrophicum was proposed. A new model for methanogenesis from H(,2) and CO(,2) proposed.","abstract_html":"The biosynthesis of CH(,4) from HCHO and by cell-free extracts of Methanobacterium thermoautotrophicum strain (DELTA)H under H(,2) or N(,2) was investigated. Quantitative conversion of HCHO to CH(,4) was achieved under an atmosphere of H(,2) gas. Under a non-reductive atmosphere of N(,2), HCHO underwent disproportionation (oxidation and reduction) events. A CH(,4)/HCHO ratio of 1/2.9 was observed. Cell-free extracts of methanogens able to oxidize formate to CO(,2) and H(,2) (Mc. jannaschii and Mc. voltae) showed a CH(,4)/HCHO ratio of 1/2. Sets of chemical reactions explaining both findings were proposed. In the case of M. thermoautotrophicum, the oxidized C(,1) units were found bond to a carrier later referred to as Formaldehyde Activating Factor (FAF). A spectrophotometric assay for FAF was implemented and used for the purification of the coenzyme. FAF was purified under strictly anoxic conditions to prevent inactivation by O(,2). ('1)H and ('13)C NMR spectrometry and ultraviolet spectroscopy was used to document the binding of C(,1) units of different oxidation states to FAF. Methanogenesis from each derivative of FAF (formyl, methylene, and methyl) was kinetically comparable to CH(,3)-S-CoM. All the derivatives efficiently substituted for CH(,3)-S-CoM in the CH(,3)-S-CoM-stimulated conversion of CO(,2) to CH(,4), the RPG effect. The enzyme responsible for the oxidation of methylene-FAF to formyl-FAF under N(,2) was found to catalyze the reverse reaction under H(,2). This enzyme was proposed to be part of the methanogenic pathway. Initial purification of the enzyme was reported. Component A(,2) of the methylreductase system was involved in the conversion of HCHO to CH(,4) under H(,2). Initial purification of this enzyme was documented. HCHO, HOCH(,2)-S-CoM, thiazolidine, hexamethylene-tetramine, and L-serine required FAF for their conversion to CH(,4). HCHO was found to react chemically with FAF to form methylene-FAF. HOCH(,2)-S-CoM, thiazolidine, and hexamethylenetetramine were found to be in equilbrium with HCHO. This equilibrium was found to be responsible for the methanogenicity of these compounds. In the case of L-serine, the presence of an FAF-dependent transhydroxymethylase in cell-free extracts of M. thermoautotrophicum was proposed. A new model for methanogenesis from H(,2) and CO(,2) proposed.","abstract_has_math":false,"creators":["Escalante Semerena, Jorge Carlos"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Microbiology","degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-12-16T06:12:50Z","date_published":"2014-12-16T06:12:50Z","updated_at":"2026-07-22T22:26:04Z","subjects":["Biology, Microbiology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["(UMI)AAI8409917"],"render_values":[{"text":"(UMI)AAI8409917","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/71162","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Escalante Semerena, Jorge Carlos"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2014-12-16T06:12:50Z","10000-01-01","1983"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Microbiology"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Ph.D."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["University of Illinois at Urbana-Champaign"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biology, Microbiology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["http://hdl.handle.net/2142/71162","(UMI)AAI8409917"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["The biosynthesis of CH(,4) from HCHO and by cell-free extracts of Methanobacterium thermoautotrophicum strain (DELTA)H under H(,2) or N(,2) was investigated. Quantitative conversion of HCHO to CH(,4) was achieved under an atmosphere of H(,2) gas. Under a non-reductive atmosphere of N(,2), HCHO underwent disproportionation (oxidation and reduction) events. A CH(,4)/HCHO ratio of 1/2.9 was observed. Cell-free extracts of methanogens able to oxidize formate to CO(,2) and H(,2) (Mc. jannaschii and Mc. voltae) showed a CH(,4)/HCHO ratio of 1/2. Sets of chemical reactions explaining both findings were proposed. In the case of M. thermoautotrophicum, the oxidized C(,1) units were found bond to a carrier later referred to as Formaldehyde Activating Factor (FAF). A spectrophotometric assay for FAF was implemented and used for the purification of the coenzyme. FAF was purified under strictly anoxic conditions to prevent inactivation by O(,2). ('1)H and ('13)C NMR spectrometry and ultraviolet spectroscopy was used to document the binding of C(,1) units of different oxidation states to FAF. Methanogenesis from each derivative of FAF (formyl, methylene, and methyl) was kinetically comparable to CH(,3)-S-CoM. All the derivatives efficiently substituted for CH(,3)-S-CoM in the CH(,3)-S-CoM-stimulated conversion of CO(,2) to CH(,4), the RPG effect. The enzyme responsible for the oxidation of methylene-FAF to formyl-FAF under N(,2) was found to catalyze the reverse reaction under H(,2). This enzyme was proposed to be part of the methanogenic pathway. Initial purification of the enzyme was reported. Component A(,2) of the methylreductase system was involved in the conversion of HCHO to CH(,4) under H(,2). Initial purification of this enzyme was documented. HCHO, HOCH(,2)-S-CoM, thiazolidine, hexamethylene-tetramine, and L-serine required FAF for their conversion to CH(,4). HCHO was found to react chemically with FAF to form methylene-FAF. HOCH(,2)-S-CoM, thiazolidine, and hexamethylenetetramine were found to be in equilbrium with HCHO. This equilibrium was found to be responsible for the methanogenicity of these compounds. In the case of L-serine, the presence of an FAF-dependent transhydroxymethylase in cell-free extracts of M. thermoautotrophicum was proposed. A new model for methanogenesis from H(,2) and CO(,2) proposed.","Made available in DSpace on 2014-12-16T06:12:50Z (GMT). No. of bitstreams: 1 8409917.pdf: 5273509 bytes, checksum: 2976a3da878e0cccd0463867ec59e40e (MD5) Previous issue date: 1983","Embargo set by: Seth Robbins for item 71328 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","243 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1983."]},{"key":"dc:title","label":"Title","values":["In Vitro Methanogenesis From Formaldehyde. Identification of Three Carbon-1 Intermediates of the Methanogenic Pathway"]}]}],"canonical_facts":{"dc:creator":["Escalante Semerena, Jorge Carlos"],"dc:date":["2014-12-16T06:12:50Z","10000-01-01","1983"],"dc:description":["The biosynthesis of CH(,4) from HCHO and by cell-free extracts of Methanobacterium thermoautotrophicum strain (DELTA)H under H(,2) or N(,2) was investigated. Quantitative conversion of HCHO to CH(,4) was achieved under an atmosphere of H(,2) gas. Under a non-reductive atmosphere of N(,2), HCHO underwent disproportionation (oxidation and reduction) events. A CH(,4)/HCHO ratio of 1/2.9 was observed. Cell-free extracts of methanogens able to oxidize formate to CO(,2) and H(,2) (Mc. jannaschii and Mc. voltae) showed a CH(,4)/HCHO ratio of 1/2. Sets of chemical reactions explaining both findings were proposed. In the case of M. thermoautotrophicum, the oxidized C(,1) units were found bond to a carrier later referred to as Formaldehyde Activating Factor (FAF). A spectrophotometric assay for FAF was implemented and used for the purification of the coenzyme. FAF was purified under strictly anoxic conditions to prevent inactivation by O(,2). ('1)H and ('13)C NMR spectrometry and ultraviolet spectroscopy was used to document the binding of C(,1) units of different oxidation states to FAF. Methanogenesis from each derivative of FAF (formyl, methylene, and methyl) was kinetically comparable to CH(,3)-S-CoM. All the derivatives efficiently substituted for CH(,3)-S-CoM in the CH(,3)-S-CoM-stimulated conversion of CO(,2) to CH(,4), the RPG effect. The enzyme responsible for the oxidation of methylene-FAF to formyl-FAF under N(,2) was found to catalyze the reverse reaction under H(,2). This enzyme was proposed to be part of the methanogenic pathway. Initial purification of the enzyme was reported. Component A(,2) of the methylreductase system was involved in the conversion of HCHO to CH(,4) under H(,2). Initial purification of this enzyme was documented. HCHO, HOCH(,2)-S-CoM, thiazolidine, hexamethylene-tetramine, and L-serine required FAF for their conversion to CH(,4). HCHO was found to react chemically with FAF to form methylene-FAF. HOCH(,2)-S-CoM, thiazolidine, and hexamethylenetetramine were found to be in equilbrium with HCHO. This equilibrium was found to be responsible for the methanogenicity of these compounds. In the case of L-serine, the presence of an FAF-dependent transhydroxymethylase in cell-free extracts of M. thermoautotrophicum was proposed. A new model for methanogenesis from H(,2) and CO(,2) proposed.","Made available in DSpace on 2014-12-16T06:12:50Z (GMT). No. of bitstreams: 1 8409917.pdf: 5273509 bytes, checksum: 2976a3da878e0cccd0463867ec59e40e (MD5) Previous issue date: 1983","Embargo set by: Seth Robbins for item 71328 Lift date: Forever Reason: Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","Restricted to the U of I community idenfinitely during batch ingest of legacy ETDs","U of I Only","243 p.","Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 1983."],"dc:identifier":["http://hdl.handle.net/2142/71162","(UMI)AAI8409917"],"dc:subject":["Biology, Microbiology"],"dc:title":["In Vitro Methanogenesis From Formaldehyde. Identification of Three Carbon-1 Intermediates of the Methanogenic Pathway"],"dc:type":["text"],"thesis:degree_discipline":["Microbiology"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:26:04Z"}