{"id":{"repo_id":"uiuc","oai_identifier":"oai:www.ideals.illinois.edu:2142/19012"},"canonical_url":"https://search.dev.ndltd.org/etd/uiuc/oai:www.ideals.illinois.edu:2142/19012","repository":{"repo_id":"uiuc","name":"University of Illinois - Urbana-Champaign","base_url":"https://www.ideals.illinois.edu/oai-pmh"},"display":{"title":"Isolation and characterization of carotenoproteins in higher plant systems","abstract":"For the first time, a procedure has been developed for the isolation of intact $\\alpha$- and $\\beta$-carotene binding carotenoprotein from carrot chromoplasts. Initial attempts to isolate and purify this protein from spinach chloroplasts was unsuccessful because of contaminating chlorophyll and chlorophyll proteins.","abstract_html":"For the first time, a procedure has been developed for the isolation of intact <span class=\"etd-inline-math\">&alpha;</span>- and <span class=\"etd-inline-math\">&beta;</span>-carotene binding carotenoprotein from carrot chromoplasts. Initial attempts to isolate and purify this protein from spinach chloroplasts was unsuccessful because of contaminating chlorophyll and chlorophyll proteins.","abstract_has_math":true,"creators":["Dietz, Jane Marie"],"institution":"University of Illinois at Urbana-Champaign","degree_name":"Ph.D.","degree_level":"Dissertation","degree_discipline":"Food Sciences","degree_department":null,"school":null,"contributors":["Erdman, John W."],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2011,"date_issued":"2011-05-07T11:54:18Z","date_published":"2011-05-07T11:54:18Z","updated_at":"2026-07-22T22:25:12Z","subjects":["Agriculture","Health Sciences","Food Science","Technology","Nutrition"],"languages":["eng"],"rights":["Copyright 1990 Dietz, Jane Marie"],"rights_urls":[],"identifier_entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9114221","(UMI)AAI9114221"],"render_values":[{"text":"AAI9114221","href":null,"code":true},{"text":"(UMI)AAI9114221","href":null,"code":true}]}]},"links":{"outbound_url":"http://hdl.handle.net/2142/19012","outbound_label":"Handle","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Erdman, John W."]},{"key":"dc:creator","label":"Author","values":["Dietz, Jane Marie"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2011-05-07T11:54:18Z","10000-01-01","1990"]},{"key":"dc:type","label":"Dc Type","values":["text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Food Sciences"]},{"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":["Agriculture","Health Sciences","Food Science","Technology","Nutrition"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language","label":"Dc Language","values":["eng"]},{"key":"dc:rights","label":"Dc Rights","values":["Copyright 1990 Dietz, Jane Marie"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["AAI9114221","(UMI)AAI9114221","http://hdl.handle.net/2142/19012"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["For the first time, a procedure has been developed for the isolation of intact $\\alpha$- and $\\beta$-carotene binding carotenoprotein from carrot chromoplasts. Initial attempts to isolate and purify this protein from spinach chloroplasts was unsuccessful because of contaminating chlorophyll and chlorophyll proteins.","Though the attempts were unsuccessful for a carotene binding protein in spinach, a lutein isomer binding carotenoprotein was effectively isolated. The isolation method employed metal chelate affinity chromatography using immobilized copper ions. While suitable for this more polar carotenoid protein complex, this method was not utilizable for non-polar carotene binding proteins. Degradation of $\\beta$-carotene occurred during chromatography, apparently catalyzed by the copper ions.","The developed procedure for the isolation of the pure carotenoprotein from carrot chromoplast was quick and straightforward. Only French pressing, high speed centrifugation and gel filtration were needed to purify the protein.","The major protein peak collected from the gel filtration column was also the major carotenoid peak. This peak corresponded to a molecular weight of approximately 2,000 kDal for the native protein. Isoelectric focusing indicated the presence of a single protein band with a pI of 3.6. Further substantiation that this was a single pure protein complex was demonstrated by SDS-PAGE where only a single 54 kDal protein band was detected. This indicates that the native complex of 2,000 kDal was comprised of seemingly identical 54 kDal subunits.","Analysis of the carotenoprotein complex by r-phase HPLC demonstrated the protein to bind 1 mole of $\\alpha$-carotene and 2 moles of $\\beta$-carotene for each mole of the subunit.","The amino acid composition of the protein suggests that the protein contains a high percentage of $\\alpha$-helical conformation in combination with a probability of numerous reverse turns. Given this information and the high percentage of hydrophobic amino acids present, it may be hypothesized that the helical structure of the protein may form a hydrophobic cleft along the surface of the protein which would allow attachment of the carotenes. This hypothesis is consistent with the ease in which the carotene may be dissociated from the protein complex during the isolation procedures.","Made available in DSpace on 2011-05-07T11:54:18Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9114221.pdf: 4916615 bytes, checksum: d728e1adf983420747b0e951bc91e92b (MD5) Previous issue date: 1990","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:34:03Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:12:50-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"]},{"key":"dc:title","label":"Title","values":["Isolation and characterization of carotenoproteins in higher plant systems"]}]}],"canonical_facts":{"dc:contributor":["Erdman, John W."],"dc:creator":["Dietz, Jane Marie"],"dc:date":["2011-05-07T11:54:18Z","10000-01-01","1990"],"dc:description":["For the first time, a procedure has been developed for the isolation of intact $\\alpha$- and $\\beta$-carotene binding carotenoprotein from carrot chromoplasts. Initial attempts to isolate and purify this protein from spinach chloroplasts was unsuccessful because of contaminating chlorophyll and chlorophyll proteins.","Though the attempts were unsuccessful for a carotene binding protein in spinach, a lutein isomer binding carotenoprotein was effectively isolated. The isolation method employed metal chelate affinity chromatography using immobilized copper ions. While suitable for this more polar carotenoid protein complex, this method was not utilizable for non-polar carotene binding proteins. Degradation of $\\beta$-carotene occurred during chromatography, apparently catalyzed by the copper ions.","The developed procedure for the isolation of the pure carotenoprotein from carrot chromoplast was quick and straightforward. Only French pressing, high speed centrifugation and gel filtration were needed to purify the protein.","The major protein peak collected from the gel filtration column was also the major carotenoid peak. This peak corresponded to a molecular weight of approximately 2,000 kDal for the native protein. Isoelectric focusing indicated the presence of a single protein band with a pI of 3.6. Further substantiation that this was a single pure protein complex was demonstrated by SDS-PAGE where only a single 54 kDal protein band was detected. This indicates that the native complex of 2,000 kDal was comprised of seemingly identical 54 kDal subunits.","Analysis of the carotenoprotein complex by r-phase HPLC demonstrated the protein to bind 1 mole of $\\alpha$-carotene and 2 moles of $\\beta$-carotene for each mole of the subunit.","The amino acid composition of the protein suggests that the protein contains a high percentage of $\\alpha$-helical conformation in combination with a probability of numerous reverse turns. Given this information and the high percentage of hydrophobic amino acids present, it may be hypothesized that the helical structure of the protein may form a hydrophobic cleft along the surface of the protein which would allow attachment of the carotenes. This hypothesis is consistent with the ease in which the carotene may be dissociated from the protein complex during the isolation procedures.","Made available in DSpace on 2011-05-07T11:54:18Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9114221.pdf: 4916615 bytes, checksum: d728e1adf983420747b0e951bc91e92b (MD5) Previous issue date: 1990","Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T14:34:03Z Item is restricted indefinitely.","Restriction data tranferred 2014-07-01T11:12:50-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission","ETDs are only available to UIUC Users without author permission","U of I Only"],"dc:identifier":["AAI9114221","(UMI)AAI9114221","http://hdl.handle.net/2142/19012"],"dc:language":["eng"],"dc:rights":["Copyright 1990 Dietz, Jane Marie"],"dc:subject":["Agriculture","Health Sciences","Food Science","Technology","Nutrition"],"dc:title":["Isolation and characterization of carotenoproteins in higher plant systems"],"dc:type":["text"],"thesis:degree_discipline":["Food Sciences"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Ph.D."],"thesis:institution_name":["University of Illinois at Urbana-Champaign"]},"updated_at":"2026-07-22T22:25:12Z"}