Isolation, characterization, and pectin-modifying properties of a thermally tolerant pectin methylesterase from Citrus sinensis var. Valencia.
Identifieur interne : 002825 ( Main/Merge ); précédent : 002824; suivant : 002826Isolation, characterization, and pectin-modifying properties of a thermally tolerant pectin methylesterase from Citrus sinensis var. Valencia.
Auteurs : Randall G. Cameron [États-Unis] ; Brett J. Savary ; Arland T. Hotchkiss ; Marshall L. FishmanSource :
- Journal of agricultural and food chemistry [ 0021-8561 ] ; 2005.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : Carboxylic Ester Hydrolases.
- chemical , metabolism : Carboxylic Ester Hydrolases, Pectins.
- chemistry : Plant Leaves.
- enzymology : Citrus sinensis, Fruit.
- Hot Temperature.
Abstract
The thermally tolerant pectin methylesterase (TT-PME) was isolated as a monocomponent enzyme from sweet orange fruit (Citrus sinensis var. Valencia). It was also isolated from flower and vegetative tissue. The apparent molecular weight of fruit TT-PME was 40800 by SDS-PAGE and the isoelectric point estimated as pI 9.31 by IEF-PAGE. MALDI-TOF MS identified no tryptic-peptide ions from TT-PME characteristic of previously described citrus PMEs. TT-PME did not absolutely require supplemented salt for activity, but salt activation and pH-dependent activity patterns were intermediate to those of thermolabile PMEs. Treatment of non-calcium-sensitive pectin with TT-PME (reducing the degree of methylesterification by 6%) increased the calcium-sensitive pectin ratio from 0.01 to 0.90, indicating a blockwise mode of action. TT-PME produced a significantly lower end-point degree of methylesterification at pH 7.5 than at pH 4.5. Extensive de-esterification with TT-PME did not reduce the pectin molecular weight or z-average radius of gyration, as determined by HPSEC.
DOI: 10.1021/jf040376m
PubMed: 15769165
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pubmed:15769165Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Isolation, characterization, and pectin-modifying properties of a thermally tolerant pectin methylesterase from Citrus sinensis var. Valencia.</title><author><name sortKey="Cameron, Randall G" sort="Cameron, Randall G" uniqKey="Cameron R" first="Randall G" last="Cameron">Randall G. Cameron</name><affiliation wicri:level="1"><nlm:affiliation>Quality Improvement in Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S N.W., Winter Haven, Florida 33881, USA. rcameron@citrus.usda.gov</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Quality Improvement in Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S N.W., Winter Haven, Florida 33881</wicri:regionArea><wicri:noRegion>Florida 33881</wicri:noRegion></affiliation></author><author><name sortKey="Savary, Brett J" sort="Savary, Brett J" uniqKey="Savary B" first="Brett J" last="Savary">Brett J. Savary</name></author><author><name sortKey="Hotchkiss, Arland T" sort="Hotchkiss, Arland T" uniqKey="Hotchkiss A" first="Arland T" last="Hotchkiss">Arland T. Hotchkiss</name></author><author><name sortKey="Fishman, Marshall L" sort="Fishman, Marshall L" uniqKey="Fishman M" first="Marshall L" last="Fishman">Marshall L. Fishman</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2005">2005</date><idno type="RBID">pubmed:15769165</idno><idno type="pmid">15769165</idno><idno type="doi">10.1021/jf040376m</idno><idno type="wicri:Area/PubMed/Corpus">000D19</idno><idno type="wicri:Area/PubMed/Curation">000D19</idno><idno type="wicri:Area/PubMed/Checkpoint">000D19</idno><idno type="wicri:Area/Ncbi/Merge">000367</idno><idno type="wicri:Area/Ncbi/Curation">000367</idno><idno type="wicri:Area/Ncbi/Checkpoint">000367</idno><idno type="wicri:doubleKey">0021-8561:2005:Cameron R:isolation:characterization:and</idno><idno type="wicri:Area/Main/Merge">002825</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Isolation, characterization, and pectin-modifying properties of a thermally tolerant pectin methylesterase from Citrus sinensis var. Valencia.</title><author><name sortKey="Cameron, Randall G" sort="Cameron, Randall G" uniqKey="Cameron R" first="Randall G" last="Cameron">Randall G. Cameron</name><affiliation wicri:level="1"><nlm:affiliation>Quality Improvement in Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S N.W., Winter Haven, Florida 33881, USA. rcameron@citrus.usda.gov</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Quality Improvement in Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S N.W., Winter Haven, Florida 33881</wicri:regionArea><wicri:noRegion>Florida 33881</wicri:noRegion></affiliation></author><author><name sortKey="Savary, Brett J" sort="Savary, Brett J" uniqKey="Savary B" first="Brett J" last="Savary">Brett J. Savary</name></author><author><name sortKey="Hotchkiss, Arland T" sort="Hotchkiss, Arland T" uniqKey="Hotchkiss A" first="Arland T" last="Hotchkiss">Arland T. Hotchkiss</name></author><author><name sortKey="Fishman, Marshall L" sort="Fishman, Marshall L" uniqKey="Fishman M" first="Marshall L" last="Fishman">Marshall L. Fishman</name></author></analytic><series><title level="j">Journal of agricultural and food chemistry</title><idno type="ISSN">0021-8561</idno><imprint><date when="2005" type="published">2005</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Carboxylic Ester Hydrolases (isolation & purification)</term><term>Carboxylic Ester Hydrolases (metabolism)</term><term>Citrus sinensis (enzymology)</term><term>Fruit (enzymology)</term><term>Hot Temperature</term><term>Pectins (metabolism)</term><term>Plant Leaves (chemistry)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Carboxylic Ester Hydrolases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Carboxylic Ester Hydrolases</term><term>Pectins</term></keywords><keywords scheme="MESH" qualifier="chemistry" xml:lang="en"><term>Plant Leaves</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Citrus sinensis</term><term>Fruit</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Hot Temperature</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">The thermally tolerant pectin methylesterase (TT-PME) was isolated as a monocomponent enzyme from sweet orange fruit (Citrus sinensis var. Valencia). It was also isolated from flower and vegetative tissue. The apparent molecular weight of fruit TT-PME was 40800 by SDS-PAGE and the isoelectric point estimated as pI 9.31 by IEF-PAGE. MALDI-TOF MS identified no tryptic-peptide ions from TT-PME characteristic of previously described citrus PMEs. TT-PME did not absolutely require supplemented salt for activity, but salt activation and pH-dependent activity patterns were intermediate to those of thermolabile PMEs. Treatment of non-calcium-sensitive pectin with TT-PME (reducing the degree of methylesterification by 6%) increased the calcium-sensitive pectin ratio from 0.01 to 0.90, indicating a blockwise mode of action. TT-PME produced a significantly lower end-point degree of methylesterification at pH 7.5 than at pH 4.5. Extensive de-esterification with TT-PME did not reduce the pectin molecular weight or z-average radius of gyration, as determined by HPSEC.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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