Structural Characterization of the Thermally Tolerant Pectin Methylesterase Purified from Citrus sinensis Fruit and Its Gene Sequence
Identifieur interne : 001113 ( Main/Curation ); précédent : 001112; suivant : 001114Structural Characterization of the Thermally Tolerant Pectin Methylesterase Purified from Citrus sinensis Fruit and Its Gene Sequence
Auteurs : Brett J. Savary [États-Unis] ; Prasanna Vasu [États-Unis, Inde] ; Randall G. Cameron [États-Unis] ; T. Gregory Mccollum [États-Unis] ; Alberto Nunez [États-Unis]Source :
- Journal of agricultural and food chemistry : (Print) [ 0021-8561 ] ; 2013.
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- topic : Fruit.
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Abstract
Despite the longstanding importance of the thermally tolerant pectin methylesterase (TT-PME) activity in citrus juice processing and product quality, the unequivocal identification of the protein and its corresponding gene has remained elusive. TT-PME was purified from sweet orange [Citrus sinensis (L.) Osbeck] finisher pulp (8.0 mg/1.3 kg tissue) with an improved purification scheme that provided 20-fold increased enzyme yield over previous results. Structural characterization of electrophoretically pure TT-PME by MALDI-TOF MS determined molecular masses of approximately 47900 and 53000 Da for two principal glycoisoforms. De novo sequences generated from tryptic peptides by MALDI-TOF/TOF MS matched multiple anonymous Citrus EST cDNA accessions. The complete tt-pme cDNA (1710 base pair) was cloned from a fruit mRNA library using RT- and RLM-RACE PCR. Citrus TT-PME is a novel isoform that showed higher sequence identity with the multiply glycosylated kiwifruit PME than to previously described Citrus thermally labile PME isoforms.
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Savary</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor</s1><s2>Pennsylvania 19038</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Pennsylvania 19038</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Arkansas Biosciences Institute, Arkansas State University, P.O. Box 639, State University</s1><s2>Arkansas 72467</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Arkansas 72467</wicri:noRegion></affiliation></author><author><name sortKey="Vasu, Prasanna" sort="Vasu, Prasanna" uniqKey="Vasu P" first="Prasanna" last="Vasu">Prasanna Vasu</name><affiliation wicri:level="1"><inist:fA14 i1="02"><s1>Arkansas Biosciences Institute, Arkansas State University, P.O. Box 639, State University</s1><s2>Arkansas 72467</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>2 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Arkansas 72467</wicri:noRegion></affiliation><affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Food Safety and Analytical Quality Control Laboratory, Central Food Technological Research Institute, CSIR</s1><s2>Mysore 570 020</s2><s3>IND</s3><sZ>2 aut.</sZ></inist:fA14><country>Inde</country><wicri:noRegion>Mysore 570 020</wicri:noRegion></affiliation></author><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"><inist:fA14 i1="04"><s1>U.S. Horticultural Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 2001 South Rock Road</s1><s2>Fort Pierce, Florida 34945</s2><s3>USA</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Fort Pierce, Florida 34945</wicri:noRegion></affiliation></author><author><name sortKey="Mccollum, T Gregory" sort="Mccollum, T Gregory" uniqKey="Mccollum T" first="T. Gregory" last="Mccollum">T. Gregory Mccollum</name><affiliation wicri:level="1"><inist:fA14 i1="04"><s1>U.S. Horticultural Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 2001 South Rock Road</s1><s2>Fort Pierce, Florida 34945</s2><s3>USA</s3><sZ>3 aut.</sZ><sZ>4 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Fort Pierce, Florida 34945</wicri:noRegion></affiliation></author><author><name sortKey="Nunez, Alberto" sort="Nunez, Alberto" uniqKey="Nunez A" first="Alberto" last="Nunez">Alberto Nunez</name><affiliation wicri:level="1"><inist:fA14 i1="01"><s1>Eastern Regional Research Center, Agricultural Research Service, U.S. Department of Agriculture, Wyndmoor</s1><s2>Pennsylvania 19038</s2><s3>USA</s3><sZ>1 aut.</sZ><sZ>5 aut.</sZ></inist:fA14><country>États-Unis</country><wicri:noRegion>Pennsylvania 19038</wicri:noRegion></affiliation></author></analytic><series><title level="j" type="main">Journal of agricultural and food chemistry : (Print)</title><title level="j" type="abbreviated">J. agric. food chem. : (Print)</title><idno type="ISSN">0021-8561</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc><seriesStmt><title level="j" type="main">Journal of agricultural and food chemistry : (Print)</title><title level="j" type="abbreviated">J. agric. food chem. : (Print)</title><idno type="ISSN">0021-8561</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Cell wall</term><term>Characterization</term><term>Citrus sinensis</term><term>Fruit</term><term>Matrix assisted laser desorption ionization</term><term>Nucleotide sequence</term><term>Orange juice</term><term>Pectinesterase</term><term>Polymerase chain reaction</term><term>Purification</term><term>Thermal stability</term><term>Tolerance</term></keywords><keywords scheme="Pascal" xml:lang="fr"><term>Caractérisation</term><term>Tolérance</term><term>Pectinesterase</term><term>Fruit</term><term>Séquence nucléotide</term><term>Citrus sinensis</term><term>Paroi cellulaire</term><term>Jus d'orange</term><term>Stabilité thermique</term><term>MALDI</term><term>Réaction chaîne polymérase</term><term>Purification</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Fruit</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Despite the longstanding importance of the thermally tolerant pectin methylesterase (TT-PME) activity in citrus juice processing and product quality, the unequivocal identification of the protein and its corresponding gene has remained elusive. TT-PME was purified from sweet orange [Citrus sinensis (L.) Osbeck] finisher pulp (8.0 mg/1.3 kg tissue) with an improved purification scheme that provided 20-fold increased enzyme yield over previous results. Structural characterization of electrophoretically pure TT-PME by MALDI-TOF MS determined molecular masses of approximately 47900 and 53000 Da for two principal glycoisoforms. De novo sequences generated from tryptic peptides by MALDI-TOF/TOF MS matched multiple anonymous Citrus EST cDNA accessions. The complete tt-pme cDNA (1710 base pair) was cloned from a fruit mRNA library using RT- and RLM-RACE PCR. Citrus TT-PME is a novel isoform that showed higher sequence identity with the multiply glycosylated kiwifruit PME than to previously described Citrus thermally labile PME isoforms.</div></front></TEI></record>Pour manipuler ce document sous Unix (Dilib)
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