Purification and characterization of a beta-glucosidase from Citrus sinensis var. Valencia fruit tissue.
Identifieur interne : 000E29 ( PubMed/Checkpoint ); précédent : 000E28; suivant : 000E30Purification and characterization of a beta-glucosidase from Citrus sinensis var. Valencia fruit tissue.
Auteurs : R G Cameron [États-Unis] ; J A Manthey ; R A Baker ; K. GrohmannSource :
- Journal of agricultural and food chemistry [ 0021-8561 ] ; 2001.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : beta-Glucosidase.
- enzymology : Citrus.
- chemical , metabolism : beta-Glucosidase.
- Chromatography, Gel, Hydrogen-Ion Concentration, Hydrolysis, Kinetics, Molecular Weight, Temperature.
Abstract
A preliminary survey demonstrated activity for alpha-D-glucosidase, alpha-D-mannosidase, alpha-L-arabinosidase, beta-D-glucosidase, beta-D-xylosidase, and beta-D-galactosidase in orange fruit flavedo and albedo tissue. alpha-L-Rhamnosidase was not detected. Subsequently, a beta-glucosidase was purified from mature fruit rag tissue (composed of intersegmental septa, squeezed juice sacs, and fruit core tissue) of Citrus sinensis var. Valencia. The beta-glucosidase exhibited low levels of activity against p-nitrophenyl-beta-D-fucopyranoside (13.5%) and p-nitrophenyl-alpha-D-glucopyranoside (7.0%), compared to its activity against p-nitrophenyl-beta-D-glucopyranoside (pNPG, 100%). The enzyme was purified by a combination of ion exchange (anion and cation) and gel filtration (Superdex and Toyopearl HW-55S) chromatography. It has an apparent molecular mass of 64 kDa by denaturing electrophoresis or 55 kDa by gel filtration chromatography (BioGel P-100). Hydrolysis of pNPG demonstrated a pH optimum between 4.5 and 5.5. At pH 5.0 the temperature optimum was 40 degrees C. At pH 5.0 and 40 degrees C the K(m) for pNPG was 0.1146 mM and it had a V(max) of 5.2792 nkatal x mg(-1) protein (katal = 0.06 International Units = the amount of enzyme that produces, under standard conditions, one micromol of product per min). Of the substrates tested, the enzyme was most active against the disaccharide cellobiose (1-->4), but was not active against p-nitrophenyl-beta-D-cellobioside. High levels of activity also were observed with the disaccharides laminaribiose (1-->3), gentiobiose (1-->6), and sophorose (1-->2). Activity greater than that observed with pNPG was obtained with the flavonoids hesperetin-7-glucoside and prunin (naringenin-7-glucoside), salicin, mandelonitrile-beta-D-glucoside (a cyanogenic substrate), and sinigrin (a glucosinolate). The enzyme was not active against amygdalin, coniferin, or limonin glucoside.
PubMed: 11559154
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Valencia fruit tissue.</title><author><name sortKey="Cameron, R G" sort="Cameron, R G" uniqKey="Cameron R" first="R G" last="Cameron">R G Cameron</name><affiliation wicri:level="1"><nlm:affiliation>Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S, NW, Winter Haven, Florida 33881, USA. rcameron@citrus.usda.gov</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S, NW, Winter Haven, Florida 33881</wicri:regionArea><wicri:noRegion>Florida 33881</wicri:noRegion></affiliation></author><author><name sortKey="Manthey, J A" sort="Manthey, J A" uniqKey="Manthey J" first="J A" last="Manthey">J A Manthey</name></author><author><name sortKey="Baker, R A" sort="Baker, R A" uniqKey="Baker R" first="R A" last="Baker">R A Baker</name></author><author><name sortKey="Grohmann, K" sort="Grohmann, K" uniqKey="Grohmann K" first="K" last="Grohmann">K. 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Valencia fruit tissue.</title><author><name sortKey="Cameron, R G" sort="Cameron, R G" uniqKey="Cameron R" first="R G" last="Cameron">R G Cameron</name><affiliation wicri:level="1"><nlm:affiliation>Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S, NW, Winter Haven, Florida 33881, USA. rcameron@citrus.usda.gov</nlm:affiliation><country xml:lang="fr">États-Unis</country><wicri:regionArea>Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S, NW, Winter Haven, Florida 33881</wicri:regionArea><wicri:noRegion>Florida 33881</wicri:noRegion></affiliation></author><author><name sortKey="Manthey, J A" sort="Manthey, J A" uniqKey="Manthey J" first="J A" last="Manthey">J A Manthey</name></author><author><name sortKey="Baker, R A" sort="Baker, R A" uniqKey="Baker R" first="R A" last="Baker">R A Baker</name></author><author><name sortKey="Grohmann, K" sort="Grohmann, K" uniqKey="Grohmann K" first="K" last="Grohmann">K. Grohmann</name></author></analytic><series><title level="j">Journal of agricultural and food chemistry</title><idno type="ISSN">0021-8561</idno><imprint><date when="2001" type="published">2001</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Chromatography, Gel</term><term>Citrus (enzymology)</term><term>Hydrogen-Ion Concentration</term><term>Hydrolysis</term><term>Kinetics</term><term>Molecular Weight</term><term>Temperature</term><term>beta-Glucosidase (isolation & purification)</term><term>beta-Glucosidase (metabolism)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>beta-Glucosidase</term></keywords><keywords scheme="MESH" qualifier="enzymology" xml:lang="en"><term>Citrus</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>beta-Glucosidase</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Chromatography, Gel</term><term>Hydrogen-Ion Concentration</term><term>Hydrolysis</term><term>Kinetics</term><term>Molecular Weight</term><term>Temperature</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A preliminary survey demonstrated activity for alpha-D-glucosidase, alpha-D-mannosidase, alpha-L-arabinosidase, beta-D-glucosidase, beta-D-xylosidase, and beta-D-galactosidase in orange fruit flavedo and albedo tissue. alpha-L-Rhamnosidase was not detected. Subsequently, a beta-glucosidase was purified from mature fruit rag tissue (composed of intersegmental septa, squeezed juice sacs, and fruit core tissue) of Citrus sinensis var. Valencia. The beta-glucosidase exhibited low levels of activity against p-nitrophenyl-beta-D-fucopyranoside (13.5%) and p-nitrophenyl-alpha-D-glucopyranoside (7.0%), compared to its activity against p-nitrophenyl-beta-D-glucopyranoside (pNPG, 100%). The enzyme was purified by a combination of ion exchange (anion and cation) and gel filtration (Superdex and Toyopearl HW-55S) chromatography. It has an apparent molecular mass of 64 kDa by denaturing electrophoresis or 55 kDa by gel filtration chromatography (BioGel P-100). Hydrolysis of pNPG demonstrated a pH optimum between 4.5 and 5.5. At pH 5.0 the temperature optimum was 40 degrees C. At pH 5.0 and 40 degrees C the K(m) for pNPG was 0.1146 mM and it had a V(max) of 5.2792 nkatal x mg(-1) protein (katal = 0.06 International Units = the amount of enzyme that produces, under standard conditions, one micromol of product per min). Of the substrates tested, the enzyme was most active against the disaccharide cellobiose (1-->4), but was not active against p-nitrophenyl-beta-D-cellobioside. High levels of activity also were observed with the disaccharides laminaribiose (1-->3), gentiobiose (1-->6), and sophorose (1-->2). Activity greater than that observed with pNPG was obtained with the flavonoids hesperetin-7-glucoside and prunin (naringenin-7-glucoside), salicin, mandelonitrile-beta-D-glucoside (a cyanogenic substrate), and sinigrin (a glucosinolate). The enzyme was not active against amygdalin, coniferin, or limonin glucoside.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">11559154</PMID><DateCreated><Year>2001</Year><Month>9</Month><Day>17</Day></DateCreated><DateCompleted><Year>2001</Year><Month>10</Month><Day>25</Day></DateCompleted><DateRevised><Year>2001</Year><Month>9</Month><Day>17</Day></DateRevised><Article PubModel="Print"><Journal><ISSN IssnType="Print">0021-8561</ISSN><JournalIssue CitedMedium="Print"><Volume>49</Volume><Issue>9</Issue><PubDate><Year>2001</Year><Month>Sep</Month></PubDate></JournalIssue><Title>Journal of agricultural and food chemistry</Title><ISOAbbreviation>J. Agric. Food Chem.</ISOAbbreviation></Journal><ArticleTitle>Purification and characterization of a beta-glucosidase from Citrus sinensis var. Valencia fruit tissue.</ArticleTitle><Pagination><MedlinePgn>4457-62</MedlinePgn></Pagination><Abstract><AbstractText>A preliminary survey demonstrated activity for alpha-D-glucosidase, alpha-D-mannosidase, alpha-L-arabinosidase, beta-D-glucosidase, beta-D-xylosidase, and beta-D-galactosidase in orange fruit flavedo and albedo tissue. alpha-L-Rhamnosidase was not detected. Subsequently, a beta-glucosidase was purified from mature fruit rag tissue (composed of intersegmental septa, squeezed juice sacs, and fruit core tissue) of Citrus sinensis var. Valencia. The beta-glucosidase exhibited low levels of activity against p-nitrophenyl-beta-D-fucopyranoside (13.5%) and p-nitrophenyl-alpha-D-glucopyranoside (7.0%), compared to its activity against p-nitrophenyl-beta-D-glucopyranoside (pNPG, 100%). The enzyme was purified by a combination of ion exchange (anion and cation) and gel filtration (Superdex and Toyopearl HW-55S) chromatography. It has an apparent molecular mass of 64 kDa by denaturing electrophoresis or 55 kDa by gel filtration chromatography (BioGel P-100). Hydrolysis of pNPG demonstrated a pH optimum between 4.5 and 5.5. At pH 5.0 the temperature optimum was 40 degrees C. At pH 5.0 and 40 degrees C the K(m) for pNPG was 0.1146 mM and it had a V(max) of 5.2792 nkatal x mg(-1) protein (katal = 0.06 International Units = the amount of enzyme that produces, under standard conditions, one micromol of product per min). Of the substrates tested, the enzyme was most active against the disaccharide cellobiose (1-->4), but was not active against p-nitrophenyl-beta-D-cellobioside. High levels of activity also were observed with the disaccharides laminaribiose (1-->3), gentiobiose (1-->6), and sophorose (1-->2). Activity greater than that observed with pNPG was obtained with the flavonoids hesperetin-7-glucoside and prunin (naringenin-7-glucoside), salicin, mandelonitrile-beta-D-glucoside (a cyanogenic substrate), and sinigrin (a glucosinolate). The enzyme was not active against amygdalin, coniferin, or limonin glucoside.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Cameron</LastName><ForeName>R G</ForeName><Initials>RG</Initials><AffiliationInfo><Affiliation>Citrus and Subtropical Products Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 600 Avenue S, NW, Winter Haven, Florida 33881, USA. rcameron@citrus.usda.gov</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Manthey</LastName><ForeName>J A</ForeName><Initials>JA</Initials></Author><Author ValidYN="Y"><LastName>Baker</LastName><ForeName>R A</ForeName><Initials>RA</Initials></Author><Author ValidYN="Y"><LastName>Grohmann</LastName><ForeName>K</ForeName><Initials>K</Initials></Author></AuthorList><Language>ENG</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>J Agric Food Chem</MedlineTA><NlmUniqueID>0374755</NlmUniqueID><ISSNLinking>0021-8561</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>EC 3.2.1.21</RegistryNumber><NameOfSubstance UI="D001617">beta-Glucosidase</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D002850" MajorTopicYN="N">Chromatography, Gel</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D002957" MajorTopicYN="N">Citrus</DescriptorName><QualifierName UI="Q000201" MajorTopicYN="Y">enzymology</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006863" MajorTopicYN="N">Hydrogen-Ion Concentration</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006868" MajorTopicYN="N">Hydrolysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D007700" MajorTopicYN="N">Kinetics</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008970" MajorTopicYN="N">Molecular Weight</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013696" MajorTopicYN="N">Temperature</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001617" MajorTopicYN="N">beta-Glucosidase</DescriptorName><QualifierName UI="Q000302" MajorTopicYN="Y">isolation & purification</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="pubmed"><Year>2001</Year><Month>9</Month><Day>18</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2001</Year><Month>10</Month><Day>26</Day><Hour>10</Hour><Minute>1</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2001</Year><Month>9</Month><Day>18</Day><Hour>10</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">11559154</ArticleId><ArticleId IdType="pii">jf010010z</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Baker, R A" sort="Baker, R A" uniqKey="Baker R" first="R A" last="Baker">R A Baker</name><name sortKey="Grohmann, K" sort="Grohmann, K" uniqKey="Grohmann K" first="K" last="Grohmann">K. Grohmann</name><name sortKey="Manthey, J A" sort="Manthey, J A" uniqKey="Manthey J" first="J A" last="Manthey">J A Manthey</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Cameron, R G" sort="Cameron, R G" uniqKey="Cameron R" first="R G" last="Cameron">R G Cameron</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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