Purification and characterization of a beta-glucosidase from Citrus sinensis var. valencia fruit tissue
Identifieur interne : 002E11 ( Main/Exploration ); précédent : 002E10; suivant : 002E12Purification and characterization of a beta-glucosidase from Citrus sinensis var. valencia fruit tissue
Auteurs : Randall G. Cameron [États-Unis] ; John A. Manthey [États-Unis] ; Robert A. Baker [États-Unis] ; Karel Grohmann [États-Unis]Source :
- Journal of agricultural and food chemistry : (Print) [ 0021-8561 ] ; 2001.
Descripteurs français
- Pascal (Inist)
- Wicri :
- topic : Fruit.
English descriptors
- KwdEn :
- By product, Catalyst activity, Characterization, Chromatography, Gel, Citrus (enzymology), Citrus sinensis, Enzymatic activity, Fruit, Hydrogen-Ion Concentration, Hydrolysis, Kinetics, Molecular Weight, Molecular mass, Optimum pH, Orange juice, Purification, Substrate specificity, Temperature, Tissue specificity, beta-Glucosidase (isolation & purification), beta-Glucosidase (metabolism), β-Glucosidase.
- 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 α-D-glucosidase, α-D-mannosidase, α-L-arabinosidase, β-D-glucosidase, β-D-xylosidase, and β-D-galactosidase in orange fruit flavedo and albedo tissue, α-L-Rhamnosidase was not detected. Subsequently, a β-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 β-glucosidase exhibited low levels of activity against p-nitrophenyl-β-D-fucopyranoside (13.5%) and p-nitrophenyl-a-D-glucopyranoside (7.0%), compared to its activity against p-nitrophenyl-β-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 °C. At pH 5.0 and 40 °C the Km forpNPG was 0.1146 mM and it had a Vmax of 5.2792 nkatal.mg-1 protein (katal = 0.06 International Units = the amount of enzyme that produces, under standard conditions, one μmol 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-β-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-β-D-glucoside (a cyanogenic substrate), and sinigrin (a glucosinolate). The enzyme was not active against amygdalin, coniferin, or limonin glucoside.
Affiliations:
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Le document en format XML
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<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a">Purification and characterization of a beta-glucosidase from Citrus sinensis var. valencia fruit tissue</title>
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<author><name sortKey="Grohmann, Karel" sort="Grohmann, Karel" uniqKey="Grohmann K" first="Karel" last="Grohmann">Karel Grohmann</name>
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<series><title level="j" type="main">Journal of agricultural and food chemistry : (Print)</title>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>By product</term>
<term>Catalyst activity</term>
<term>Characterization</term>
<term>Chromatography, Gel</term>
<term>Citrus (enzymology)</term>
<term>Citrus sinensis</term>
<term>Enzymatic activity</term>
<term>Fruit</term>
<term>Hydrogen-Ion Concentration</term>
<term>Hydrolysis</term>
<term>Kinetics</term>
<term>Molecular Weight</term>
<term>Molecular mass</term>
<term>Optimum pH</term>
<term>Orange juice</term>
<term>Purification</term>
<term>Substrate specificity</term>
<term>Temperature</term>
<term>Tissue specificity</term>
<term>beta-Glucosidase (isolation & purification)</term>
<term>beta-Glucosidase (metabolism)</term>
<term>β-Glucosidase</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>
<keywords scheme="Pascal" xml:lang="fr"><term>Purification</term>
<term>Caractérisation</term>
<term>β-Glucosidase</term>
<term>Spécificité tissu</term>
<term>Fruit</term>
<term>Activité enzymatique</term>
<term>Spécificité substrat</term>
<term>Activité catalytique</term>
<term>pH optimum</term>
<term>Citrus sinensis</term>
<term>Masse moléculaire</term>
<term>Jus d'orange</term>
<term>Sous produit</term>
</keywords>
<keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Fruit</term>
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<front><div type="abstract" xml:lang="en">A preliminary survey demonstrated activity for α-D-glucosidase, α-D-mannosidase, α-L-arabinosidase, β-D-glucosidase, β-D-xylosidase, and β-D-galactosidase in orange fruit flavedo and albedo tissue, α-L-Rhamnosidase was not detected. Subsequently, a β-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 β-glucosidase exhibited low levels of activity against p-nitrophenyl-β-D-fucopyranoside (13.5%) and p-nitrophenyl-a-D-glucopyranoside (7.0%), compared to its activity against p-nitrophenyl-β-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 °C. At pH 5.0 and 40 °C the K<sub>m</sub>
forpNPG was 0.1146 mM and it had a V<sub>max</sub>
of 5.2792 nkatal.mg<sup>-1</sup>
protein (katal = 0.06 International Units = the amount of enzyme that produces, under standard conditions, one μmol 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-β-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-β-D-glucoside (a cyanogenic substrate), and sinigrin (a glucosinolate). The enzyme was not active against amygdalin, coniferin, or limonin glucoside.</div>
</front>
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<tree><country name="États-Unis"><noRegion><name sortKey="Cameron, Randall G" sort="Cameron, Randall G" uniqKey="Cameron R" first="Randall G." last="Cameron">Randall G. Cameron</name>
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<name sortKey="Baker, Robert A" sort="Baker, Robert A" uniqKey="Baker R" first="Robert A." last="Baker">Robert A. Baker</name>
<name sortKey="Grohmann, Karel" sort="Grohmann, Karel" uniqKey="Grohmann K" first="Karel" last="Grohmann">Karel Grohmann</name>
<name sortKey="Manthey, John A" sort="Manthey, John A" uniqKey="Manthey J" first="John A." last="Manthey">John A. Manthey</name>
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