Abstract:Various P-glycosidases, specific for the hydrolysis of isoflavone 7-O-p-glycosides, have been detected in the garbanzo bean, C i c e r a r i e t i n u m L. ,6-Monoglucosidases are present within the different plant organs in comparable activity whereas j3-diglycosidases are found in the leaves only. Both ,ô-glycosidase activities develop in the earliest States of plant growth, probably by protein de novo synthesis. A correlation between p-glycosidases and isofïavones present in garbanzo bean plants is discusse… Show more
“…The MALDI-TOF analysis also showed sequences of the five resultant peptide sequences are closest to those of β-glucosidase from C icer arietinum (Chickpea). The β-glucosidase from the C. arietinum has also been reported to exhibit the isoflavones conjugates hydrolyzing properties [24] confirming the functional nature of the enzyme purified from the Guar seeds.Fig. 6MALDI-TOF and MS analysis of purified enzyme ICHG showed maximum similarity with isoflavone-7-O-glucoside-specific beta-glucosidases from Cicer arietinum.…”
A β-glucosidase with high specific activity towards isoflavone glycosidic conjugates was purified from seeds of Guar (
Cyamopsis tetragonoloba
) by ammonium sulphate precipitation followed by size exclusion and ion exchange chromatography. The pH and temperature optima of the purified Isoflavones conjugate hydrolyzing β-glucosidase (ICHG) were found to be pH 4.5 and 37 °C, respectively. The enzyme was relatively stable at higher temperatures. Effect of different divalent metal ions was studied and it was found that Cobalt and Mercury ions completely inhibited the enzyme activity. K
m
and V
max
of the purified isoflavones conjugates hydrolyzing β-glucosidases (ICHG) was 0.86 mM and 6.6 IU/mg respectively. The enzyme was most likely a trimer (approximate Mr 150 kDa) with potential subunits of 50 kDa. The purified enzyme showed activity against isoflavone conjugate glycosides viz daidzin and genistin but was inactive towards other flavonoid conjugates. The product conversion was confirmed by HPTLC and HRMS analysis. The MALDI-TOF analysis of the ICHG showed a score greater than 78 with 20 matches in MASCOT software. The five resultant peptides obtained had highest similarity in sequence with β-glucosidase from C
icer arietinum
. The β-glucosidase from the
C. arietinum
has also been reported to exhibit the isoflavone conjugate hydrolyzing properties thus confirming the nature of the enzyme purified from the Guar seeds.
“…The MALDI-TOF analysis also showed sequences of the five resultant peptide sequences are closest to those of β-glucosidase from C icer arietinum (Chickpea). The β-glucosidase from the C. arietinum has also been reported to exhibit the isoflavones conjugates hydrolyzing properties [24] confirming the functional nature of the enzyme purified from the Guar seeds.Fig. 6MALDI-TOF and MS analysis of purified enzyme ICHG showed maximum similarity with isoflavone-7-O-glucoside-specific beta-glucosidases from Cicer arietinum.…”
A β-glucosidase with high specific activity towards isoflavone glycosidic conjugates was purified from seeds of Guar (
Cyamopsis tetragonoloba
) by ammonium sulphate precipitation followed by size exclusion and ion exchange chromatography. The pH and temperature optima of the purified Isoflavones conjugate hydrolyzing β-glucosidase (ICHG) were found to be pH 4.5 and 37 °C, respectively. The enzyme was relatively stable at higher temperatures. Effect of different divalent metal ions was studied and it was found that Cobalt and Mercury ions completely inhibited the enzyme activity. K
m
and V
max
of the purified isoflavones conjugates hydrolyzing β-glucosidases (ICHG) was 0.86 mM and 6.6 IU/mg respectively. The enzyme was most likely a trimer (approximate Mr 150 kDa) with potential subunits of 50 kDa. The purified enzyme showed activity against isoflavone conjugate glycosides viz daidzin and genistin but was inactive towards other flavonoid conjugates. The product conversion was confirmed by HPTLC and HRMS analysis. The MALDI-TOF analysis of the ICHG showed a score greater than 78 with 20 matches in MASCOT software. The five resultant peptides obtained had highest similarity in sequence with β-glucosidase from C
icer arietinum
. The β-glucosidase from the
C. arietinum
has also been reported to exhibit the isoflavone conjugate hydrolyzing properties thus confirming the nature of the enzyme purified from the Guar seeds.
“…The FGG activity parallels flavone accumulation. As described for the /3-glucosidase from Cicer arietinum (23), it decreases after having reached a maximum at stage IV. Kinetic studies revealed a significant preference for monoglucosides /3-linked at the 7-0-position of the flavonoid skeleton.…”
Section: The Addition Of Heavy Metal Ions Such Asmentioning
Flavone content and glucosidase activity were analyzed in various species of the genera Chamomilla, Matricaria, and Anthemis, especially during the development of the chamomile flower heads. The accumulation profile of flavonoids and the increase in enzyme activity were similar during ontogenesis. The accumulation of apigenin derivatives in closely related species was always linked to the occurrence of a catabolic beta-glucosidase in the respective plant organ. The flavone-glucoside-cleaving beta-glucosidase (FGG) from the ligulate florets of chamomile was purified to electrophoretic homogeneity by the following procedure: ammonium sulphate fractionation, anion exchange on Mono Q, hydrophobic interaction chromatography on Bio-Gel TSK Phenyl-5-PW, and gel filtration on Superose 12. The M (r) of the native enzyme was determined by gel filtration (500 kDa) and native PAGE (334 kDa). Only one subunit with an M (r) of 60 kDa could be detected after SDS-PAGE. The isoelectric point as determined by chromatofocussing on Mono P was at pH 4.6. During the purification procedure only one glucosidase activity appeared. A partially purified enzyme was used for characterization. The temperature optimum was at 37 degrees C and the pH-optimum 5.6. Energy of activation was 32.9 kJ/mol. The determination of the kinetic constants with various aryl glycosides proved a high affinity of the FGG towards flavone 7- O-glucosides. alpha-Glycosides and disaccharides were not hydrolyzed. Transglucosylation to an acceptor other than water was observed. Reagents interacting with sulfhydryl-groups strongly inhibited the enzyme.
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