3172 J. Agric. Food Chem., Vol. 51, No. 10, 2003
Odoux et al.
LITERATURE CITED
Table 3. Effect of Metal Ions on Vanilla Bean â-D-Glucosidase
(
1) Arana, F. E. Action of a â-glucosidase in the curing of vanilla.
concn
tested
(mM)
concn
tested
(mM)
Food Res. 1943, 8, 343-351.
relative
activity
relative
activitya
a
(2) Ranadive, A. S.; Szkutnica, K.; Guerrera, J. G.; Frenkel, C.
Vanillin biosynthesis in vanilla beans. In Proceedings of the 9th
International Congress on Essential Oils, Singapore, March 13-
17, 1983; Essential Oils Association of Singapore: Singapore,
1983; Book 2, pp 147-154.
(3) Odoux, E. Changes in vanillin and glucovanillin concentrations
during the various stages of the process traditionally used for
curing Vanilla fragrans beans in R e´ union. Fruits 2000, 55, 119-
ion
ion
Hg2+
0.001
78
58
29
6
3
112
29
Mg
Ca
2+
1
1
1
1
1
1
1
89
82
47
34
30
7
2+
0
0
0
0
.005
.02
.04
.06
2
+
Co
Mn
Ni
2+
2+
Ag+
0.1
1
Zn
2+
2+
Cu
4
1
25.
a
The â-glucosidase (0.05 nkat in 200 µL of 50 mM HEPES−NaOH buffer, pH
(
(
(
4) Leong, G.; Archavlis, A.; Derbesy, M. Research on the glucoside
fraction of the vanilla bean. J. Essent. Oil Res. 1989, 1, 33-41.
5) Kanisawa, T. Flavor development in vanilla beans. Kouryou
7
.0) was exposed for 30 min at 4 °C to different concentrations of metal ions. The
activity was then measured under standard conditions and expressed relative to
the activity measured on p-nitrophenyl-â-D-glucopyranoside (100) without added
ion.
1
993, 180, 113-123.
6) Wild-Altamiro, C. Enzymic activity during growth of vanilla
fruit. 1. Proteinase, glucosidase, peroxidase and polyphenoloxi-
dase. J. Food Sci. 1969, 34, 235-238.
7) Hanum, T. Changes in vanillin and activity of â-glucosidase and
oxidases during post harvest processing of vanilla beans (Vanilla
planifolia). Bull. Teknol. Ind. Pangan 1997, 8, 46-52.
8) Dignum, M. J. W.; Kerler, J.; Verpoorte, R. â-glucosidase and
peroxidase stability in crude enzyme extracts from green beans
of Vanilla planifolia Andrews. Phytochem. Anal. 2001, 12, 174-
179.
Table 4. Relative Activity of Bean â-D-Glucosidase on Various
Substrates
(
relative
activitya
substrate
(
p-nitrophenyl-â-D-glucopyranoside
p-nitrophenyl-â-D-fucopyranoside
p-nitrophenyl-â-D-galactopyranoside
p-nitrophenyl-â-D-xylopyranoside
p-nitrophenyl-â-D-mannopyranoside
p-nitrophenyl-R-L-arabinofuranoside
p-nitrophenyl-R-L-rhamnopyranoside
p-nitrophenyl-R-D-glucopyranoside
100
215
61
5
b
nd
(
9) Lowry, O. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J.
Protein measurement with the Folin-phenol reagent. J. Biol.
Chem. 1951, 193, 265-275.
nd
nd
nd
22
300
92
c
glucovanillin
(10) Laemmli, U. K. Cleavage of structural proteins during the
c
prunasin
assembly of the head of bacteriophage T4. Nature 1970, 227,
c
esculin
6
80-685.
c,d
salicin
3.6
nd
c,d
(11) Guivarc’h, A.; Caissard, J. C.; Azmi, A.; Elmayan, T.; Chriqui,
D.; Tepfer, M. In situ detection of expression of the gus reporter
gene in transgenic plants: ten years of blue genes. Transgenic
Res. 1996, 5, 281-288.
phloridzin
c,d
indoxyl-â-D-glucopyranoside
nd
c-e
quercetin-3-â-D-glucopyranoside
nd
d,f
amygdalin
nd
d,f
octyl-â-D-glucopyranoside
0.4
nd
d,f
(12) Sigma-Aldrich, Inc. Glucose (GO) assay kit; Technical Bulletin,
cellobiose
d,f
2002.
gentiobiose
nd
(
13) Cameron, R. G.; Manthey, J. A.; Baker, R. A.; Grohmann, K.
Purification and characterization of a beta-glucosidase from
Citrus sinensis var. Valencia fruit tissue. J. Agric. Food Chem.
2001, 49, 4457-4462.
a
2
mM substrate concentration, 0.1 M sodium phosphate buffer (pH 7.0), 20
min, 40 °C, 0.1 nkat of enzyme. Activity expressed relative to activity measured
b
c
on p-nitrophenyl-â-D-glucopyranoside (100). Not detected. Released aglycon
d
e
measured by HPLC. Incubated with the enzyme for 5 h. Assayed in the presence
of 10% ethanol. Released glucose measured by the glucose oxidase/peroxidase/
(14) Konno, H.; Yamasaki, Y.; Katoh, K. A â-glucosidase associated
with cellwalls from cell suspension cultures of carrot. Phy-
tochemistry 1996, 43, 1157-1161.
f
o-dianisidine system.
(
15) Gus-Mayer, S.; Brunner, H.; Schneider-Poetsch, H. A. W.;
R u¨ diger, W. Avenacosidase from oat: purification, sequence
analysis and biochemical characterization of a new member of
the BGA family of â-glucosidases. Plant Mol. Biol. 1994, 26,
909-921.
from butter bean, Phaseolus lunatus (21), and wheat seedlings
(17). Broad specificity of plant â-glucosidases with regard to
p-nitrophenyl glycosides was also observed in sweet cherry (25),
maize (26), and Polygonum tinctorium leaves (27) with maxi-
mum activity found on p-nitrophenyl-â-D-fucopyranoside. No
activity was found on p-nitrophenyl-â-D-mannopyranoside, R-L-
arabinofuranoside, R-L-rhamnopyranoside, and R-D-glucopyra-
noside. Like rubber tree (20) and maize (26) â-glucosidases,
the vanilla bean â-glucosidase does not hydrolyze glycosyl
glucosides, cellobiose, gentiobiose, and amygdalin (D-mande-
lonitrile-â-gentiobioside). Thus, according to its specificity, this
vanilla enzyme can be viewed more as a â-D-glycosidase than
as a â-D-glucosidase. Rates of hydrolysis of prunasin, esculin,
glucovanillin, and salicin at 2 mM were 3.00:0.92:0.22:0.04
relative to pNPG, which was unexpected since the aglycon of
salicin is structurally closer to vanillin than esculin. Similar
specificity was observed in the case of wheat seedling â-glu-
cosidase (17). Phloridzin, indoxyl-â-D-glucopyranoside, and
quercetin-3-â-D-glucopyranoside were not substrates. Octyl-â-
D-glucopyranoside was hydrolyzed at a very slow rate (0.4%
vs pNPG).
(16) H o¨ sel, W.; Tober, I.; Eklund, S. H.; Conn, E. E. Characterization
of b-glucosidases with high specificity for the cyanogenic
glucoside dhurrin in Sorghum bicolor (L.) Moench seedlings.
Arch. Biochem. Biophys. 1987, 252, 152-162.
(
(
(
(
17) Sue, M.; Ishihara, A.; Iwamura, H. Purification and characteriza-
tion of a hydroxamic acid glucoside â-glucosidase from wheat
(Triticum aestiVum L.) seedlings. Planta 2000, 210, 432-438.
18) Dubois, M.; Gilles, K. A.; Hamilton, J. K.; Rebers, P. A.; Smith,
F. Colorimetric method for determination of sugars and related
substances. Anal. Chem. 1956, 28, 350-356.
19) Ranjoanizafy, X. Purification et e´ tude de quelques propri e´ t e´ s
de la â-glucosidase de la Vanille. Implications dans la pr e´ para-
tion de la Vanille; ENSIA-SIARC: Montpellier, France. 1997.
20) Selmar, D.; Lieberei R.; Biehl, B.; Voigt, J. HeVea linamarases
A nonspecific â-glycosidase. Plant Physiol. 1987, 83, 557-563.
(21) Itoh-Nashida, T.; Hiraiwa, M.; Uda, Y. Purification and properties
of (linamarase) from the butter bean, Phaseolus lunatus. J.
Biochem. 1987, 101, 847-854.