BfGal27A
Ex-Gal0046
(EC.3.2.1.22)exo-α-Galactosidase
CAZy Family: GH27
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~56kDa)
Figure 1. Electrophoresis analysis of BfGal27A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, BfGal27A purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
2.66 U/mg protein (on pNP-α-gal) at pH 7.0 and 37°C
One Unit of pNP-α-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-gal (5 mM) in phosphate buffer (20 mM) pH 7.0.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of BfGal27Aon different substratesa.
Substrateb |
Relative activity (%)c |
pNPβGlu |
_ |
pNPβGal |
_ |
pNPβMan |
_ |
pNPβXyl |
_ |
pNPαGlc |
_ |
pNPαGal |
100±0 |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
_ |
aReactions were performed with 5mM substrate, pH7.0, at 37°C for 5 min.
bAbsorption caused by releasedp-nitrophenol was measured at 405 nm. The relative activity on pNPGal was taken as 100%.
cThe data are reported as means±standard errors from the mean for three independent experiments.
4.PHYSICOCHEMICAL PROPERTIES
pH Optima: suggesteduse 7.0
Temperature Optima: suggesteduse 37°C
5.STORAGE CONDITIONS
The enzyme should be stored at -20 °C. For assay, this enzyme should be diluted in phosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.
BfGal27A
Ex-Gal0046
(EC.3.2.1.22)exo-α-Galactosidase
CAZy Family: GH27
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~56kDa)
Figure 1. Electrophoresis analysis of BfGal27A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, BfGal27A purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
2.66 U/mg protein (on pNP-α-gal) at pH 7.0 and 37°C
One Unit of pNP-α-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-gal (5 mM) in phosphate buffer (20 mM) pH 7.0.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of BfGal27Aon different substratesa.
Substrateb |
Relative activity (%)c |
pNPβGlu |
_ |
pNPβGal |
_ |
pNPβMan |
_ |
pNPβXyl |
_ |
pNPαGlc |
_ |
pNPαGal |
100±0 |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
_ |
aReactions were performed with 5mM substrate, pH7.0, at 37°C for 5 min.
bAbsorption caused by releasedp-nitrophenol was measured at 405 nm. The relative activity on pNPGal was taken as 100%.
cThe data are reported as means±standard errors from the mean for three independent experiments.
4.PHYSICOCHEMICAL PROPERTIES
pH Optima: suggesteduse 7.0
Temperature Optima: suggesteduse 37°C
5.STORAGE CONDITIONS
The enzyme should be stored at -20 °C. For assay, this enzyme should be diluted in phosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.
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