BfGal36A (exo-α-Galactosidase)

BfGal36A

Ex-Gal0047

(EC.3.2.1.22)exo-α-Galactosidase

CAZy Family: GH36


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~82 kDa)

Figure 1. Electrophoresis analysis of BfGal36A. 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, BfGal36A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

1.86 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 sodium phosphate buffer (20 mM) pH7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfGal36A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100b±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_5

aReactions were performed with 5 mM substrate, pH 7.0, at 37 °C for10 min.

bAbsorption caused by released p-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:7.0

Temperature Optima:37°C


5.STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in sodiumphosphate (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.


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BfGal36A (exo-α-Galactosidase)

BfGal36A

Ex-Gal0047

(EC.3.2.1.22)exo-α-Galactosidase

CAZy Family: GH36


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~82 kDa)

Figure 1. Electrophoresis analysis of BfGal36A. 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, BfGal36A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

1.86 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 sodium phosphate buffer (20 mM) pH7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfGal36A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100b±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_5

aReactions were performed with 5 mM substrate, pH 7.0, at 37 °C for10 min.

bAbsorption caused by released p-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:7.0

Temperature Optima:37°C


5.STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in sodiumphosphate (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.