BtGal36B (exo-α-Galactosidase)


BtGal36B

Ex-Gal0038

(EC.3.2.1.22)exo-α-Galactosidase

CAZy Family:GH36


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~74kDa)

Figure 1. Electrophoresis analysis ofBtGal36B. 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,BtGal36Bpurified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

0.06 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) pH7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity ofBtGal36Bon different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_

aReactions were performed with 5 mM substrate, pH 7.0, at 30°Cfor 30 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: 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 (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

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BtGal36B (exo-α-Galactosidase)


BtGal36B

Ex-Gal0038

(EC.3.2.1.22)exo-α-Galactosidase

CAZy Family:GH36


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~74kDa)

Figure 1. Electrophoresis analysis ofBtGal36B. 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,BtGal36Bpurified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

0.06 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) pH7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity ofBtGal36Bon different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_

aReactions were performed with 5 mM substrate, pH 7.0, at 30°Cfor 30 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: 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 (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.