BtGal92B
endo-Gal0262
(EC.3.2.1)endo-α-1,6-galactanase
CAZy Family: GH92
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~85 kDa)
Figure 1. Electrophoresis analysis of BtGal92B. 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, BtGal92B purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
0.001 U/mg protein (on α-1,6-galactan) at pH 7.0 and 37°C.
One Unit of α-1,6-galactanase activity is defined as the amount of enzyme required to release 1 μmol of galactose per minute from α-1,6-galactan (1 mg/ml) in PBS buffer pH 7.0.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of BtGal92B on different substratesa.
Substrate |
Relative activity(%) |
pNPβGlc |
- |
pNPβGal |
- |
pNPβMan |
_ |
pNPβXyl |
- |
pNPαGlc |
_ |
pNPαGal |
_ |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
- |
pNPαRha |
- |
pNPαFuc |
- |
α-1,6-galactan |
100±0.0 |
β-1,4-galactan |
- |
α-1.4-galacturonic |
- |
α-1.4-glucan |
- |
β-1.3-glucan |
- |
β-1.6-glucan |
- |
aReactions were performed with5 mM (p-nitrophenyl glycosides) or 1 mg/ml (polysaccharides), pH 7.0, at 37°C for 10 min.
bAbsorption caused by released p-nitrophenol or reducing sugars was measured at 405 or 540 nm. The relative activity on α-1,6-galactan) was taken as 100%.
4.PHYSICOCHEMICAL PROPERTIES
pH Optima: 7.0
Temperature Optima: 37°C
5.STORAGE CONDITIONS
The enzyme is supplied in PBS buffer and should be stored at -20°C. For assay, this enzyme should be diluted in PBS buffer pH 7.0. Swirl to mix the enzyme immediately prior to use.
BtGal92B
endo-Gal0262
(EC.3.2.1)endo-α-1,6-galactanase
CAZy Family: GH92
PROPERTIES
1.ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~85 kDa)
Figure 1. Electrophoresis analysis of BtGal92B. 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, BtGal92B purified from Ni sepharose fastflow column.
2.SPECIFIC ACTIVITY
0.001 U/mg protein (on α-1,6-galactan) at pH 7.0 and 37°C.
One Unit of α-1,6-galactanase activity is defined as the amount of enzyme required to release 1 μmol of galactose per minute from α-1,6-galactan (1 mg/ml) in PBS buffer pH 7.0.
3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of BtGal92B on different substratesa.
Substrate |
Relative activity(%) |
pNPβGlc |
- |
pNPβGal |
- |
pNPβMan |
_ |
pNPβXyl |
- |
pNPαGlc |
_ |
pNPαGal |
_ |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
- |
pNPαRha |
- |
pNPαFuc |
- |
α-1,6-galactan |
100±0.0 |
β-1,4-galactan |
- |
α-1.4-galacturonic |
- |
α-1.4-glucan |
- |
β-1.3-glucan |
- |
β-1.6-glucan |
- |
aReactions were performed with5 mM (p-nitrophenyl glycosides) or 1 mg/ml (polysaccharides), pH 7.0, at 37°C for 10 min.
bAbsorption caused by released p-nitrophenol or reducing sugars was measured at 405 or 540 nm. The relative activity on α-1,6-galactan) was taken as 100%.
4.PHYSICOCHEMICAL PROPERTIES
pH Optima: 7.0
Temperature Optima: 37°C
5.STORAGE CONDITIONS
The enzyme is supplied in PBS buffer and should be stored at -20°C. For assay, this enzyme should be diluted in PBS buffer pH 7.0. Swirl to mix the enzyme immediately prior to use.
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