PpRha
Exo-Rha0247
(EC.3.2.1.40) exo-α-Rhamnosidase
CAZy Family: GH78
PROPERTIES
1. ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~104 kDa)
Figure 1. Electrophoresis analysis of PpRha. 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, PpRha purified from Ni sepharose fastflow column.
2. SPECIFIC ACTIVITY
1.31 U/mg protein (on pNP-α-Rha) at pH 7.0 and 37°C
One Unit of pNP-α-Rha activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-Rha (5 mM) in phosphate buffer (20 mM) pH 7.0.
3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of ppRha on different substrates.
Substrate |
Relative activity (%) |
pNPβGlu |
_ |
pNPβGal |
_ |
pNPβMan |
_ |
pNPβXyl |
_ |
pNPαGlu |
_ |
pNPαGal |
_ |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
_ |
pNPαRha |
100.0±0.6 |
4. PHYSICOCHEMICAL PROPERTIES
pH Optima: suggested use 7.0
Temperature Optima: suggested use 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.
PpRha
Exo-Rha0247
(EC.3.2.1.40) exo-α-Rhamnosidase
CAZy Family: GH78
PROPERTIES
1. ELECTROPHORETIC PURITY
-Single band on SDS-gel electrophoresis (MW ~104 kDa)
Figure 1. Electrophoresis analysis of PpRha. 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, PpRha purified from Ni sepharose fastflow column.
2. SPECIFIC ACTIVITY
1.31 U/mg protein (on pNP-α-Rha) at pH 7.0 and 37°C
One Unit of pNP-α-Rha activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-Rha (5 mM) in phosphate buffer (20 mM) pH 7.0.
3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES
Table 1. Relative activity of ppRha on different substrates.
Substrate |
Relative activity (%) |
pNPβGlu |
_ |
pNPβGal |
_ |
pNPβMan |
_ |
pNPβXyl |
_ |
pNPαGlu |
_ |
pNPαGal |
_ |
pNPαMan |
_ |
pNPαAraf |
_ |
pNPαArap |
_ |
pNPαRha |
100.0±0.6 |
4. PHYSICOCHEMICAL PROPERTIES
pH Optima: suggested use 7.0
Temperature Optima: suggested use 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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