PpRha(exo-α-Rhamnosidase)

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)

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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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PpRha(exo-α-Rhamnosidase)

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)

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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.