BvAra51A (exo-α-Arabinofuranosidase)

BvAra51A

Ex-Ara0154

(EC.3.2.1.145)exo-α-Arabinofuranosidase

CAZy Family: GH51


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~71kDa)

Figure 1. Electrophoresis analysis of BvAra51A. 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, BvAra51Apurified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

1.73 U/mg protein (on pNP-α-Ara) at pH 7.0 and 37oC.

One Unit of pNP-α-Araactivity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-Ara(5 mM) in phosphate buffer(20 mM) pH 7. 0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BvAra51A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

_

pNPαArap

100±0

aReactions were performed with 5 mM substrate, pH7.0, at 37°C for 5 min.

bAbsorption caused by released p-nitrophenol was measured at 405nm. The relative activity on pNPAra was taken as 100%.

cThe data are reported as means±standard errors from the mean for three independent experiments.


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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BvAra51A (exo-α-Arabinofuranosidase)

BvAra51A

Ex-Ara0154

(EC.3.2.1.145)exo-α-Arabinofuranosidase

CAZy Family: GH51


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~71kDa)

Figure 1. Electrophoresis analysis of BvAra51A. 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, BvAra51Apurified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

1.73 U/mg protein (on pNP-α-Ara) at pH 7.0 and 37oC.

One Unit of pNP-α-Araactivity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-Ara(5 mM) in phosphate buffer(20 mM) pH 7. 0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BvAra51A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

_

pNPαArap

100±0

aReactions were performed with 5 mM substrate, pH7.0, at 37°C for 5 min.

bAbsorption caused by released p-nitrophenol was measured at 405nm. The relative activity on pNPAra was taken as 100%.

cThe data are reported as means±standard errors from the mean for three independent experiments.


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.