BoAra51A (exo-α-Arabinofuranosidase)


BoAra51A

Ex-Ara0158

(EC.3.2.1) exo-α-Arabinofuranosidase

CAZy Family: GH51


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~57 kDa)

Figure 1. Electrophoresis analysis ofBoAra51A. 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, BoAra51A purified from Ni sepharose fastflow column.



2.SPECIFIC ACTIVITY

2.18 U/mg protein (on pNP-α-Araf) 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 BoAra51A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlc

17%

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

100±0

pNPαArap

_

aReactions were performed with 5mM substrate, pH 7.0, at 37°C for 5 min.

bAbsorption caused by releasedp-nitrophenol was measured at 405 nm. The relative activity on pNPαArawas taken as 100%.

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


4.PHYSICOCHEMICAL PROPERTIES

pH suggestion: 7.0

Temperature suggestion: 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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BoAra51A (exo-α-Arabinofuranosidase)


BoAra51A

Ex-Ara0158

(EC.3.2.1) exo-α-Arabinofuranosidase

CAZy Family: GH51


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~57 kDa)

Figure 1. Electrophoresis analysis ofBoAra51A. 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, BoAra51A purified from Ni sepharose fastflow column.



2.SPECIFIC ACTIVITY

2.18 U/mg protein (on pNP-α-Araf) 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 BoAra51A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlc

17%

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

100±0

pNPαArap

_

aReactions were performed with 5mM substrate, pH 7.0, at 37°C for 5 min.

bAbsorption caused by releasedp-nitrophenol was measured at 405 nm. The relative activity on pNPαArawas taken as 100%.

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


4.PHYSICOCHEMICAL PROPERTIES

pH suggestion: 7.0

Temperature suggestion: 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.