BfFuc29C (exo-α-Fucosidase )

BfFuc29C

Ex-Fuc0245-1

(EC 3.2.1.51) exo-α-Fucosidase

CAZy Family: GH29


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~48kDa)

Fig.1 Electrophoresis analysis ofBfFuc29C. 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, BfFuc29C purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

4.12 U/mg protein (on pNP-α-fuc) at pH 7.0 and 37°C.

One Unit of pNP-α-fuc activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-fuc (5 mM) in sodium phosphate buffer (20 mM) pH 7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfFuc29C on different substratesa.

Substrateb

Relative activity (%)c

pNPαGlc

_

pNPβGlc

_

pNPαGal

_

pNPβGal

_

pNPαMan

_

pNPβMan

_

pNPαXyl

_

pNPβXyl

_

pNPαAraf

_

pNPαArap

_

pNPαRha

_

pNPαFuc

100±0.0

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

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

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


4.PHYSICOCHEMICAL PROPERTIES

pH Optima:7.5

Temperature Optima:60°C


5.STORAGE CONDITIONS

The enzyme should be stored at-20°C. For assay, this enzyme should be diluted insodium phosphatebuffer (20 mM) pH7.0. Swirl to mix the enzyme immediately prior to use.


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BfFuc29C (exo-α-Fucosidase )

BfFuc29C

Ex-Fuc0245-1

(EC 3.2.1.51) exo-α-Fucosidase

CAZy Family: GH29


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~48kDa)

Fig.1 Electrophoresis analysis ofBfFuc29C. 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, BfFuc29C purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

4.12 U/mg protein (on pNP-α-fuc) at pH 7.0 and 37°C.

One Unit of pNP-α-fuc activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-fuc (5 mM) in sodium phosphate buffer (20 mM) pH 7.0.



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfFuc29C on different substratesa.

Substrateb

Relative activity (%)c

pNPαGlc

_

pNPβGlc

_

pNPαGal

_

pNPβGal

_

pNPαMan

_

pNPβMan

_

pNPαXyl

_

pNPβXyl

_

pNPαAraf

_

pNPαArap

_

pNPαRha

_

pNPαFuc

100±0.0

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

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

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


4.PHYSICOCHEMICAL PROPERTIES

pH Optima:7.5

Temperature Optima:60°C


5.STORAGE CONDITIONS

The enzyme should be stored at-20°C. For assay, this enzyme should be diluted insodium phosphatebuffer (20 mM) pH7.0. Swirl to mix the enzyme immediately prior to use.