BfMan92A (exo-α-1,2-Mannosidase)


BfMan92A

Ex-Man0085

(EC.3.2.1.113)exo-α-1,2-Mannosidase

CAZy Family: GH92


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~86 kDa)


Figure 1. Electrophoresis analysis of BfMan92A. 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, BfMan92A. purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

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

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



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfMan92A. on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

100%±0.0

pNPαAraf

_

pNPαArap

_

aReactions were performed with 5 mM substrate, pH 7.0, at 37°C for 10 min.

bAbsorption caused by released p-nitrophenol was measured at 405 nm.

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


4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 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) pH7.0. Swirl to mix the enzyme immediately prior to use.


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BfMan92A (exo-α-1,2-Mannosidase)


BfMan92A

Ex-Man0085

(EC.3.2.1.113)exo-α-1,2-Mannosidase

CAZy Family: GH92


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~86 kDa)


Figure 1. Electrophoresis analysis of BfMan92A. 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, BfMan92A. purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

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

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



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BfMan92A. on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

_

pNPαMan

100%±0.0

pNPαAraf

_

pNPαArap

_

aReactions were performed with 5 mM substrate, pH 7.0, at 37°C for 10 min.

bAbsorption caused by released p-nitrophenol was measured at 405 nm.

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


4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 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) pH7.0. Swirl to mix the enzyme immediately prior to use.