SpMan92A(exo-α-1,2-Mannosidase)

SpMan92A

Ex-Man0092

(EC.3.2.1.113) exo-α-1,2-Mannosidase

CAZy Family: GH92


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~79 kDa)


Figure 1. Electrophoresis analysis ofSpMan92A. 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, SpMan92A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

2.71 U/mg protein (on pNP-α-Man) at pH 7.0 and 37°C

One Unit of pNP-α-Man activity is defined as the amount of enzyme required to release 1 nmol 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 SpMan92A on different substrates.

Substrate

Relative activity (%)

pNPβGlc

_

pNPβGal

-

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_



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



6. REFERENCES

[1]Robb M, Hobbs J K, Woodiga S A, et al. Molecular characterization of N-glycan degradation and transport in Streptococcus pneumoniae and its contribution to virulence[J]. PLoS pathogens, 2017, 13(1): e1006090.

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SpMan92A(exo-α-1,2-Mannosidase)

SpMan92A

Ex-Man0092

(EC.3.2.1.113) exo-α-1,2-Mannosidase

CAZy Family: GH92


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~79 kDa)


Figure 1. Electrophoresis analysis ofSpMan92A. 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, SpMan92A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

2.71 U/mg protein (on pNP-α-Man) at pH 7.0 and 37°C

One Unit of pNP-α-Man activity is defined as the amount of enzyme required to release 1 nmol 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 SpMan92A on different substrates.

Substrate

Relative activity (%)

pNPβGlc

_

pNPβGal

-

pNPβMan

_

pNPβXyl

_

pNPαGlc

_

pNPαGal

100±0.0

pNPαMan

_

pNPαAraf

_

pNPαArap

_



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



6. REFERENCES

[1]Robb M, Hobbs J K, Woodiga S A, et al. Molecular characterization of N-glycan degradation and transport in Streptococcus pneumoniae and its contribution to virulence[J]. PLoS pathogens, 2017, 13(1): e1006090.