BoXyl43A (exo-β-1,4-Xylosidase)

BoXyl43A

Ex-Xyl0164

(EC.3.2.1.37)exo-β-1,4-Xylosidase

CAZy Family: GH43


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~37 kDa)

Figure 1. Electrophoresis analysis of BoXyl43A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, BoXyl43A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

2.1 U/mg protein (on pNP-β-Xyl) at pH 7.0 and 37°C.

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



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES


Table 1. Relative activity ofBoXyl43A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

100±0.0

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

54±0.0

pNPαArap

_

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

bAbsorption caused by released p-nitrophenol was measured at 405nm. The relative activity on pNPαXyl was 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

Temperaturesuggestion: 37°C


5.STORAGE CONDITIONS

The enzyme should be stored at -20 °C. For assay, this enzyme should be diluted inphosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Zhou AD, Hu YB, et al. Characterization of a recombinant β-xylosidase of GH43 family from Bacteroides ovatus strain ATCC 8483[J]. Biocatalysis and Biotransformation, 2020, 38(1): 46-52.





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BoXyl43A (exo-β-1,4-Xylosidase)

BoXyl43A

Ex-Xyl0164

(EC.3.2.1.37)exo-β-1,4-Xylosidase

CAZy Family: GH43


PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~37 kDa)

Figure 1. Electrophoresis analysis of BoXyl43A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, BoXyl43A purified from Ni sepharose fastflow column.


2.SPECIFIC ACTIVITY

2.1 U/mg protein (on pNP-β-Xyl) at pH 7.0 and 37°C.

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



3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES


Table 1. Relative activity ofBoXyl43A on different substratesa.

Substrateb

Relative activity (%)c

pNPβGlu

_

pNPβGal

_

pNPβMan

_

pNPβXyl

100±0.0

pNPαGlc

_

pNPαGal

_

pNPαMan

_

pNPαAraf

54±0.0

pNPαArap

_

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

bAbsorption caused by released p-nitrophenol was measured at 405nm. The relative activity on pNPαXyl was 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

Temperaturesuggestion: 37°C


5.STORAGE CONDITIONS

The enzyme should be stored at -20 °C. For assay, this enzyme should be diluted inphosphate buffer (20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Zhou AD, Hu YB, et al. Characterization of a recombinant β-xylosidase of GH43 family from Bacteroides ovatus strain ATCC 8483[J]. Biocatalysis and Biotransformation, 2020, 38(1): 46-52.