PpXyl31A

Ex-Gal0162

(EC.3.2.1)exo-α-1,6-Xylosidase

CAZy Family: GH31

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~101 kDa)

Figure 1. Electrophoresis analysis of PpXyl31A. 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, PpXyl31A purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

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

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

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity ofPpXyl31A on different substrates^a.

^aReactions were performed with 5mM 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 pNPαXyl was taken as 100%.

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

4.PHYSICOCHEMICAL PROPERTIES

pH Optima:7.0

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) pH 7.0. Swirl to mix the enzyme immediately prior to use.