BtGal27D

Ex-Gal0035

(EC.3.2.1.22) exo-α-Galactosidase

CAZy Family: GH27

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~92kDa)

Figure 1. Electrophoresis analysis of BtGal27D. 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,BtGal27D purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

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

One Unit of pNP-α-gal activity is defined as the amount of enzyme required to release 1 μmol of p-nitrophenyl per minute from pNP-α-gal (5 mM) insodium phosphate buffer(20 mM) pH7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtGal27D on different substrates^a.

^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 405nm. The relative activity on pNPαGal 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 insodium phosphatebuffer(20 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.