BtGalEPI

Endo-Gal0254

(EC:3.2.1)endo-α-1,6-Galactosidase

CAZy Family:EPI

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~42 kDa)

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

2.SPECIFIC ACTIVITY

0.001 U/mg protein (on α-1,6-galactan) at pH 7.0and 37°C.

One Unit of α-1,6-galactanase activity is defined as the amount of enzyme required to release 1 μmol of reducing sugars per minute from α-1,6-galactan (1 mg/ml) in PBS buffer pH 7.0.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

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

^aReactions were performed with 5 mM (p-nitrophenyl glycosides) or 1 mg/ml (polysaccharides), pH 7.0, at 37°C for 10 min.

^bAbsorption caused by released p-nitrophenol or reducing sugars was measured at 405 or 540 nm. The relative activity on α-1,6-galactan was taken as 100%.

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 37°C

5.STORAGE CONDITIONS

The enzyme is supplied in PBS buffer and should be stored at -20°C. For assay, this enzyme should be diluted in PBS buffer pH 7.0. Swirl to mix the enzyme immediately prior to use.