PpGal43A

Exo-Gal0056

(EC.3.2.1.145) exo-β-Galactosidase

CAZy Family: GH43

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~56 kDa)

Figure 1. Electrophoresis analysis of PpGal43A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coli BL21-pET28a-PpGal43A before IPTG induction; lane 2, culture lysate of E. coli BL21-pET28a-PpGal43A after IPTG induction; lane 3, PpGal43A  purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

0.72 U/mg protein (on pNP-β-Gal) at pH7.0 and 37°C.

One Unit of pNP-β-Gal activity is defined as the amount of enzyme required to release one μmole of p-nitrophenyl per minute from pNP-β-Gal (5 mM) in phosphate buffer (20 mM) pH 7.0.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of PpGal43A on different substrates.

Figure 2 TLC of PpGal43A hydrolyzed lactose. The concentration of lactose was 2 mg/mL in all reactions. The reactions were incubated in 20 mM sodium phosphate buffer (pH 7.0) for 12 hours.

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