BlGal43A

Ex-Gal0057

(EC. 3.2.1.145) exo-β-1,3-Galactosidase

CAZy Family: GH43

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~100 kDa)

Figure 1. Electrophoresis analysis of BlGal43A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate of E. coli BL21-pET28a-BlGal43A before IPTG induction; lane 2, culture lysate of E. coli BL21-pET28a-BlGal43A after IPTG induction; lane 3, BlGal43A purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

16.0 U/mg protein (on LWAG) at ph5.5 and 40°C.

One unit of enzyme activity toward polysaccharides was defined as the amount of enzyme required to produce 1 μmol of reducing sugars per minute.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Relative activity of BtGal43B on different substrates.

Figure 2 TLC of BlGal43A hydrolyzed lactose. The concentration of lactose was 2 mg/mL in all reactions. The reactions were incubated in 20 mM sodium phosphate buffer (pH 5.5) for 12 hours.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: suggested use 5.5

Temperature Optima: suggested use 40°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 5.5. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Fujita, Kiyotaka, et al. Bifidobacterium longum subsp. longum exo-β-1, 3-galactanase, an enzyme for the degradation of type II arabinogalactan. Applied and Environmental Microbiology 80.15 (2014): 4577-4584.