PoGal43A (PoGal3)

Endo-Gal0054

(EC. 3.2.1.145) exo-β-1,3-Galactanase

CAZy Family: GH43

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~49 kDa)

Figure 1. Electrophoresis analysis of PoGal43A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before IPTG induction; lane 2-4, culture lysat after IPTG induction; lane 5, PoGal43A purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

0.09 U/mg protein (on AG-P-I) at pH 5.0 and 37°C.

One Unit of galactanase activity is defined as the amount of enzyme required to release 1 μmol of reducing sugar per minute from AG-P-I (5 mg/ml) in Na-Acetate buffer (20 mM) pH 5.0.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1. Substrate specificity of recombinant PoGal3 towards different polysaccharides.

Figure2. HPAEC of degradation products of different polysaccharides hydrolyzed by PoGal3. Reactions were incubated with recombinant enzyme at 40°C for 12 h. The bottom of each graph was the standard sample of galactose or 1,6-galactoise, and oligosaccharides produced in the reaction were detected by HPAEC at 0 h and 12 h.

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 5.0

pH Stability: 2.0-11.0

Temperature Optima: 45°C

5. STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in acetate buffer (20 mM) pH 4.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Zhou T, Hu YB, Zhou YF, et al. Molecular Cloning and Characterization of a Novel Exo-β-1,3-Galactanase from Penicillium oxalicum sp. 68 J. Microbiol. Biotechnol. 2022. 32(8): 1064–1071.