PoGal30A (PoGal30)

Endo-Gal0066

(EC. 3.2.1.164) endo-β-1,6-Galactanase

CAZy Family: GH30

PROPERTIES

1. ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~53 kDa)

Figure 1. Electrophoresis analysis of PoGal30A. 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, PoGal30A purified from Ni sepharose fastflow column.

2. SPECIFIC ACTIVITY

0.54 U/mg protein (on dGA) at pH 4.0 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 dGA (2 mg/ml) in Na-Acetate buffer (20 mM) pH 5.0.

3. RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Figure 2. HPAEC-PAD analysis of hydrolysis products of galactobiose generated by PoGal30A

Figure 3. HPAEC-PAD analysis of the products of the degradation of Galactan by PoGal30A

4. PHYSICOCHEMICAL PROPERTIES

pH Optima: 4.0

pH Stability: 3.0-6.0

Temperature Optima: 40°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] Zhang X, Yuan Y, et al. Cloning, Expression, and Characterization of Endo‑β‑1,6‑galactanase PoGal30 from Penicillium oxalicum. Applied Biochemistry and Biotechnology. 2022.