Cloning, Expression, Purification and Characterization of the β-galactosidase PoβGal35A from Penicillium oxalicum

• Published in: Molecular biotechnology Vol. 65; no. 7; pp. 1140 - 1150
• Main Authors: Zhou, Andong, Yi, Haoting, Yan, Xuecui, Mao, Zihan, Deng, Yannan, Lv, Xiao, Wang, Di, Zhang, Xin
• Format: Journal Article
• Language: English
• Published: New York Springer US 01.07.2023; Springer Nature B.V
• Subjects: Arabinogalactan; Biochemistry; Biological Techniques; Biotechnology; Calcium ions; Carbohydrates; Cell Biology; Chemistry; Chemistry and Materials Science; Cloning; Copper; Enzymes; Fungi; Galactose; Galactosidase; Glucose; Glycan; Glycerol; Glycoconjugates; Glycosidases; Glycoside hydrolase; Human Genetics; Original Paper; Penicillium oxalicum; Protein Science; Yeast; β-Galactosidase; β-galactosidase; GH35; Larchwood arabinogalactan; Galactan; Penicillium oxalicum
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-022-00620-y

Galactosidases are industrially important enzymes that hydrolyze galactosidic bonds in carbohydrates. Identifying new galactosidases with distinct functional characteristics is of paramount importance. In this study, we report the finding of a novel β-galactosidase PoβGal35A from the fungus Penicillium oxalicum . PoβGal35A belongs to the glycoside hydrolase family 35 (GH35), functions optimally at 70 °C and pH 5.0, and exhibits a specific high activity (191 ± 6.2 U/mg) towards p NPβgal. Ca 2+ , Fe 3+ and Ba 2+ ions enhance the activity of the enzyme, whereas Cu 2+ and Hg 2+ significantly reduce it. This enzyme releases galactose from β-1,3-galactan, β-1,4-galactan, β-1,6-galactan, as well as arabinogalactan from larchwood (LWAG). In addition, PoβGal35A acts synergistically with arabinosidase to degrade LWAG. These results suggest that PoβGal35A is a high activity exo-β-1,3/4/6-galactanase that can be used to establish glycan blocks in glycoconjugates, and thus provides a new tool for biotechnological applications.