Sequencing, cloning and high-yield expression of a fungal beta-N-acetylhexosaminidase in Pichia pastoris

0377436 - MBÚ 2013 RIV US eng J - Journal Article
Slámová, Kristýna - Bojarová, Pavla - Gerstorferová, Daniela - Fliedrová, Barbora - Hofmeisterová, Jana - Fiala, Martin - Pompach, Petr - Křen, Vladimír
Sequencing, cloning and high-yield expression of a fungal beta-N-acetylhexosaminidase in Pichia pastoris.
Protein Expression and Purification. Roč. 82, č. 1 (2012), s. 212-217. ISSN 1046-5928. E-ISSN 1096-0279
R&D Projects: GA ČR GP203/09/P024; GA ČR GD305/09/H008; GA ČR(CZ) GAP207/11/0629; GA MŠMT(CZ) 7E11011
Keywords : Talaromyces flavus * Pichia pastoris * Yeast expression system
Subject RIV: CE - Biochemistry
Impact factor: 1.429, year: 2012

The beta-N-acetylhexosaminidase from Talaromyces flavus has a remarkable synthetic ability, processing even carbohydrates with various functionalities. Its broader use is partially hampered by low-yield production in the native fungus. Here, we present an optimized 3-day production of this enzyme in the eukaryotic host of Pichia pastoris, in ca 10-fold higher volume activity (10 U/ml) and close-to-perfect purity (one chromatographic step needed). Importantly, the recombinant enzyme features the same biochemical and catalytic properties, including the syntheses with derivatized carbohydrate substrates. This is the first example of the overexpression of a fungal beta-N-acetylhexosaminidase by a single-cell producer in liquid medium. It represents a promising solution for wider biotechnological applications of this outstanding enzyme
Permanent Link: http://hdl.handle.net/11104/0209592