Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
5914653 | Journal of Structural Biology | 2011 | 8 Pages |
Abstract
We have determined the crystal structure of Trichoderma reesei (Hypocrea jecorina) β-galactosidase (Tr-β-gal) at a 1.2 Ã
resolution and its complex structures with galactose, IPTG and PETG at 1.5, 1.75 and 1.4Â Ã
resolutions, respectively. Tr-β-gal is a potential enzyme for lactose hydrolysis in the dairy industry and belongs to family 35 of the glycoside hydrolases (GH-35). The high resolution crystal structures of this six-domain enzyme revealed interesting features about the structure of Tr-β-gal. We discovered conformational changes in the two loop regions in the active site, implicating a conformational selection-mechanism for the enzyme. In addition, the Glu200, an acid/base catalyst showed two different conformations which undoubtedly affect the pKa value of this residue and the catalytic mechanism. The electron density showed extensive glycosylation, suggesting a structure stabilizing role for glycans. The longest glycan showed an electron density that extends to the eighth monosaccharide unit in the extended chain. The Tr-β-gal structure also showed a well-ordered structure for a unique octaserine motif on the surface loop of the fifth domain.
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Authors
Mirko Maksimainen, Nina Hakulinen, Johanna M. Kallio, Tommi Timoharju, Ossi Turunen, Juha Rouvinen,