Crystal structure of a glycoside hydrolase family 68 β-fructosyltransferase from Beijerinckia indica subsp. indica in complex with fructose
| Autor: | Mika Nagaya, Keisuke Tamura, Takashi Tonozuka, Reika Kawai, Atsushi Nishikawa, Katsuaki Hirano, Tadashi Fujii, Junichi Kitamura, Takumi Tochio |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Stereochemistry
Crystal structure medicine.disease_cause Applied Microbiology and Biotechnology Biochemistry Analytical Chemistry law.invention 03 medical and health sciences chemistry.chemical_compound law medicine Glycoside hydrolase Crystallization Molecular Biology Escherichia coli 030304 developmental biology chemistry.chemical_classification Beijerinckia indica 0303 health sciences 030302 biochemistry & molecular biology Organic Chemistry Fructose General Medicine Enzyme chemistry Protein crystallization Biotechnology |
| DOI: | 10.6084/m9.figshare.12853038.v1 |
| Popis: | An enzyme belonging to glycoside hydrolase family 68 (GH68) from Beijerinckia indica subsp. indica NBRC 3744 was expressed in Escherichia coli. Biochemical characterization showed that the enzyme was identified to be a β-fructosyltransferase (BiBftA). Crystallization of a full-length BiBftA was initially attempted, but no crystals were obtained. We constructed a variant in which 5 residues (Pro199-Gly203) and 13 residues (Leu522-Gln534) in potentially flexible regions were deleted, and we successfully crystallized this variant BiBftA. BiBftA is composed of a five-bladed β-propeller fold as in other GH68 enzymes. The structure of BiBftA in complex with fructose unexpectedly indicated that one β-fructofuranose (β-Fruf) molecule and one β-fructopyranose molecule bind to the catalytic pocket. The orientation of β-Fruf at subsite −1 is tilted from the orientation observed in most GH68 enzymes, presenting a second structure of a GH68 enzyme in complex with the tilted binding mode of β-Fruf. Structure of the GH68 enzyme unexpectedly indicated that one β-fructofuranose and one β-fructopyranose bind to the catalytic pocket. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|