|
|||||
|
|
Structural basis of different substrate preferences of two old yellow enzymes from yeasts in the asymmetric reduction of enone compounds |
|
| Authors: | Shoichiro Horita Michihiko Kataoka Nahoko Kitamura Takuya Miyakawa Jun Ohtsuka Yuko Maejima |
| Affiliation: | 1. Department of Applied Biologic al Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan;2. Department of Bioregulation and Pharmacological Medicine, Fukushima Medical University School of Medicine, Fukushima, Japan;3. Division of Applied Life Sciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan;4. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, Japan;5. Department of Bioregulation and Pharmacological Medicine, Fukushima Medical University School of Medicine, Fukushima, Japan |
| Abstract: | Old yellow enzymes (OYEs) are potential targets of protein engineering for useful biocatalysts because of their excellent asymmetric reductions of enone compounds. Two OYEs from different yeast strains, Candida macedoniensis AKU4588 OYE (CmOYE) and Pichia sp. AKU4542 OYE (PsOYE), have a sequence identity of 46%, but show different substrate preferences; PsOYE shows 3.4-fold and 39-fold higher catalytic activities than CmOYE toward ketoisophorone and (4S)-phorenol, respectively. To gain insights into structural basis of their different substrate preferences, we have solved a crystal structure of PsOYE, and compared its catalytic site structure with that of CmOYE, revealing the catalytic pocket of PsOYE is wider than that of CmOYE due to different positions of Phe246 (PsOYE)/Phe250 (CmOYE) in static Loop 5. This study shows a significance of 3D structural information to explain the different substrate preferences of yeast OYEs which cannot be understood from their amino acid sequences. Abbreviations: OYE: Old yellow enzymes, CmOYE: Candida macedoniensis AKU4588 OYE, PsOYE: Pichia sp. AKU4542 OYE |
| Keywords: | Biocatalysis enzyme catalysis enzymes old yellow enzyme X-ray crystallography |