4TMB

CRYSTAL STRUCTURE of OLD YELLOW ENZYME from CANDIDA MACEDONIENSIS AKU4588

4TMB の概要

• エントリーDOI: 10.2210/pdb4tmb/pdb
• 関連するPDBエントリー: 4tmc
• 分子名称: Old yellow enzyme, FLAVIN MONONUCLEOTIDE (3 entities in total)
• 機能のキーワード: tim barrel motif, dehydrogenase, flavoprotein
• 由来する生物種: Kluyveromyces marxianus (Yeast)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 185615.96

構造登録者

Horita, S.,Kataoka, M.,Kitamura, N.,Nakagawa, T.,Miyakawa, T.,Ohtsuka, J.,Nagata, K.,Shimizu, S.,Tanokura, M. (登録日: 2014-05-31, 公開日: 2015-02-11, 最終更新日: 2023-11-08)

主引用文献

Horita, S.,Kataoka, M.,Kitamura, N.,Nakagawa, T.,Miyakawa, T.,Ohtsuka, J.,Nagata, K.,Shimizu, S.,Tanokura, M.
An Engineered Old Yellow Enzyme that Enables Efficient Synthesis of (4R,6R)-Actinol in a One-Pot Reduction System
Chembiochem, 16:440-445, 2015

PubMed Abstract: (4R,6R)-Actinol can be stereo-selectively synthesized from ketoisophorone by a two-step conversion using a mixture of two enzymes: Candida macedoniensis old yellow enzyme (CmOYE) and Corynebacterium aquaticum (6R)-levodione reductase. However, (4S)-phorenol, an intermediate, accumulates because of the limited substrate range of CmOYE. To address this issue, we solved crystal structures of CmOYE in the presence and absence of a substrate analogue p-HBA, and introduced point mutations into the substrate-recognition loop. The most effective mutant (P295G) showed two- and 12-fold higher catalytic activities toward ketoisophorone and (4S)-phorenol, respectively, than the wild-type, and improved the yield of the two-step conversion from 67.2 to 90.1%. Our results demonstrate that the substrate range of an enzyme can be changed by introducing mutation(s) into a substrate-recognition loop. This method can be applied to the development of other favorable OYEs with different substrate preferences.

PubMed: 25639703
DOI: 10.1002/cbic.201402555

実験手法

X-RAY DIFFRACTION (1.8 Å)