Benign synthesis of 2-ethylhexanoic acid by cytochrome P450cam: Enzymatic, crystallographic, and theoretical studies
Benign synthesis of 2-ethylhexanoic acid by cytochrome P450cam: Enzymatic, crystallographic, and theoretical studies
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DOI：
10.1021/bi010063
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发表时间：
2001-08-14
期刊：
影响因子：
2.9
通讯作者：
Jones, JP
中科院分区：
文献类型：
--
作者：
French, KJ;Strickler, MD;Jones, JP
This study examines the ability of P450cam. to catalyze the formation of 2-ethylhexanoic acid from 2-ethylhexanol relative to its activity on the natural substrate camphor. As is the case for camphor, the P450cam exhibits stereoselectivity for binding (R)- and (S)-2-ethylhexanol. Kinetic studies indicate (R)-2-ethylhexanoic acid is produced 3.5 times as fast as the (S)-enantiomer. In a racemic mixture of 2-ethylhexanol, P450cam produces 50% more (R)-2-ethylhexanoic acid than (S)-2-ethylhexanoic acid. The reason for stereoselective 2-ethylhexanoic acid production is seen in regioselectivity assays, where (R)-2-ethylhexanoic acid comprises 50% of total products while (S)-2-ethylhexanoic acid comprises only 13%. (R)- and (S)-2-ethylhexanol exhibit similar characteristics with respect to the amount of oxygen and reducing equivalents consumed. however, with (S)-2-ethylhexanol turnover producing more water than the (R)-enantiomer. Crystallographic studies of P450cam with (R)- or (S)-2-ethylhexanoic acid suggest that the (R)-enantiomer binds in a more ordered state. These results indicate that wild-type P450cam displays stereoselectivity toward 2-ethylhexanoic acid synthesis, providing a platform for rational active site design.
