Following evolution's lead to a single residue switch for diterpene synthase product outcome

被引:143
作者
Xu, Meimei [1 ]
Wilderman, P. Ross [1 ]
Peters, Reuben J. [1 ]
机构
[1] Iowa State Univ, Dept Biochem Biophys & Mol Biol, Ames, IA 50011 USA
关键词
enzyme specificity; natural product biosynthesis; terpene synthase; biochemical evolution;
D O I
10.1073/pnas.0611454104
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
There have been few insights into the biochemical origins of natural product biosynthesis from primary metabolism. Of particular interest are terpene synthases, which often mediate the committed step in particular biosynthetic pathways so that alteration of their product outcome is a key step in the derivation of novel natural products. These enzymes also catalyze complex reactions of significant mechanistic interest. Following an evolutionary lead from two recently diverged, functionally distinct diterpene synthase orthologs from different subspecies of rice, we have identified a single residue that can switch product outcome. Specifically, the mutation of a conserved isoleucine to threonine that acts to convert not only the originally targeted isokaurene synthase into a specific pimaradiene synthase but also has a much broader effect, which includes conversion of the ent-kaurene synthases found in all higher plants for gibberellin phytolhormone biosynthesis to the production of pimaradiene. This surprisingly facile switch for diterpene synthase catalytic specificity indicates the ease with which primary (gibberellin) metabolism can be subverted to secondary biosynthesis and may underlie the widespread occurrence of pimaradiene-derived natural products. In addition, because this isoleucine is required for the mechanistically more complex cyclization to tetracyclic kaurene, whereas substitution with threonine "short-circuits" this mechanism to produce the "simpler" tricyclic pimaradiene, our results have some implications regarding the means by which terpene synthases specify product outcome.
引用
收藏
页码:7397 / 7401
页数:5
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