通过香叶基香叶基还原酶的结构导向修饰构建定制的类异戊二烯产物。
Constructing tailored isoprenoid products by structure-guided modification of geranylgeranyl reductase.
作者信息
Kung Yan, McAndrew Ryan P, Xie Xinkai, Liu Charlie C, Pereira Jose H, Adams Paul D, Keasling Jay D
机构信息
Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA; Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Department of Chemistry, Bryn Mawr College, 101 North Merion Avenue, Bryn Mawr, PA 19010, USA.
Physical Biosciences Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Road, Berkeley, CA 94720, USA; Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA.
出版信息
Structure. 2014 Jul 8;22(7):1028-36. doi: 10.1016/j.str.2014.05.007. Epub 2014 Jun 19.
The archaeal enzyme geranylgeranyl reductase (GGR) catalyzes hydrogenation of carbon-carbon double bonds to produce the saturated alkyl chains of the organism's unusual isoprenoid-derived cell membrane. Enzymatic reduction of isoprenoid double bonds is of considerable interest both to natural products researchers and to synthetic biologists interested in the microbial production of isoprenoid drug or biofuel molecules. Here we present crystal structures of GGR from Sulfolobus acidocaldarius, including the structure of GGR bound to geranylgeranyl pyrophosphate (GGPP). The structures are presented alongside activity data that depict the sequential reduction of GGPP to H6GGPP via the intermediates H2GGPP and H4GGPP. We then modified the enzyme to generate sequence variants that display increased rates of H6GGPP production or are able to halt the extent of reduction at H2GGPP and H4GGPP. Crystal structures of these variants not only reveal the structural bases for their altered activities; they also shed light onto the catalytic mechanism employed.
古菌香叶基香叶基还原酶（GGR）催化碳 - 碳双键的氢化反应，以生成该生物体不寻常的类异戊二烯衍生细胞膜的饱和烷基链。类异戊二烯双键的酶促还原反应，对于天然产物研究人员以及对微生物生产类异戊二烯药物或生物燃料分子感兴趣的合成生物学家来说，都具有相当大的吸引力。在此，我们展示了嗜酸热硫化叶菌（Sulfolobus acidocaldarius）GGR的晶体结构，包括与香叶基香叶基焦磷酸（GGPP）结合的GGR结构。这些结构与活性数据一同呈现，这些数据描述了GGPP通过中间体H2GGPP和H4GGPP逐步还原为H6GGPP的过程。然后，我们对该酶进行修饰，以生成显示出更高H6GGPP生成速率或能够在H2GGPP和H4GGPP处停止还原程度的序列变体。这些变体的晶体结构不仅揭示了其活性改变的结构基础；还阐明了所采用的催化机制。
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