Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on March 31, 2006; 10.1104/pp.106.077008

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Received January 12, 2006
Returned for revision February 7, 2006
Accepted March 22, 2006

Maize cDNAs expressed in endosperm encode functional farnesyl diphosphate synthase with geranylgeranyl diphosphate synthase activity

Miguel Cervantes-Cervantes , Cynthia E. Gallagher , Changfu Zhu , and Eleanore T. Wurtzel *

Department of Biological Sciences, Lehman College, The City University of New York (CUNY), 250 Bedford Park Boulevard West, Bronx, New York 10468 and The Graduate School and University Center-CUNY, 365 Fifth Avenue, New York, New York 10016

* Corresponding author; email: wurtzel{at}lehman.cuny.edu.

Isoprenoids are the most diverse and abundant group of natural products. In plants, farnesyl diphosphate (FPP) and geranylgeranyl diphosphate (GGPP) are precursors to many isoprenoids having essential functions: Terpenoids and sterols are derived from FPP, whereas gibberellins, carotenoids, casbenes, taxenes and others originate from GGPP. The corresponding synthases (FPPS and GGPPS) catalyze, respectively, the addition of two and three isopentenyl diphosphate molecules to dimethylallyl diphosphate. Maize (Zea mays L. cv. B73) endosperm cDNAs encoding isoprenoid synthases were isolated by functional complementation of Escherichia coli cells carrying a bacterial gene cluster encoding all pathway enzymes needed for carotenoid biosynthesis, except for GGPPS. This approach indicated that the maize gene products were functional GGPPS enzymes. Yet, the predicted enzyme sequences revealed FPPS motifs and homology with FPPS enzymes. In vitro assays demonstrated that indeed these maize enzymes produced both FPP and GGPP and that the N-terminal sequence affected the ratio of FPP/GGPP. Their functionality in E. coli demonstrated that these maize enzymes can be coupled with a metabolon to provide isoprenoid substrates for pathway use, and suggests that enzyme bifunctionality can be harnessed. The maize cDNAs are encoded by a small gene family whose transcripts are prevalent in endosperm beginning mid-development. These maize cDNAs will be valuable tools for assessing the critical structural properties determining prenyl transferase specificity and in metabolic engineering of isoprenoid pathways, especially in cereal crops.




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