Paper Infomation
Cloning And Characterization Of A Geranylgeranyl Diphosphate Synthase Gene From Taxus Chinensis
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Author: Junhao Shen, Yangyang Qiao, Yongqin Chen, Jungui Dai, Qiqi Chen, Zhifan Yang, Xuzhi Ruan, Ronghua Wei
Abstract: Taxol (Paclitaxel) is a diterpene from Taxus species and has been used in treatment of various kinds of cancers. Geranylgeranyl diphosphate synthase (GGPPS) catalyzes the formation of geranylgeranyl diphosphate (GGPP, the common precursor for diterpenes and plays a key role in taxol biosynthesis. Here we report a functional GGPPS gene from Taxus chinensis (designated TcGGPPS). TcGGPPS is an intron free gene and has a 1,182-bp open reading frame encoding a polypeptide of 393 amino acid residues with a calculated molecular mass of 42.63 kDa and an isoelectric point of 5.58. The catalytic activity of TcGGPPS for production of GGPP was verified by a color enhancement assay in the Escherichia coli cells harboring plasmid pAC-BETA. Multiple sequence alignment indicates that TcGGPPS is a little different in sequence from the functional GGPPS genes from other Taxus species such as T. canadensis, T. media and T. wallichiana, which are almost identical to each other. Protein structure prediction by using bioinformatics reveals that TcGGPPS consists of 52.2% α-helix, 10.9% extended strand, 8.4% β-turn and 28.5% random coil, and has a three-dimensional structure highly similar to the structurally known Sinapis alba GGPPS. In silicon predictions also demonstrate that TcGGPPS has a plastid-targeting peptide at the N-terminus, suggesting it is responsible for the synthesis of GGPP in plastids.
Keywords: Taxus chinensis; Geranylgeranyl Diphosphate Synthase; Gene Cloning; Taxol Biosynthesis
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