Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocosReportar como inadecuado




Cloning and Characterization of Farnesyl Diphosphate Synthase Gene Involved in Triterpenoids Biosynthesis from Poria cocos - Descarga este documento en PDF. Documentación en PDF para descargar gratis. Disponible también para leer online.

Guangdong VTR Bio-Tech Co., Ltd., Zhuhai 519060, Guangdong, China



These authors contributed equally to this work.





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Abstract Poria cocos P. cocos has long been used as traditional Chinese medicine and triterpenoids are the most important pharmacologically active constituents of this fungus. Farnesyl pyrophosphate synthase FPS is a key enzyme of triterpenoids biosynthesis. The gene encoding FPS was cloned from P. cocos by degenerate PCR, inverse PCR and cassette PCR. The open reading frame of the gene is 1086 bp in length, corresponding to a predicted polypeptide of 361 amino acid residues with a molecular weight of 41.2 kDa. Comparison of the P. cocos FPS deduced amino acid sequence with other species showed the highest identity with Ganoderma lucidum 74%. The predicted P. cocos FPS shares at least four conserved regions involved in the enzymatic activity with the FPSs of varied species. The recombinant protein was expressed in Pichia pastoris and purified. Gas chromatography analysis showed that the recombinant FPS could catalyze the formation of farnesyl diphosphate FPP from geranyl diphosphate GPP and isopentenyl diphosphate IPP. Furthermore, the expression profile of the FPS gene and content of total triterpenoids under different stages of development and methyl jasmonate treatments were determined. The results indicated that there is a positive correlation between the activity of FPS and the amount of total triterpenoids produced in P. cocos. View Full-Text

Keywords: farnesyl diphosphate synthase; methyl jasmonate; Poria cocos; triterpenoids farnesyl diphosphate synthase; methyl jasmonate; Poria cocos; triterpenoids





Autor: Jianrong Wang †,* , Yangyuan Li † and Danni Liu

Fuente: http://mdpi.com/



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