Author/Authors :
-، - نويسنده Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan, IR Iran Abbasi Kajani, Abolghasem , -، - نويسنده Department of Biochemistry, School of Pharmacy and Pharmaceutical Sciences, Bioinformatics Research Center, Isfahan University of Medical Sciences, Isfahan, IR Iran Mofid, Mohamma Reza , -، - نويسنده Ramin Agricultural and Natural Resources University, Mollasani, Ahwaz, IR Iran Alami Saeid, Khalil
Abstract :
Background: Taxol is one of the most important anti-cancer drugs, which is obtained from yew trees (Taxus sp.). T+he fist step in side chainassembly of taxol is catalyzed by phenylalanine aminomutase, which converts α-phenylalanine to β-phenylalanine.Objectives: In this study, for the fist time, we report on the cloning, preliminary expression and characterization of a full-length gene andcDNA encoding phenylalanine aminomutase from Taxus baccata L.Materials and Methods: Comparison of the full-length gene with other ones identifid from the Taxus species showed high similarity,particularly with Taxus x media.Results: The results showed that the expression level of this gene in Taxus baccata is very low and therefore this enzymatic step could be arate limiting step in the taxol biosynthesis pathway. Successful amplifiation of the cDNA was only obtained from RNA samples isolated frommethyl jasmonate elicited suspension cells of Taxus baccata. The cloned cDNA contained a 2064 bp open reading frame encoding a proteincomposed of 687 amino acids. Sequence comparison analysis revealed that the gene is very similar (98 - 99 %) with respect to the nucleotideand amino acid sequences in diffrent Taxus species and also share the signature active site motif (175ASG177).Conclusions: The predicted structure of TbPAM was analyzed using bioinformatic tools. The results indicated that the protein has similaroverall folding to tyrosine aminomutase.
