Biotechnology and genetics
Mohammad Javad Habibzadeh; Seyed Mahdi Ziaratnia; Ebrahim Dorani-Uliaie
Abstract
Saffron is the most expensive spice in the world. The economic value of saffron is due to the existence of apocarotenoids in its stigma. Therefore, the isolation and characterization of genes involving in apocarotenoids metabolism is on particular importance. In this research, beacause the importance ...
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Saffron is the most expensive spice in the world. The economic value of saffron is due to the existence of apocarotenoids in its stigma. Therefore, the isolation and characterization of genes involving in apocarotenoids metabolism is on particular importance. In this research, beacause the importance of CsUGT gene in crocin biosynthesis, it was isolated, cloned the E.coli strain DH5α. The full length gene was sequenced and registered in the NCBI. In order to characterize CsUGT gene, first the protein sequence was obtained, and then the physical and chemical characteristics and physiology of the CsUGT protein were analyzed by Protparam, SOPMA, ProtScale, Pfam, ProtComp, SignalP, TMHMM and ChloroP servers and tools. Also, using the Swiss-Model server, the 3D structure of this protein was investigated and Ramachandran diagram was drawn to validate the 3D drawn model structure. According to the results, CsUGT protein with 462 amino acids has the conserved sequence of glycosyltransferase family proteins and was identified as a polar protein, stable at high temperatures and without hydrophobic domain. CsUGT protein has no peptide signal or binding signals and has a cytoplasmic location. This research made it possible to isolate the CsGTS gene of saffron and optimized the conditions of its transfer into a vector. In addition, the results of CsUGT protein structure analysis provide the basis for future functional studies and can also provide valuable information regarding the behavior and reaction of this enzyme in the synthesis of saffron apocarotenoids. In addition, these results can be useful in the future programs of Iranian saffron genetic modification.
Mohammad Javad Habibzadeh; Ebrahim Dorani; Seyed Mahdi Ziaratnia; Mostafa Valizadeh
Abstract
Saffron is one of the most expensive spices and natural colors used in various food, pharmaceutical and cosmetic industries. In recent years, a family of enzymes that digest carotenoid substrates into double bonds are identified and introduced in plants. This family is of enzymes Carotenoid Cleavage ...
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Saffron is one of the most expensive spices and natural colors used in various food, pharmaceutical and cosmetic industries. In recent years, a family of enzymes that digest carotenoid substrates into double bonds are identified and introduced in plants. This family is of enzymes Carotenoid Cleavage Dioxygenase (CCD) enzymes. In this study, two isoforms of this gene were cloned and sequenced due to the importance of CCD genes in biosynthesis of apocarotenoids. Bioinformatics analyses including phylogenetic relationships and protein structures were evaluated. 3D modeling of these proteins was done by homologous modeling and using the Swiss Model database after selecting the appropriate pattern. The Ramachandran plot was drawn in order to validate the structure of the 3D model. The results show that the two CCD4a and CCD4b isoforms have both exons and one intron. In silico analysis, the physicochemical properties of CsCCD4a and CsCCD4b proteins also show that the proteins derived from these two isoforms are similar in terms of molecular weight, amino acids, isoelectric points, aliphatic index, instability index and solubility. The results of study of 3D structures resulted in proposal of similar structures for two isoforms. The results of this study can provide valuable information on the behavior and response of CCD4 enzyme in the pathway for synthesis of apricotines in saffron, and these results can be useful in future protein engineering programs.
