Bioinformatic analysis and Protein Properties of Delta ۶ desaturase Enzyme Cloned in Pichia pastoris

Background: Fungi are the best microorganism to production of essential fatty acids, but due tothe high-volume industrial production, gene cloning in the yeast was used commonly.Materials and Methods: In this study, cloning of delta ۶ desaturase gene obtained from fungiMucor rouxii DSM۱۱۹۴ in the yeast Pichia pastorisGS۱۱۵ was performed and the expression ofGLA production as omega ۶ fatty acid in the recombinant strain was confirmed. The completesequence of the cloned delta ۶ desaturase protein was examined using Expasy browser and thephylogenetic tree of the delta ۶ desaturase gene was plotted by CLC software. The finalrecombinant vector map was drawn by Snap gene viewer software and the Delta ۶ desaturasegene, which is located between the restriction enzymes EcoRI and XhoI was confirmed.Results: By using the CFSSP server, the second structure of cloned protein was predicted.Investigations performed on the PDB server revealed that the crystallographic structure of thedelta ۶ desaturase enzyme had not yet been studied or recorded. Study showed the cloned deltadesaturase enzyme containing ۵۵۶ amino acids. The protein alignment of this study with thereference protein in the NCBI gene bank shows that these two proteins are different in threepositions ۸۴, ۱۱۱ and ۲۷۱.Conclusion: According to the studies, it was found that the parts that have been altered in thisprotein are all related to extracellular sequences, and the sequences that pass through themembrane and inside the cell are quite similar to the reference protein.

Keywords:

Cloning , Delta۶ desaturase , Pichia pastoris , Bioinformatic analysis