Cloning of gene encoding phytase included hexahistidine-tag from the Peniophora lycii in yeast

Aim and Phytic acid is an important mineral compound which can be preserved in plants. Phytase (E.C. 3.1.3.26 or E.C. 3.1.3.8) has been used as a cereal feed additive that can enhance the phosphorus and mineral absorption in monogastric animals food; so it can reduce the phosphorus output into the environment. In this research, Cloning of gene encoding phytase containing hexahistidine-tag was conducted in Escherichia coli and then yeast for purification of the produced enzyme after expression in this yeast. Sequence of protein encoding Peniophora lycii fungal phytase, was obtained from protein data bank, designed and then synthesized according to the expression codon usage of the yeast. A piece of DNA containing hexahistidine-tag was added into the pFPMTMF 14α'> shuttle vector fallowed by transformation through electroporation method into the expression yeast. Recombinant DNA encoding phytase containing hexahistidine-tag was produced. Presence of gene in bacteria and yeast was proved with double digest and PCR methods. Successful cloning of fungal phytase gene containing hexahistidine-tag was conducted in Escherichia coli bacteria and then yeast for high level expression and purification of this enzyme.