Surface display of phytase in the yeast cell wall and its application in animal feed as enzyme supplement.

The expression of recombinant proteins immobilised at the cell surface of Saccharomyces cerevisiae, termed yeast surface display (YSD), has now been practiced for last two decades. Although different surface display systems have been made for specific purposes, the system with broad applicability was not developed so far. The research proposed in this project will focus on production of phytase enzyme in yeast S. cerevisiae and its potential application in biotechnology. Using knowledge of molecular mechanisms and techniques of recombinant plasmid construction, a novel plasmid for recombinant protein production will be constructed that would enable easy insertion of the gene coded for phytase enzyme in the genetic cassette consisting of a host promoter, signal sequence for directing the protein into the secretory pathway and genetic tags to enable easy detection of the recombinant protein. The genetic cassette will allow easy insertion of any gene of interest by restriction cloning method and concomitant surface display of recombinant protein in yeast. The Aspergillus niger gene PhyA coding for phytase will be inserted in the prepared genetic cassette in order to construct yeast strain diplaying phytase enzyme at the cell surface. The functionality of genetic construct will be checked by Western blot analysis of protein synthesized after induction on galactose. The activity of the recombinant enzyme will be analysed by UV spectrophotometric analysis. The surface displayed phytase enzyme will be further characterized and compared with the commercially available phytase.

I am the principle investigator of this project. The research project is funded by The Ministry of Education, Bangladesh for three years starting from 2021 to 2024.

• Research Project