Production of extracellular recombinant phytase in yeast and its application in animal feed as enzyme supplement.

The expression of recombinant proteins in Saccharomyces cerevisiae has been practiced for last two decades. Although different expression systems have been made for specific protein production, the system with broad applicability was not developed so far. The research proposed in this project will focus on production of extracellular phytase enzyme in yeast S. cerevisiae and its potential application in animal feed. 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 extracellular expression 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 producing phytase enzyme in the media. 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 produced recombinant 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 Khulna University Research Cell, Khulna University, Bangladesh for two years starting from 2021 to 2023.

• Research Project