Mutagenesis of the MAT alpha Secretion Signal in Pichia Pastoris
Poster Number
42
Format
Poster Presentation
Faculty Mentor Name
Geoffrey Lin-Cereghino
Faculty Mentor Department
Biological Sciences
Additional Faculty Mentor Name
Joan Lin-Cereghino
Abstract/Artist Statement
The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins is initially fused to the Saccharomyces cerevisiae alpha mating factor secretion signal. The alpha mating factor secretion signal acts like an address label that should direct the protein to be secreted out of the cell. We surmised the removal of certain regions of amino acids should facilitate HRP protein secretion by increasing the flexibility of the MAT alpha loop region. Using bioinformatics, the Lin-Cereghino lab has created a model of the secretion signal. Using this model as a guide, we have created a series of mutated secretion signal sequences in order to comprehend how this cellular address label functions and to see if we can achieve higher levels of secretion. The results allowed us to understand the contribution of each segment in directing protein secretion. Our findings raise the possibility that the secretory function of this alpha mating factor signal can be further enhanced in the future.
Location
DeRosa University Center, Ballroom
Start Date
20-4-2013 1:00 PM
End Date
20-4-2013 3:00 PM
Mutagenesis of the MAT alpha Secretion Signal in Pichia Pastoris
DeRosa University Center, Ballroom
The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for industrial and research purposes. In order to secrete proteins for easier purification from the extracellular medium, the coding sequence of recombinant proteins is initially fused to the Saccharomyces cerevisiae alpha mating factor secretion signal. The alpha mating factor secretion signal acts like an address label that should direct the protein to be secreted out of the cell. We surmised the removal of certain regions of amino acids should facilitate HRP protein secretion by increasing the flexibility of the MAT alpha loop region. Using bioinformatics, the Lin-Cereghino lab has created a model of the secretion signal. Using this model as a guide, we have created a series of mutated secretion signal sequences in order to comprehend how this cellular address label functions and to see if we can achieve higher levels of secretion. The results allowed us to understand the contribution of each segment in directing protein secretion. Our findings raise the possibility that the secretory function of this alpha mating factor signal can be further enhanced in the future.