Title

Mat Alpha Leader Analysis for Pichia Pastoris Secretion

Poster Number

30

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Geoff 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 modern medical and research goals. 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. This “MAT alpha” signal acts like an address label that should direct the protein 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 as a guide for a series of mutated sequences in order to comprehend how this region functions and to further examine key mutations. Mutation delta 57-70 has shown very high secretion (150% of wild type) and mutation delta 61-70 has shown low secretion (80% of wild type). Our lab has been working to determine the secretion of mutations delta 58-70, 59-70, and 60- 70 to narrow down the important amino acids in mass secretion for MAT alpha. The findings from this experiment suggest which amino acids play a key role in higher efficiency in the MAT alpha. Our interpretations raise the possibility of creating more efficient processes of secreting proteins that are vital to products that fight diseases such as cancer, hepatitis B&C, and diabetes.

Location

DeRosa University Center, Ballroom

Start Date

30-4-2016 10:00 AM

End Date

30-4-2016 12:00 PM

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Apr 30th, 10:00 AM Apr 30th, 12:00 PM

Mat Alpha Leader Analysis for Pichia Pastoris Secretion

DeRosa University Center, Ballroom

The methylotrophic yeast, Pichia pastoris, has been genetically engineered to produce many heterologous proteins for modern medical and research goals. 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. This “MAT alpha” signal acts like an address label that should direct the protein 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 as a guide for a series of mutated sequences in order to comprehend how this region functions and to further examine key mutations. Mutation delta 57-70 has shown very high secretion (150% of wild type) and mutation delta 61-70 has shown low secretion (80% of wild type). Our lab has been working to determine the secretion of mutations delta 58-70, 59-70, and 60- 70 to narrow down the important amino acids in mass secretion for MAT alpha. The findings from this experiment suggest which amino acids play a key role in higher efficiency in the MAT alpha. Our interpretations raise the possibility of creating more efficient processes of secreting proteins that are vital to products that fight diseases such as cancer, hepatitis B&C, and diabetes.