Title

The Search for Super-Secreters and the Secret to their Superpowers

Poster Number

12

Format

Poster Presentation

Abstract/Artist Statement

Pichia pastoris is a strain of yeast often used by academic and commercial laboratories as a source for heterologous protein expression. However, despite its ability to produce specific proteins, P. pastoris is unable to efficiently express and secrete certain proteins, such as the - galactosidase enzyme, in substantial amounts. Prior to this current project, a random mutagenesis was conducted using Restriction Enzyme Mediated Integration, 18 strains were isolated and found to be potential super-secreters of -galactosidase. Our objective was to unravel the secret identity of the disrupted gene in order to illuminate the secretory mechanisms. Genomic DNA with the mutation from super-secreter strains AH14-4 and AH8-2 was isolated, sequenced, and analyzed. Through BLAST analysis, the disrupted gene in the mutant strain AH8-2 may be responsible for a golgi matrix protein, while the disrupted gene in mutant strain AH14-4 may be a non-essential subunit of the exocyst complex. This project provides knowledge about P. pastoris’s secretory machinery which could lead to improvements which make P. pastoris a more "powerful" system for heterologous expression.

Location

DeRosa University Center, Ballroom B

Start Date

2-5-2009 1:00 PM

End Date

2-5-2009 3:00 PM

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May 2nd, 1:00 PM May 2nd, 3:00 PM

The Search for Super-Secreters and the Secret to their Superpowers

DeRosa University Center, Ballroom B

Pichia pastoris is a strain of yeast often used by academic and commercial laboratories as a source for heterologous protein expression. However, despite its ability to produce specific proteins, P. pastoris is unable to efficiently express and secrete certain proteins, such as the - galactosidase enzyme, in substantial amounts. Prior to this current project, a random mutagenesis was conducted using Restriction Enzyme Mediated Integration, 18 strains were isolated and found to be potential super-secreters of -galactosidase. Our objective was to unravel the secret identity of the disrupted gene in order to illuminate the secretory mechanisms. Genomic DNA with the mutation from super-secreter strains AH14-4 and AH8-2 was isolated, sequenced, and analyzed. Through BLAST analysis, the disrupted gene in the mutant strain AH8-2 may be responsible for a golgi matrix protein, while the disrupted gene in mutant strain AH14-4 may be a non-essential subunit of the exocyst complex. This project provides knowledge about P. pastoris’s secretory machinery which could lead to improvements which make P. pastoris a more "powerful" system for heterologous expression.