Supersecretion in Pichia pastoris mutant strains

• Main Authors: Agrawal, Ruchika, Afnan, Mina, Ha, Stephanie, Park, Sangwook, Scheel, Jacob
• Format: Text
• Language: unknown
• Published: Scholarly Commons 2017
• Subjects: Biology; Antarc*; Antarctica
• Online Access: https://scholarlycommons.pacific.edu/purcc/2017/events/82

Pichia pastoris is a methylotropic yeast that is commonly used for the secretion of large amounts of heterologous protein products. Continuous progress towards increasing secretion of protein products in Pichia pastoris is important for commercial and academic value. While supersecretor mutant strains of Pichia pastoris have been discovered, the mechanism behind supersecretion is unclear. Supersecretion can be attributed to defects in cell wall integrity or to a difference in protein secretion pathway in the mutants. The goal of this project is to use the reporter enzyme Candida antarctica lipase B (CalB) and secretory leukocyte protease inhibitor (SLPI) to test if supersecretion is due to defects in cell wall integrity. For this purpose we have constructed the plasmids pGAPHISαB, pGAPHISαB–CalB and pGAPHISαB-SLPI. The results will allow increased understanding of the mode of supersecretion of the mutant strains.