A Collaborative Proposal: Engineering Disulfide Formation Kinetics to Enhance Heterologous Secretion in Saccharomyces Cerevisiae

Massachusetts Institute Of Technology, Cambridge MA

Investigators

Abstract

The secretion of heterologous proteins that contain disulfide bonds is not at its full potential. Evidence suggests that the regulation of the redox state of the endoplasmic reticulum may be a bottleneck. A multifaceted approach is proposed to probe and manage the redox state in yeast in order to investigate the bottleneck hypothesis further and to ultimately increase secretion efficiency. The four elements of the proposed work are: (1) amplify the products of the individual genes now thought to be involved redox management to assess the effect, (2) use display methods to uncover possibly missing pathway elements, (3) add redox altering compounds to the growth medium to alter redox state, and (4) develop a tachyplesin I substrate-based diagnostic system.