Production of pure and high8208;yield client proteins is an important technology that attends the needs for industrial applications of enzymes as well as basic experiments such as protein crystallization. Client protein expression platforms are available in Escherichia coli and the methylotrophic Pichia pastoris that result in proteins released to the intracellular cell extract and extracellular medium, respectively. Fungi are utilized in industrial protein production because of their ability to secrete large quantities. In this study we engineer a high8208;expression8208; secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose overexpression, production time and pH8208;dependent protein stability in the medium. We describe five client proteins that accumulated 508208;100 mg of protein per liter and only one protein was secreted at low quantities. We also test a recyclable genetic marker that allowed secretion of multiple client proteins, enabling the design of an enzyme activity set.
Full conference title:
11 th European Conference on Fungal Genetics
- ECFG 11th (2012)