Protein Secretion Machines

High-yield Secretion of Multiple Client Proteins in Aspergillus

Production of pure and high-yield client proteins is an important technology that addresses the need for industrial applications of enzymes as well as scientific experiments in protein chemistry and crystallization.  Fungi are utilized in industrial protein production because of their ability to secrete large quantities of proteins.  In this study, we engineered a high-expression-secretion vector, pEXPYR that directs proteins towards the extracellular medium in two Aspergillii host strains, examine the effect of maltose-induced over-expression and protein secretion as well as time and pH-dependent protein stability in the medium.  We describe five client proteins representing a core set of hemicellulose degrading enzymes that accumulated up to 50-100 mg/liter of protein.  Using a recyclable genetic marker that allows serial insertion of multiple genes, simultaneous hyper-secretion of three client proteins in a single host strain was accomplished.



Cell & Molecular Biology
ONLINE  Cell & Molec. Biol.
Medical Mycology
Eukaryotic Genetics
Biomass Technology
Biomass Degradation
High Temp. Enzymes
Protein expression systems

Molecular Genetics
Gene Silencing
Stress Responses
Gene Regulation

DNA Bioinformatics
Structural Protein Modeling
Physical Mapping