High-Yield Secretion of Multiple Client Proteins in Aspergillus

Fernando Segato, André R.L. Damásio, Thiago A. Gonçalves, Rosymar C. de Lucas, Fabio M. Squina, Stephen R. Decker, Rolf A. Prade

Research output: Contribution to journalArticlepeer-review

71 Scopus Citations

Abstract

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/L 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.

Original languageAmerican English
Pages (from-to)100-106
Number of pages7
JournalEnzyme and Microbial Technology
Volume51
Issue number2
DOIs
StatePublished - 15 Jul 2012

NREL Publication Number

  • NREL/JA-2700-56073

Keywords

  • 5-FOA
  • CAZy
  • CMC
  • DNS
  • Fungi
  • HEPES
  • Heterologous expression and secretion of multiple proteins
  • MM
  • PCR
  • Protein production
  • Protein secretion
  • SDS-PAGE

Fingerprint

Dive into the research topics of 'High-Yield Secretion of Multiple Client Proteins in Aspergillus'. Together they form a unique fingerprint.

Cite this