Filamentous fungi have a very efficient protein-production capacity which make them suitable host organisms for the overproduction of commercially interesting homologous and heterologous proteins. The overall efficiency of an enzyme production process is influenced by the production yield (fermentation) and purification yield (Down Stream Processing). Unfortunately, since every protein is different, in many cases production and purification protocols and strategies must be developed for each individual protein. In E. coli the fusion of proteins to oligohistidine tags followed by affinity chromatography is a very common protein purification strategy. As far as we know, the use of His-tags in the extracellular production and purification of heterologous proteins in Aspergillus has not yet been demonstrated. Recently, we successfully produced Arthromyces ramososus peroxidase (ARP) in Aspergillus niger under control of the inuE (exo-inulinase) expression signals. To allow fast and easy purification we introduced N- and C-terminal His6-tags, respectively. Extracellular peroxidase activity could be measured and was obtained with both, N- and C-terminal His6-tagged ARP. The ability to purify the different His6-tagged proteins by affinity chromatography is under investigation.
Full conference title:
10th EUROPEAN CONFERENCE ON FUNGAL GENETICS
- ECFG 10th (2010)