Optimisation of cellobiose dehydrogenase production by the fungus Sclerotium (Athelia) rolfsii
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The phytopathogenic fungus Sclerotium (Athelia) rolfsii CBS 191.62 is a very efficient producer of the hemoflavoprotein, cellobiose dehydrogenase (CDH), forming up to 225 mg l−1 (15,000 units cytochrome c activity l−1) of this protein, which is of biotechnological interest for sensors, biocatalysis and bioremediation. Both cellulose as inducing substrate and the use of a rich medium containing increased concentrations of peptone from meat or suitable amino acids are important for attaining high CDH yields. CDH, containing a protease-sensitive linker region, can be cleaved by endogenous proteases into a catalytically active flavin fragment and an inactive heme domain. By using increased concentrations of peptone, or certain amino acids such as valine or leucine, or by adding exogenous protease inhibitors, this cleavage can be almost completely inhibited, so that more than 95% intact CDH is obtained under optimised culture conditions. When using non-inhibitory amino acids, e.g. glutamine or lysine, in the medium, more than 80% of the total cellobiose-oxidising activity can be attributed to the flavin fragment.
KeywordsPeptone Protease Activity Cellobiose DCIP Extracellular Protease Activity
We thank Andrea Salamon and Cindy Lorenz for excellent technical assistance. This work was supported by the Austrian Research Foundation, project FWF P14537. The experiments performed comply with the current laws of Austria.
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