Optimisation of cellobiose dehydrogenase production by the fungus Sclerotium (Athelia) rolfsii.
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.
Ludwig
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Division of Biochemical Engineering, University of Agricultural Sciences Vienna, Muthgasse 18, 1190 Vienna, Austria.
Haltrich
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D000596:Amino Acids / Q000145:classification / Q000378:metabolism; D002237:Carbohydrate Dehydrogenases / Q000096:biosynthesis*; D002244:Carbon / Q000378:metabolism; D003470:Culture Media; D007218:Industrial Microbiology; D009584:Nitrogen / Q000378:metabolism; D020072:Polyporales / Q000201:enzymology* / Q000254:growth & development; D011480:Protease Inhibitors / Q000378:metabolism; D013997:Time Factors
D000596:Amino Acids; D003470:Culture Media; D011480:Protease Inhibitors; D002244:Carbon; D002237:Carbohydrate Dehydrogenases; C019859:cellobiose-quinone oxidoreductase; D009584:Nitrogen