Heterologous production of Aspergillus fumigatus keratinase in Pichia pastoris

摘要: Aspergillus fumigatus Fresenius was previously shown to grow in mineral medium containing chicken feather flour as carbon and nitrogen source. Substantial proteolytic keratin-degrading activity present the culture supernatant after 24–72 h of growth at 42 °C. The keratinase successfully purified by a single ion exchange chromatographic procedure had molecular mass 31 kDa determined SDS–PAGE. cDNA expressed Pichia pastoris cells recombinant clones were be able produce substantial caseinolytic, azo-keratinolytic keratinolytic activities. SDS–PAGE Western-blotting analysis using antibody against A. showed presence protein supernatants several P. cells. This corresponding that keratinase. enzyme production profile theP. produced an increasing amount activities over 72 period. Dry weight determination indicated 10% keratin hydrolysed 24 incubation period with 510 U (caseinolytic activity)