Characterising extra-cellular keratinase secreted by the ascomycetous fungus, A16 Aspergillus giganteus. Emphasis on properties, conditions, applicationsдипломная работа (Бакалавр)
Организация, в которой проходила защита:
Swansea University
Год защиты:2022
Аннотация:The fungal species, Aspergillus giganteus was chosen to be investigated, to see, if it can be a potential producer of extra-cellular keratinases capable of degrading keratinous waste. Many Aspergillus species have been found to secrete keratinases with beneficial characteristics applicable in many industries. In this investigation, an extra cellular keratinise was successfully purified from the protein crude extract of Aspergillus giganteus obtained under solid-state fermentation using chicken feathers as a substrate. The keratinase was then separated and purified to homogeneity by performing isoelectric focusing and SDS-Polyacrylamide gel electrophoresis. The molecular weight of the keratinase was recognised as 27 kDa. 10 synthetic substrates were used to determine substrate specificity, of which the Subtilisin A-specific substrate, Z-Ala-Ala-Leu-pNA, showed greatest activity. Its full inhibition to serine protease inhibitor, PMSF and partial inhibition by chelating agent EDTA, categorised the enzyme as being a serine protease which partially uses metal ions during substrate hydrolysis. The optimum conditions for A16 keratinase activity and stability was found at 30C with a pH of 8.0. Its biochemical properties give the keratinase potential to be used in hygiene (in detergents and cleaning agents), biomedical (as a way of treating blood-clot disorders) and agricultural industries (increasing meat quality and turning waste products into highly nutritious materials)