Fungal protease: Production, purification and compatibility with laundry detergents and their wash performance
Journal
Journal of the Taiwan Institute of Chemical Engineers
Journal Volume
42
Journal Issue
2
Pages
298-304
Date Issued
2011
Author(s)
Abstract
An extracellular serine protease producing fungi were isolated from the effluent sample collected from a sago industry in Salem and was identified as Graphium putredinis and Trichoderma harzianum. Intergenerically developed fusant produced high amounts of protease in soya bean meal amended minimal medium than the parents, G. putredinis and T. harzianum. The enzyme was purified by Sephadex G 100 column chromatography. The proteases of G. putredinis and T. harzianum had optimum pH and temperature of 7.0-8.0 and 50-60°C respectively. At 37 and 60°C, the parental proteases were respectively stable for 1 day and 15min and the fusant was stable for 2 days and 10min. The Km and Vmax were 0.65, 1.25 and 0.40mg/ml and 2.00, 1.60 and 2.60IU/mg protein for G. putredinis, T. harzianum and fusant respectively with a molecular weight of 31, 20 and 33kDa. The effect of metal ions showed that, at 5mM concentration of Hg2+, the residual protease activity of parent and fusant was 5.47, 2.48 and 9.76% respectively; Cu2+, Ca2+ and Zn2+ did not greatly affect the enzyme activity. In 5mM EDTA showed residual activity between 60.76 and 85.66%. PMSF (2mM) completely inhibited the protease activity of all the three fungi studied. All the three fungal enzymes retained maximum residual activity of 66.00, 63.80 and 76.74% with the commercial detergent Rin Advanced. With Kite, all the enzymes had more or less same level of residual activity (54-55%). With SDS and sodium perborate (0.2%) the residual activities were 58.25-73.82 and 61.58-70.24% respectively. Wash performance analysis revealed that fusant protease with Rin Advanced at 60°C yielded good result. © 2010 Taiwan Institute of Chemical Engineers.
Type
journal article