ISOLATION, EXTRACTION, PURIFICATION AND CHARACTERIZATION OF ALKALINE PROTEASE FROM NEUROSPORA CRASSA AND ENZYME ASSAY
About Authors:
Anuradha Shinde
S.R.T.M. University Nanded,
Maharashtra India
anushindebiotech@yahoo.com
ABSTRACT
The present invention relates to a filamentous fungus useful for the production of heterologous polypeptides, having been modified by recombinant DNA technology in a manner by which the expression of alkaline proteases have been completely or partially inactivated. The invention also encompasses processes for the production of proteins of interest in high yields by using the fungi of the invention. The invention furthermore relates to methods for producing such fungi and DNA constructs to be used in these methods. Extracellular alkaline proteases from Neurospora crassa is produced for the commercial requirement of proteases. The purification procedure consisted of an ammonium sulfate precipitation, dialysis, and anion-exchange chromatography, and gel filtration. We found only one enzyme identical by polyacrylamide gel electrophoresis. Optimization was done with characterization of enzyme. With a pH value of 10.0 and temperature 60 c the enzyme production was increased.