Decolorization and purification of crude protease from Rhizopus oryzae by activated charcoal and its electrophoretic analysis

Citation
K. Aikat et al., Decolorization and purification of crude protease from Rhizopus oryzae by activated charcoal and its electrophoretic analysis, BIOTECH LET, 23(4), 2001, pp. 295-301
Citations number
23
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY LETTERS
ISSN journal
01415492 → ACNP
Volume
23
Issue
4
Year of publication
2001
Pages
295 - 301
Database
ISI
SICI code
0141-5492(200102)23:4<295:DAPOCP>2.0.ZU;2-I
Abstract
Activated charcoal decolorized and partially purified the protease from a c rude extract of solid state fermentation of wheat bran by Rhizopus oryzae. Treatment for 5 min was sufficient. Depending on the initial colour intensi ty of crude, the charcoal to crude extract ratio could be optimized to achi eve 90% decolorization, 85% enzyme recovery, and over a 3-fold purification , even up to 20-fold variation in batch size (from 1 ml to 20 ml crude extr act). Decolorization followed the Freundlich and the Langmuir models, the F reundlich constant, n, being 2.74. Partial purification was confirmed by na tive PAGE and the protease band identified by gelatin-PAGE. SDS-PAGE showed the protease consisted of two sub-units (about 22 and 24 kDa). List of symbols: c(o), initial solute concentration in liquid before adsorp tion; c*, equilibrium solute concentration in liquid after adsorption; k, e mpirical constant for Freundlich adsorption isotherm; U, unit of protease a ctivity; v, volume of solution per unit weight of adsorbent.