CHARACTERIZATION OF A NOVEL MANGANESE PEROXIDASE PURIFIED FROM SOLID STATE CULTURE OF Trametes versicolor IBL-04

Muhammad Asgher, Hafiz MN Iqbal

Abstract


A novel manganese peroxidase (MnP) produced by an indigenous white rot fungal strain Trametes versicolor IBL-04 in solid state medium of corncobs was purified and characterized. The fungus produced 964U/mL MnP in the presence of additional carbon (glucose) and nitrogen (yeast extract) supplements added at a C:N ratio of 25:1, 1mM Tween-80 (1mL), 1mM MnSO4 (1mL), and 1mM CuSO4 (1mL). The MnP was purified by ammonium sulfate fractionation (65% saturation) and dialysis, followed by Sephadex G-100 gel filtration chromatography. Purification procedures resulted in 2.4-fold purification with an overall yield and specific activity of 3.4% and 660 U/mg, respectively. The purified MnP was monomeric of molecular weight of 43 kDa, showing a single band on sodium dodecyl sulfate poly acrylamide gel electrophoresis (SDS-PAGE). The enzyme was optimally active at pH 5 and 50oC and was stable for 1 h over a broad range of pH (4-7) and temperature (40-65oC). Kinetic constants KM and Vmax of purified MnP were 70 µM and 540 U/mL for MnSO4 substrate. The effect of possible activators and inhibitors of enzyme were also investigated, and it was observed that EDTA, Cystein, and Ag+ caused MnP inhibition and inactivation to different extents, whereas MnP was activated by 4 and 3 mM of Cu2+ and Fe2+, respectively. High thermo-stability, low KM and high Vmax features of this novel MnP isolated from culture filtrate of T. versicolor IBL-04 suggests its suitability for various industrial and biotechnological applications.

Keywords


T. versicolor IBL-04; Manganese peroxidase; Purification; SDS-PAGE; Characterization

Full Text: PDF

Welcome to BioResources! This online, peer-reviewed journal is devoted to the science and engineering of biomaterials and chemicals from lignocellulosic sources for new end uses and new capabilities. The editors of BioResources would be very happy to assist you during the process of submitting or reviewing articles. Please note that logging in is required in order to submit or review articles. Martin A. Hubbe, (919) 513-3022, hubbe@ncsu.edu; Lucian A. Lucia, (919) 515-7707, lucian.lucia@gmail.com URLs: bioresourcesjournal.com; http://ncsu.edu/bioresources ISSN: 1930-2126