TITLE:
Characterization of Exo 1, 4-β glucanase produced from Tricoderma Viridi MBL through solid-state bio-processing of orange peel waste
AUTHORS:
Muhammad Irshad, Zahid Anwar, Amber Afroz
KEYWORDS:
Orange Peel Waste; Exo 1, 4-β Glucanase; T. Viride MBL; Purification; SDS-PAGE
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.3 No.5,
September
21,
2012
ABSTRACT: Agro-industrial residues are primarily composed of complex polysaccharides that strengthen the microbial growth for the production of industrially important enzymes like cellulases. In the present study we aimed to characterize the Exo 1, 4-β glucanase that was indigenously produced from Trichoderma viride MBL. T. viride MBL was cultured in the Solid-State medium of orange peel (50% w/w moisture) under optimized fermentation conditions and maximum activity of 412 ± 12 U/mL was recorded after 4th day of incubation at pH 5.5 and 30℃. Exo 1, 4-β glucanase was 4.17-fold purified with specific activity of 642 U/mg in comparison to the crude extract. To confirm its purity and molecular weight, sodium dodecyl sulphate poly acrylamide gel electrophoresis (SDS- PAGE) was performed. The enzyme was shown to have a molecular weight of 60 kDa with an optimum pH and temperature of 5 and 50℃, respectively. Lineweaver-Burk reciprocal plot revealed that the kinetic constants Km and Vmax of purified Exo 1, 4-β glucanase were 76 μM and 240 U/mL.