Performance Improvement of Ca-Alginate Bead Cross- Linked Laccase from Trametes versicolor IBL-04

Sadia Noreen, Muhammad Asgher, Fatma Hussain, Ahsan Iqbal

Abstract


Extracellular laccase was produced by Trametes versicolor IBL-04 using corn cobs as a substrate under pre-optimized culture conditions. A 64-kDa laccase enzyme was purified and immobilized on calcium alginate beads using glutaraldehyde as a cross-linking reagent. Maximum enzyme immobilization efficiency (89%) was observed with 2-mm calcium-alginate beads that were developed using 4% (w/v) sodium alginate in 2% (w/v) calcium chloride solution. Immobilization of laccase enhanced the optimum temperature but caused an acidic shift in the optimum pH of the enzyme. The immobilized enzyme showed optimum activity at pH 3.0 and 60 °C as compared to pH 4.5 and 45 °C for free laccase. The kinetic constants Km and Vmax of laccase were significantly altered by immobilization. The affinity of enzyme toward its substrate increased (Km decreased), leading to enhanced catalytic efficiency (Vmax increased). Scanning electron microscopy (SEM) was performed to characterize the free and enzyme-bound immobilization matrix. Free and immobilized enzymes also were used for decolorization of the Reactive T Blue dye (030905 GWF) for three days. The free and immobilized laccases decolorized the dye by 65% and 92%, respectively, in 72 h. The immobilized enzyme retained 68% of its original activity after three cycles of repeated reuse for dye decolorization, indicating the usefulness of immobilized laccase in repeated industrial batch operations.

Keywords


Laccase; Ca-alginate beads; Immobilization; Thermo-stability; Dye decolorization

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