Biodegradation of 2-chlorophenol using free and immobilized laccase from Trametes Versicolor / Swapna Sambrani
Laccase from white rot fungus Trametes versicolor was immobilized in sodium alginate beads using entrapment method. Kinetic parameters (Vmax and Km) of both free and immobilized laccase were determined using syringaldazine as substrate. 2- chlorophenol was degraded using free and immobilized lacc...
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| Format: | Thesis |
| Published: |
2009
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| Subjects: | |
| Online Access: | http://studentsrepo.um.edu.my/6225/1/Final_Thesis_Report.pdf http://studentsrepo.um.edu.my/6225/2/front_page%2Dmy_thesis.pdf http://studentsrepo.um.edu.my/6225/ |
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| Summary: | Laccase from white rot fungus Trametes versicolor was immobilized in
sodium alginate beads using entrapment method. Kinetic parameters (Vmax and Km) of
both free and immobilized laccase were determined using syringaldazine as substrate. 2-
chlorophenol was degraded using free and immobilized laccase. An indirect rapid assay
method was developed for the assay of degraded products of 2-chlorophenol, wherein the
anionic phenoxy radicals, possibly generated during 2-chlorophenol degradation, couples
with a cationic dye, Methylene Blue and renders it colorless. The extent of decolorization
indicated the extent of product formation and hence the extent of 2-chlorophenol
degradation assuming a 1:1 stoichiometric reaction between degraded products of 2-
chlorophenol and the dye. Factors like pH, temperature and enzyme concentration were
optimized for the free enzyme reaction and reaction time, temperature and enzyme
concentration were optimized for the immobilized enzyme reaction using a statistical tool
viz. Box-Behnken method of Response Surface Methodology. The efficiency of
immobilized laccase with respect to 2-chlorophenol degradation was found to be
comparable to that of free laccase. External mass transfer was found to have negligible
effect on the diffusion of substrate from bulk liquid to the surface of the bead while there
was significant limitation posed by internal mass transfer from the surface of the bead to
the center of the bead. |
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