Isolation And Characterisation Of Xylanase Gene From Locally Isolated Xylanolytic Bacteria, Bacillus Licheniformis

Tong, Lai Ling (2014) Isolation And Characterisation Of Xylanase Gene From Locally Isolated Xylanolytic Bacteria, Bacillus Licheniformis. [Final Year Project Report] (Unpublished)

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Abstract

Xylan is polysaccharides made from units of xylose which is the major component of hemicelluloses in plant cell wall. Xylanase is the main enzyme for xylan degradation by breaking down the xylan into xylose. Xylanase can undergo bioconversion of lignocellulosic biomass to xylose and further fermented by yeast or bacteria to produce biological fuels. Bioconversion of agricultural waste is a potential renewable energy source that can reduce pollution. Xylanase produced from Bacillus Licheniformis degrade xylan. Xylanase enzyme production and assay was done on B.Licheniformisand the result showed that highest production of enzyme was on the fifth day with O.077U/mL of xylanaseactivity. Besides, the isolation of xylanase gene from B. Licheniformiswas attempted using PCR. A product of about 642 bp was successfully amplified from this xylanolytic bacterial. The Nucleotide BLAST search for the sequencing result showed that 642 bp PCR product was the family 11-endo-IA-beta-xylanase gene with 99% homology with Bacillus subtilisxylanase. In this study, B.Licheniformiswas used and it showed 96% homology with B. licheniformisxylanase. The PCR product was later cloned into pGEM-T Easy Vector and transformed into E. coli XL-l Blue competent cell. The plasmid was successfully extracted and the insert was confirmed using PCR where the extracted pGEM-T xylanaseplasmid served as the template in this application.

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