Analysis of Extracellularly Expressed Recombinant Chitinase by Pichia pastoris.

Aida Syaleeza, Sha'udin (2018) Analysis of Extracellularly Expressed Recombinant Chitinase by Pichia pastoris. [Final Year Project Report] (Unpublished)

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Chitinase is an enzyme that serves different functions for different organisms. Plant chitinase that has been derived from Metroxylon sagu can act as a catalyst for the chitin degradation at biopolymer of Nacetylglucosamine which important in plant defense mechanism against pathogenic insects and fungi. Besides that, it can convert chitinous waste of marine organisms into simple substance thus can help in reducing sea pollution. Chitinase also can be used as an antifungal agent for various fungal infections. For that reason, production of recombinant chitinase is very useful for biotechnology applications. An optimized condition for Pichia pastoris is vital for the recombinant chitinase production. In this project, the isolation of crude recombinant chitinase from the secreted expression in P. pastoris was tested on five different fungi species which are Aspergillus versicolor, Colletotrichum gloeosporioides, Colletotrichum truncatum, Cunninghamella bainieri and Penicillium pinophilum. Results showed that that the existence of chitinase in the secreted expression cannot be proved based on the SDS-PAGE result, but it is not necessarily absence because it might be due to the low concentration of the protein and it is below the detection limit of Coomassie blue. From the Bradford assay, it shows that there is a possibility the recombinant crude protein does exist but in minimal quantity compared to the native crude protein. Testing of recombinant crude protein on the fungal growth using different concentrations (10X, 15X and 20X) showed no inhibition zone.

Item Type: Final Year Project Report
Additional Information: Project Report (B.Sc.) -- Universiti Malaysia Sarawak, 2018.
Uncontrolled Keywords: Chitinase, recombinant, Pichia pastoris.
Subjects: Q Science > Q Science (General)
Q Science > QR Microbiology
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Unai
Date Deposited: 14 Oct 2021 04:40
Last Modified: 14 Oct 2021 04:40

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