Extraction and Characterization of Cellulase Enzyme from Teredinidae Marine Wood Borer

Nur Dalila, Mohd Nazri (2018) Extraction and Characterization of Cellulase Enzyme from Teredinidae Marine Wood Borer. [Final Year Project Report] (Unpublished)

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Abstract

Teredinidae or called as shipworms is one of the biological diversity that live in the marine environment. This mollusks enters submerged timbers when it is very small and grows rapidly inside the wood. The shipworms utilize wood as their nutritional source. This wood fragments scraped from the anterior end of the tunnel and enter digestive tract. The activity of the cellulase enzyme from the Teredinidae need to be determined in understanding the digestive activity of this marine wood borer. This study were initiated to extract the cellulase enzyme from the intact tissue of the Teredinidae and characterized their cellulase activity in digesting the woods. Besides, the isolation and determining the availability of the cellulolytic bacteria of the Teredinidae marine wood borer were done in better understanding of the mechanism of wood digestion in marine wood borer. An approach by using the CMC agar plate and Congo red staining were carried out in order to isolate the cellulolytic bacteria from the shipworms. Besides, the Bradford Assay and SDS-PAGE analysis showed that the protein extracted from the guts of the shipworms are more concentrated compared to the protein extracted from the gills of the shipworms. Furthermore, three cellulolytic bacteria were successfully isolated from gills of the shipworms and the amplified 16S rRNA gene of those cellulolytic bacteria produced the PCR reaction product of 1500 bp. The sequencing results that were alignment with BLAST- showed that those cellulolytic bacteria were Streptomyces sp., Azospirillium sp. and Cellulosimicrobium sp. The enzymes extracted from the Streptomyces sp., gills and guts of the shipworms also showed the cellulase activity when tested with Filter Paper Activity and Native-PAGE. The molecular weight of the cellulase extracted from both gills and guts of shipworms were determined approximately 48 kDa by using non-denaturing SDS-PAGE.

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