Isolation and identification of cellulolytic bacteria from the larval gut of rhynchophorus ferrugineus / Nurul Hidayah binti Adenan

Adenan, Nurul Hidayah (2013) Isolation and identification of cellulolytic bacteria from the larval gut of rhynchophorus ferrugineus / Nurul Hidayah binti Adenan. Masters thesis, University of Malaya.

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Abstract

This project is carried out to investigate the profiling of unculturable and culturable cellulolytic bacteria from larval guts of Rhynchophorus ferruginase which had been fed with different type of lignocelullosic materials for two weeks. The physicochemical analysis of baggase, oil palm trunk, and sago were processed using AOAC method. The result in percentage dry weight samples indicated that oil palm trunk contained 39.37% of cellulose and 18.53% of hemicelluloses while baggase 37.04% and 23.8%, respectively. The lower amount of cellulose and hemicelluloses were observed in sago trunk due to sample was taken after eaten by the larvae. The gut of Rhynchophorus ferrugineus were proceeds by dissecting and dividing all guts into three major sections known as foregut, midgut and hindgut. Each section was homogenized and used for further analysis. Isolation of cellulolytic bacteria was performed using TSA media. Screening of cellulolytic bacteria was carried out by using CMC plate agar and Congo red stain dye. Strain 18 and 11 indicated the highest enzyme activity with 18 and 15mm of holozone diameter. Results on enzyme assay revealed that maximum yield of FPase, CMCase and β-glucosidase are 0.48 and 0.46 U/mL, 0.31 and 0.47 U/mL, 0.18 and 0.26 U/mL, respectively.Their carbon preferences (N-Acetyl-D-Glucosamine, D-Glucose-6-PO4, D-Raffinose, D-Fructose-6-PO4) were analysed by using biolog to further confirmed the result from selected strain. The study of unculturable cellulolytic bacterial community was done using molecular analysis which is Polymerase Chain Reaction and Denaturing Gradient Gel Electrophoresis. As a result, two cellulolytic bacterial (Alcaligenes spp and Enterobacter aerogenes) were detected with the species identification by using Basic Local Alignment Search Tool (Blastn) and CLUSTALW.

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