Isolation and characterization of a thermostable cellulase from thermophilic microorganisms obtained Anah Rais hot spring

Noor Izzati, Yacob (2005) Isolation and characterization of a thermostable cellulase from thermophilic microorganisms obtained Anah Rais hot spring. [Final Year Project Report] (Unpublished)

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Abstract

this study concerned about the isolation and identification of thermophilic microorganisms that has the thermostable enzyme obtained frum Anah Rais Hot Spring. On minimal media containing CMC, five colonies of microorganisms have been isolated and identified as AR1 Bacillus spp., AR2 Nisseria spp., AR3 Enterobacter spp., AK4 Streptococcus spp., and AR5 Klebsiella spp. Cellulase enzyme activity was indicated by enzyme degradation of the CMC media that produced clear haloes around the supernatant (containing the enzyme) of each microorganism after flooded with Congo red. Later, the microorganisms were grown on fermentation media consists of O.5% w/v sago and paddy pith waste. 0.7% w/v yeast extract, 0.1% w/v KH2PO4 and 0.5% w/v MgSO. The enzyme activity was determined using DNS and FPase enzyme assay. Bradford assay was used to determine the protein content. Isolate show the highest cellulolytic activity was preceded to enzyme purification that comprised ((NH4)2SO4) precipitation and HIC. SDS-PAGE was used to determine the purity and molecular weight of cellulase enzyme. Genomic DNA from each microorganism were extracted and used for molecular identification. Through enzyme assay, isolate of AR4 Streptococcus spp was characterized as the best thermophilic microorganisms. The crude enzymec of the isolate was found to show highest enzyme activity (Sago/paddy pith waste media of 0.428U/ml) within 15 hour at temperature 55°C during enzyme substrate reaction. However, gene encoding cellulase enzyme from this isolate could not obtain.

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