Identification and enzyme production of a cellulolytic Bacillus-strain isolated from moose (Alces alces) rumen

Mastergradsoppgave i næringsrettet bioteknologi, Avdeling for lærerutdanning og naturvitenskap, Høgskolen i Hedmark, 2013. Master of applied and commercial biotechnology. A major obstacle to industrial-scale production of fuel from lignocellulose lies in the inefficient deconstruction of plant mater...

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Bibliographic Details
Main Author: Sanusi, Oluwaseyi Adelaja
Format: Master Thesis
Language:English
Published: 2013
Subjects:
Online Access:http://hdl.handle.net/11250/278760
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Summary:Mastergradsoppgave i næringsrettet bioteknologi, Avdeling for lærerutdanning og naturvitenskap, Høgskolen i Hedmark, 2013. Master of applied and commercial biotechnology. A major obstacle to industrial-scale production of fuel from lignocellulose lies in the inefficient deconstruction of plant material, due to the recalcitrant nature of the substrate toward enzymatic breakdown and the relatively low activity of currently available hydrolytic enzymes. Improvement of the process of cellulase production and development of more efficient lignocellulose-degrading enzymes are necessary in order to reduce the cost of enzymes required in the biomass-to-bioethanol process. Cellulases are required for cellulose degradation in nature and almost all of the biomass produced is mineralized again by enzymes which are provided by microorganisms. The crystalline material is hydrolyzed by a number of simultaneously present, interacting enzymes (endoglucanase, exoclucanase and β-glucanase), or alternatively by a multienzyme complex. Cellulosome complexes are intricate multi-enzyme machines produced by many cellulolytic microorganisms. They are characterized by having a scaffolding protein, and are typically anchored to the cell membrane through a dockerin-protein. The goal of this work involves the production, identification and initial purification of a cellulolytic and hemicellulolytic enzymes from bacterial isolates from moose (Alces alces) rumen. Five bacterial isolates (MRB 1-5) were comparatively analysed for effective producer of cellulase enzyme. Isolates were screened for cellulolytic activity using Carboxy Methyl Cellulose (CMC) agar plates and DNS reducing sugar assay, these techniques are time-efficient and reliable in identification of cellulolytic microorganisms. Screening also included growth curve characteristics under anaerobic and aerobic conditions. Among the five bacterial isolates, isolate MRB 3 was found to be the most effective cellulase producer both qualitatively and quantitatively. MRB 3 was identified ...