Cellulase enzyme from palm oil mill effluent: production, characterization and application /
The study was conducted for the production of cellulase enzyme through liquid state bioconversion of palm oil mill effluent (POME) by filamentous fungi and characterization of the produced enzyme. Several experiments such as selection of filamentous fungi, optimization of media and process parameter...
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| Main Author: | |
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| Format: | Thesis |
| Language: | English |
| Published: |
Gombak, Selangor :
Kulliyyah of Engineering, International Islamic University Malaysia,
2011
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| Subjects: | |
| Online Access: | http://studentrepo.iium.edu.my/handle/123456789/4392 |
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| Summary: | The study was conducted for the production of cellulase enzyme through liquid state bioconversion of palm oil mill effluent (POME) by filamentous fungi and characterization of the produced enzyme. Several experiments such as selection of filamentous fungi, optimization of media and process parameters, development of the separation/purification conditions for ultra-filtration, characterization and evaluation of purified cellulase enzyme through hydrolysis of oil palm empty fruit bunches (EFB) were carried out to achieve the entire objectives of the study. The results on isolation indicated that among the strains isolated, the strains IBPC108R, IBEC302B and IBEC305B gave the highest FPAse activity (0.3 U/ml) after 2 days of fermentation and IBEC302B showed the highest CMCase activity (0.42 U/ml) after 3 days of fermentation. Among the laboratory stocks, Aspergillus IBO-102MNB, gave the highest FPAse activity (0.28 U/ml) and Aspergillus IBO-103MNB gave the highest CMCase activity (0.62 U/ml) after 2 days of fermentation. Among the strains tested, the Trichoderma reesei RUT C-30, a commercial strain collected from ATTC, gave highest activity in both FPA and CMC assay after 5 days of fermentation with 0.996 U/ml (FPAse) and 2.89 U/ml (CMCase) respectively which was selected as the potential strain for further development of cellulases production. The optimization of the constituents of the bioconversion medium was conducted sequentially through placket-burman design, one factor at a time (OFAT) method and central composite design (CCD) to increase the production of cellulase with highest activity in a nutrient supplemented palm oil mill effluent (POME). The optimum medium constituents were 0.5% of cellulose, 0.5% of peptone and 0.2% of tween 80 to produce highest activity of cellulase (18.1 CMCase, U/ml). The highest cellulase activity (28.29 CMCase, U/ml) was achieved with agitation of 200 rpm, pH of 7 and aeration of 1.5 vvm as the optimized process conditions through the OFAT and statistical methods. It was observed from the study that the enzyme productivity (as activity) was increased by 6.3 fold and finally 9.8 fold after medium and process optimization respectively. Specific growth rate (μ), maximum specific rate of product formation and the activation energy was estimated to be 0.79 day-1, 3.68 day-1 after second day of bioconversion and 22.68 kJ/mol respectively. The molecular weight of the purified cellulase enzyme was found to be 66 KDa after carrying out the fractionation of the crude enzyme by FPLC and SDS-PAGE. The crude enzyme was purified and concentrated by 9 fold with protein retention 34.67%, permeation 43.89% and fouling 21.44 % using 10 KDa membrane under 1.025 (bar) of TMP during cross-flow micro/ultra-filtration. 30 key parameters of the raw POME and ultra-filtration permeate were examined to evaluate the quality of the waste of the bioconversion process. The highest yield of reducing sugar, 0.54 g/g of EFB was achieved by enzymatic hydrolysis with 4% (w/v) of pretreated EFB, 10% (v/v) of enzyme with activity 60 CMCase (U/ml) after 120 hours of incubation by conducting the statistical method during optimization of the enzymatic saccharification of EFB. The findings of the current study might contribute to the development of the Malaysian economy by showing an alternative economic solution of POME and EFB management through bio-product (cellulase) recovery and production of reducing sugar for bioethanol industry. |
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| Item Description: | "A thesis submitted in fulfilment of the requirement for the degree of Doctor of Philosophy in Engineering (Biotechnology)."--On t.p. |
| Physical Description: | xxiii, 292 leaves : ill. charts ; 30cm. |
| Bibliography: | Includes bibliographical references (leaves 238-272). |