Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking

hive of the best endoglucanase producers has been successfully isolated and identified based on the 'halo' formed on Luria Bertani (LB) agar media containing 1% (w/v) of carboxymethylcellulose (CMC) flooded with Congo Red solution. Molecular approach targeting the conserved region of 16S...

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Main Author:	Hashimatul Fatma, Hashim
Format:	Thesis
Language:	English
Published:	2008
Subjects:	Q Science (General) QP Physiology
Online Access:	http://ir.unimas.my/id/eprint/28817/1/Hashimatul.pdf
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id	my-unimas-ir.28817
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spelling	my-unimas-ir.288172023-05-08T07:46:44Z Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking 2008 Hashimatul Fatma, Hashim Q Science (General) QP Physiology hive of the best endoglucanase producers has been successfully isolated and identified based on the 'halo' formed on Luria Bertani (LB) agar media containing 1% (w/v) of carboxymethylcellulose (CMC) flooded with Congo Red solution. Molecular approach targeting the conserved region of 16S rRNA bacterial strains was used to further confirm the initial morphological and biochemical tests result. A phylogenetic tree construction using Neighbour-Joining, Maximum Parsimony and Maximum Likelihood analysis was used to further strengthen the findin he best endoglucanase producer strain in this research was found to be bacterial isolate of Bacillus licheniformis BL-P7. PCR amplification using endoglucanase primers produced a PCR product of 750 bp in molecular weight of size. In silico analysis of sequencing product indicate this gene is coding for an extracellular alkaline endo-I, 4-beta-glucanase. In this study, partial of endoglucanase gene has been successfully was cloned and characterized however no expression was obtained. Bacillus licheniformis BL-P7 was inoculated into submerged liquid fermentation media utilizing sago pith waste and rice husk. An optimal condition for endoglucanase secretion by Bacillus licheniformis BL-P7 was detennined successfully. Preliminary kinetics study on crude endoglucanase enzyme was stable in wide spectrum of pH and at high temperature. Purification of the crude enzyme extract of Bacillus licheniformis BL-P7 produced in the submerged liquid fermentation to apparent homogeneity was carried out. The isoelectric point (pI) for the purified enzyme was 5.5 with the protein weight size of 35 kDa. The purified endoglucanase was very stable in wide range of pH (up to pH 12) and temperature (up to 100 0C). The enzyme action is unaffected in the presence of various metal ions. This research has conducted the trial enzymatic deinking on mixed office paper. The results revealed that enzymatic de inking profoundly have better results compared to conventional chemical deinking. Removal of ink residues and all paper properties were enhanced in terms of brightness, air permeable, tensile and tear with 50% efficiency. Universiti Malaysia Sarawak (UNIMAS) 2008 Thesis http://ir.unimas.my/id/eprint/28817/ http://ir.unimas.my/id/eprint/28817/1/Hashimatul.pdf text en validuser masters Universiti Malaysia Sarawak (UNIMAS) Faculty of Resource Science and Technology
institution	Universiti Malaysia Sarawak
collection	UNIMAS Institutional Repository
language	English
topic	Q Science (General) QP Physiology
spellingShingle	Q Science (General) QP Physiology Hashimatul Fatma, Hashim Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
description	hive of the best endoglucanase producers has been successfully isolated and identified based on the 'halo' formed on Luria Bertani (LB) agar media containing 1% (w/v) of carboxymethylcellulose (CMC) flooded with Congo Red solution. Molecular approach targeting the conserved region of 16S rRNA bacterial strains was used to further confirm the initial morphological and biochemical tests result. A phylogenetic tree construction using Neighbour-Joining, Maximum Parsimony and Maximum Likelihood analysis was used to further strengthen the findin he best endoglucanase producer strain in this research was found to be bacterial isolate of Bacillus licheniformis BL-P7. PCR amplification using endoglucanase primers produced a PCR product of 750 bp in molecular weight of size. In silico analysis of sequencing product indicate this gene is coding for an extracellular alkaline endo-I, 4-beta-glucanase. In this study, partial of endoglucanase gene has been successfully was cloned and characterized however no expression was obtained. Bacillus licheniformis BL-P7 was inoculated into submerged liquid fermentation media utilizing sago pith waste and rice husk. An optimal condition for endoglucanase secretion by Bacillus licheniformis BL-P7 was detennined successfully. Preliminary kinetics study on crude endoglucanase enzyme was stable in wide spectrum of pH and at high temperature. Purification of the crude enzyme extract of Bacillus licheniformis BL-P7 produced in the submerged liquid fermentation to apparent homogeneity was carried out. The isoelectric point (pI) for the purified enzyme was 5.5 with the protein weight size of 35 kDa. The purified endoglucanase was very stable in wide range of pH (up to pH 12) and temperature (up to 100 0C). The enzyme action is unaffected in the presence of various metal ions. This research has conducted the trial enzymatic deinking on mixed office paper. The results revealed that enzymatic de inking profoundly have better results compared to conventional chemical deinking. Removal of ink residues and all paper properties were enhanced in terms of brightness, air permeable, tensile and tear with 50% efficiency.
format	Thesis
qualification_level	Master's degree
author	Hashimatul Fatma, Hashim
author_facet	Hashimatul Fatma, Hashim
author_sort	Hashimatul Fatma, Hashim
title	Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
title_short	Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
title_full	Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
title_fullStr	Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
title_full_unstemmed	Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
title_sort	identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking
granting_institution	Universiti Malaysia Sarawak (UNIMAS)
granting_department	Faculty of Resource Science and Technology
publishDate	2008
url	http://ir.unimas.my/id/eprint/28817/1/Hashimatul.pdf
_version_	1783728363699961856

Identification and characterizationof novel cellulase from thermophilic microbial consortium for potential application in enzymatic deinking

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