Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries

Functional metagenomic approach has proven to be powerful tool for identifying novel biocatalysts through high-throughput screening of DNA metagenomic library. Culture enrichment strategies are additional pre-screening methods to provide an attractive means of enhancing the screening hit rate. In...

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Main Authors:	Ibrahim Ali , Noorbatcha, Hamzah, Mohd. Salleh
Format:	Conference or Workshop Item
Language:	English English English
Published:	2015
Subjects:	TP248.13 Biotechnology
Online Access:	http://irep.iium.edu.my/46339/ http://irep.iium.edu.my/46339/1/ICAST_2015_Enrichment_abstract_ENV-EO9.pdf http://irep.iium.edu.my/46339/3/icast_brochure2015.pdf http://irep.iium.edu.my/46339/10/iCAST_2015_Enrichment.pdf

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recordtype	eprints
spelling	iium-463392016-01-06T07:39:58Z http://irep.iium.edu.my/46339/ Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries Ibrahim Ali , Noorbatcha Hamzah, Mohd. Salleh TP248.13 Biotechnology Functional metagenomic approach has proven to be powerful tool for identifying novel biocatalysts through high-throughput screening of DNA metagenomic library. Culture enrichment strategies are additional pre-screening methods to provide an attractive means of enhancing the screening hit rate. In this work metagenomic DNA library was made from Malaysian palm oil mill effluent (POME) microorganisms after culture enrichment experiment basing on natural selection principle. POME microorganisms were inoculated in a medium with similar chemical composition of POME and under controlled conditions of temperature, light, pH with the presence of carboxylmethylcellulose (CMC) for 10 days in order to allow growth of particular microorganisms with cellulose-degrading activities. Metagenomic DNA was extracted from both enriched and non-enriched cultures, 40kb sequences were end-repaired and migrated through 20cm agarose gel electrophoresis showing enhanced DNA concentration of enriched culture compared to the non-enriched culture. These DNA sequences were cloned to PCC1FOS fosmid, packaged in lambda phage and transformed into EPI300T1R. Titer test has shown that the cloning resulted in a high number of colonies from enriched cultures compared to normal metagenomic library. Further high throughput screening is being carried out to test the efficiency of enrichment strategy by comparing positive clones’ frequency between enriched and non-enriched libraries. Key words: cellulose-degrading enzyme, Malaysian palm oil mill effluent (POME), enrichment, metagenomic DNA library, high-throughput screening. 2015 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/46339/1/ICAST_2015_Enrichment_abstract_ENV-EO9.pdf application/pdf en http://irep.iium.edu.my/46339/3/icast_brochure2015.pdf application/pdf en http://irep.iium.edu.my/46339/10/iCAST_2015_Enrichment.pdf Ibrahim Ali , Noorbatcha and Hamzah, Mohd. Salleh (2015) Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries. In: 5th International Conference on Advancement in Science and Technology (iCAST 2015), 10th-12th August 2015, Impiana Resort Cherating, Kuantan, Pahang. (Unpublished)
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English English
topic	TP248.13 Biotechnology
spellingShingle	TP248.13 Biotechnology Ibrahim Ali , Noorbatcha Hamzah, Mohd. Salleh Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
description	Functional metagenomic approach has proven to be powerful tool for identifying novel biocatalysts through high-throughput screening of DNA metagenomic library. Culture enrichment strategies are additional pre-screening methods to provide an attractive means of enhancing the screening hit rate. In this work metagenomic DNA library was made from Malaysian palm oil mill effluent (POME) microorganisms after culture enrichment experiment basing on natural selection principle. POME microorganisms were inoculated in a medium with similar chemical composition of POME and under controlled conditions of temperature, light, pH with the presence of carboxylmethylcellulose (CMC) for 10 days in order to allow growth of particular microorganisms with cellulose-degrading activities. Metagenomic DNA was extracted from both enriched and non-enriched cultures, 40kb sequences were end-repaired and migrated through 20cm agarose gel electrophoresis showing enhanced DNA concentration of enriched culture compared to the non-enriched culture. These DNA sequences were cloned to PCC1FOS fosmid, packaged in lambda phage and transformed into EPI300T1R. Titer test has shown that the cloning resulted in a high number of colonies from enriched cultures compared to normal metagenomic library. Further high throughput screening is being carried out to test the efficiency of enrichment strategy by comparing positive clones’ frequency between enriched and non-enriched libraries. Key words: cellulose-degrading enzyme, Malaysian palm oil mill effluent (POME), enrichment, metagenomic DNA library, high-throughput screening.
format	Conference or Workshop Item
author	Ibrahim Ali , Noorbatcha Hamzah, Mohd. Salleh
author_facet	Ibrahim Ali , Noorbatcha Hamzah, Mohd. Salleh
author_sort	Ibrahim Ali , Noorbatcha
title	Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
title_short	Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
title_full	Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
title_fullStr	Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
title_full_unstemmed	Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries
title_sort	enrichment strategy to enhance screening for novel cellulose-degrading enzymes from malaysian palm oil mill effluent by constructing functional dna metagenomic libraries
publishDate	2015
url	http://irep.iium.edu.my/46339/ http://irep.iium.edu.my/46339/1/ICAST_2015_Enrichment_abstract_ENV-EO9.pdf http://irep.iium.edu.my/46339/3/icast_brochure2015.pdf http://irep.iium.edu.my/46339/10/iCAST_2015_Enrichment.pdf
first_indexed	2023-09-18T21:05:58Z
last_indexed	2023-09-18T21:05:58Z
_version_	1777410935484317696

Enrichment strategy to enhance screening for novel cellulose-degrading enzymes from Malaysian palm oil mill effluent by constructing functional DNA metagenomic libraries

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