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On-column refolding of recombinant fungal endoglucanase I

Aizat Jamaluddin, Mohd Jamil and Mohd. Salleh, Hamzah (2011) On-column refolding of recombinant fungal endoglucanase I. In: 2nd International Conference on Biotechnology Engineering (ICBioE 2011), 17-19 May 2011, The Legend Hotel, Kuala Lumpur.

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Abstract

Endoglucanase is an industrially important enzyme involved in degradation of cellulosic materials into constitutive monomers synergistically with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase I from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high tendency of endoglucanase I in the form of inactive inclusion bodies when the host cells were cultured between 25-37°C and induced by 0.05-0.20 mM (final concentration) IPTG. A simple and time-efficient on-column refolding scheme using IMAC aided by AKTA purifier, an automated fast protein liquid chromatography (FPLC) system afforded highly purified refolded recombinant endoglucanase I that was active in carboxymethyl cellulose (CMC) assay. The on-column refolding scheme is reliable and can be applied for efficient and robust method for lab-scale production of recombinant endoglucanase I.

Item Type: Conference or Workshop Item (Full Paper)
Additional Information: 1813/16211
Uncontrolled Keywords: On-column refolding; recombinant fungal endoglucanase I; inclusion bodies; FPLC.
Subjects: T Technology > TP Chemical technology > TP248.13 Biotechnology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button): Kulliyyah of Engineering > Department of Biotechnology Engineering
Depositing User: Dr Hamzah Mohd. Salleh
Date Deposited: 02 Feb 2012 12:15
Last Modified: 02 Feb 2012 15:16
URI: http://irep.iium.edu.my/id/eprint/16211

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