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On-column refolding of recombinant fungal endoglucanase

Jamaluddin, Mohd Jamil Aizat and Mohd. Salleh, Hamzah (2012) On-column refolding of recombinant fungal endoglucanase. Australian Journal of Basic and Applied Sciences, 6 (1). pp. 128-133. ISSN 1991-8178

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Abstract

Endoglucanase is an industrially important enzyme involved synergistically in degradation of cellulosic materials into constitutive monomers with other cellulase enzymes. In this study, overexpression of (His)6-tagged endoglucanase from Fusarium oxysporum sp. in E. coli BL21(DE3) resulted to high tendency of endoglucanase in the form of inactive inclusion bodies when the host cells were cultured between at 37°C and induced by 0.20 mM (final concentration) IPTG. A simple and time-efficient on-column refolding scheme using immobilized metal-chelate affinity chromatography (IMAC) aided by AKTA purifier, an automated fast protein liquid chromatography (FPLC) system afforded highly purified refolded recombinant endoglucanase 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.

Item Type: Article (Journal)
Additional Information: 1813/23411
Uncontrolled Keywords: On-column refolding; recombinant fungal endoglucanase; inclusion bodies; FPLC
Subjects: T Technology > TA Engineering (General). Civil engineering (General) > TA164 Bioengineering
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: 15 Jun 2012 10:12
Last Modified: 15 Jun 2012 10:12
URI: http://irep.iium.edu.my/id/eprint/23411

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