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Simultaneous refolding, purification, and immobilization of recombinant Fibrobacter succinogenes 1,3-1,4-β-D-glucanase on artificial oil bodies
- Source :
- Journal of Chemical Technology & Biotechnology. 84:1480-1485
- Publication Year :
- 2009
- Publisher :
- Wiley, 2009.
-
Abstract
- BACKGROUND: 1,3-1,4-β-D-glucanase (1,3-1,4-β-D-glucan 4-glucanohydrolase; EC 3.2.1.73) has been used in a range of industrial processes. As a biocatalyst, it is better to use immobilized enzymes than free enzymes, therefore, the immobilization of 1,3-1,4-β-D-glucanase was investigated. RESULTS: A 1,3-1,4-β-D-glucanase gene from Fibrobacter succinogenes was overexpressed in Escherichia coli as a recombinant protein fused to the N terminus of oleosin, a unique structural protein of seed oil bodies. With the reconstitution of the artificial oil bodies (AOBs), refolding, purification, and immobilization of active 1,3-1,4-β-D-glucanase was accomplished simultaneously. Response surface modeling (RSM), with central composite design (CCD), and regression analysis were successfully applied to determine the optimal temperature and pH conditions of the AOB-immobilized 1,3-1,4-β-D-glucanase. The optimal conditions for the highest immobilized 1,3-1,4-β-D-glucanase activity (7.1 IU mg−1 of total protein) were observed at 39 °C and pH 8.8. Furthermore, AOB-immobilized 1,3-1,4-β-D-glucanase retained more than 70% of its initial activity after 120 min at 39 °C, and it was easily and simply recovered from the surface of the solution by brief centrifugation; it could be reused eight times while retaining more than 80% of its activity. CONCLUSIONS: These results indicate that the AOB-based system is a comparatively simple and effective method for simultaneous refolding, purification, and immobilization of 1,3-1,4-β-D-glucanase. Copyright © 2009 Society of Chemical Industry
- Subjects :
- Fibrobacter succinogenes
Central composite design
Immobilized enzyme
biology
Renewable Energy, Sustainability and the Environment
General Chemical Engineering
Organic Chemistry
Cellulase
medicine.disease_cause
Pollution
Inorganic Chemistry
Fuel Technology
Biochemistry
medicine
Xylanase
biology.protein
Centrifugation
Oleosin
Waste Management and Disposal
Escherichia coli
Biotechnology
Subjects
Details
- ISSN :
- 02682575
- Volume :
- 84
- Database :
- OpenAIRE
- Journal :
- Journal of Chemical Technology & Biotechnology
- Accession number :
- edsair.doi...........f0e6bfc6c3c24e245f6619d80d36cd5c
- Full Text :
- https://doi.org/10.1002/jctb.2205