Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology

Co-expression of cyclodextrin glucanotransferase (CGTase) from Bacillus sp. G1 (cgt) with bacteriocin release protein (BRP) in Escherichia coli system resulted in the expression and excretion of the enzyme into the culture medium. The cultivation conditions were then optimized with the objective to...

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Main Authors: Lo, Po Kim, Hassan, Osman, Ahmad, Arshad, Mahadi, Nor Muhammad, Illias, Rosli Md
Format: Article
Language:English
Published: Elsevier Inc. 2007
Subjects:
Online Access:http://eprints.utm.my/7254/
http://eprints.utm.my/7254/1/IlliasRosliMD2007_ExcretoryOver-expressionBacillusSp.GI.pdf
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author Lo, Po Kim
Hassan, Osman
Ahmad, Arshad
Mahadi, Nor Muhammad
Illias, Rosli Md
author_facet Lo, Po Kim
Hassan, Osman
Ahmad, Arshad
Mahadi, Nor Muhammad
Illias, Rosli Md
author_sort Lo, Po Kim
building UTeM Institutional Repository
collection Online Access
description Co-expression of cyclodextrin glucanotransferase (CGTase) from Bacillus sp. G1 (cgt) with bacteriocin release protein (BRP) in Escherichia coli system resulted in the expression and excretion of the enzyme into the culture medium. The cultivation conditions were then optimized with the objective to enhance the production of extracellular recombinant CGTase using response surface methodology (RSM) that based on rotatable central composite design (CCD). The process consisted of a total of 50 experiments involving 10 star points and 8 replicates at the central points. The optimum predicted cultivation conditions for the maximum expression of extracellular recombinant CGTase were found to be comprised of: 20 °C post-induction temperature, induction-starting time when cell optical density is 0.3 at 600 nm, 1.0 mM xylose, 50 μM IPTG and 29 h post-induction time, with a predicted extracellular recombinant CGTase activity of 9144.28 U/ml. The experimental extracellular recombinant CGTase activity obtained was 9542.30 U/ml, which was very close to the predicted value. The expression of extracellular recombinant CGTase improved about 151-fold after the optimization was conducted.
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spelling utm-72542017-10-22T08:53:59Z http://eprints.utm.my/7254/ Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology Lo, Po Kim Hassan, Osman Ahmad, Arshad Mahadi, Nor Muhammad Illias, Rosli Md TP Chemical technology Co-expression of cyclodextrin glucanotransferase (CGTase) from Bacillus sp. G1 (cgt) with bacteriocin release protein (BRP) in Escherichia coli system resulted in the expression and excretion of the enzyme into the culture medium. The cultivation conditions were then optimized with the objective to enhance the production of extracellular recombinant CGTase using response surface methodology (RSM) that based on rotatable central composite design (CCD). The process consisted of a total of 50 experiments involving 10 star points and 8 replicates at the central points. The optimum predicted cultivation conditions for the maximum expression of extracellular recombinant CGTase were found to be comprised of: 20 °C post-induction temperature, induction-starting time when cell optical density is 0.3 at 600 nm, 1.0 mM xylose, 50 μM IPTG and 29 h post-induction time, with a predicted extracellular recombinant CGTase activity of 9144.28 U/ml. The experimental extracellular recombinant CGTase activity obtained was 9542.30 U/ml, which was very close to the predicted value. The expression of extracellular recombinant CGTase improved about 151-fold after the optimization was conducted. Elsevier Inc. 2007-04-03 Article PeerReviewed application/pdf en http://eprints.utm.my/7254/1/IlliasRosliMD2007_ExcretoryOver-expressionBacillusSp.GI.pdf Lo, Po Kim and Hassan, Osman and Ahmad, Arshad and Mahadi, Nor Muhammad and Illias, Rosli Md (2007) Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology. Enzyme and Microbial Technology, 40 (5). pp. 1256-1263. ISSN 0141-0229 http://dx.doi.org/10.1016/j.enzmictec.2006.09.020 10.1016/j.enzmictec.2006.09.020
spellingShingle TP Chemical technology
Lo, Po Kim
Hassan, Osman
Ahmad, Arshad
Mahadi, Nor Muhammad
Illias, Rosli Md
Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title_full Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title_fullStr Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title_full_unstemmed Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title_short Excretory over-expression of Bacillus sp. G1 cyclodextrin glucanotransferase (CGTase) in Escherichia coli: Optimization of the cultivation conditions by response surface methodology
title_sort excretory over-expression of bacillus sp. g1 cyclodextrin glucanotransferase (cgtase) in escherichia coli: optimization of the cultivation conditions by response surface methodology
topic TP Chemical technology
url http://eprints.utm.my/7254/
http://eprints.utm.my/7254/
http://eprints.utm.my/7254/
http://eprints.utm.my/7254/1/IlliasRosliMD2007_ExcretoryOver-expressionBacillusSp.GI.pdf