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

Po Kim Lo, Osman Hassan, Arshad Ahmad, Nor Muhammad Mahadi, Rosli Md Illias

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1256-1263
Number of pages8
JournalEnzyme and Microbial Technology
Volume40
Issue number5
DOIs
Publication statusPublished - 3 Apr 2007

Fingerprint

Cyclodextrins
Bacilli
Escherichia coli
Bacillus
Isopropyl Thiogalactoside
Bacteriocins
Xylose
Density (optical)
Escherichia coli Proteins
Stars
Culture Media
cyclomaltodextrin glucanotransferase
Enzymes
Cell Count
Proteins
Temperature
Composite materials
Experiments

Keywords

  • E. coli
  • Extracellular
  • Optimization
  • Recombinant CGTase
  • RSM

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

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.

In: Enzyme and Microbial Technology, Vol. 40, No. 5, 03.04.2007, p. 1256-1263.

Research output: Contribution to journalArticle

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