Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Effect of Environmental Factors on In Vivo Folding of Bacillus macerans Cyclodextrin Glycosyltransferase in Recombinant Escherichia coli

  • Jin, Hee-Hyun (Department of Food Science and Technology, and Research Center for New-Biomaterials in Agriculture, Seoul National University) ;
  • Han, Nam-Soo (Department of Food Science and Technology, Chonbuk National University) ;
  • Kweon, Dae-Hyuk (Department of Food Science and Technology, and Research Center for New-Biomaterials in Agriculture, Seoul National University) ;
  • Park, Yong-Cheol (Interdisciplinary Program for Biochemical Engineering and Biotechnology, Seoul National University) ;
  • Seo, Jin-Ho (Department of Food Science and Technology, and Research Center for New-Biomaterials in Agriculture, Seoul National University)
  • Published : 2001.02.01
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Abstract

Effect of environmental factors on the expression of soluble forms of Bacillus macerans cyclodextrin glycosyltransferase in recombinant Escherichia coli BL21(DE3)pLysE:pTCGT1 were investigated. The amount of soluble CGTase produced in the cell was measured by determining its enzymatic activity. The soluble fractionof the enzyme was increased by lowering the culture temperature to $30{\circ}C$ and medium pH to 5.8 compared to the enzyme production in LB medium at $37^{\circ}C$ and pH7.0. Addition of 0.2 M NaCl enhanced enzyme expression levels at the expense of cell growth. Glycine betaine that was added after 3 h of induction protected not only the cell growth from hig osmotic pressue but also hepld in vivo folding of CGTase in recombinant E. coli. Addition of 1 mM $CaCl_2$ was also effective in the expression of soluble CGTase, resulting in 15 U/ml of the enzyme activity.

Keywords

References

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