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KMID : 0545120010110010092
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Journal of Microbiology and Biotechnology
2001 Volume.11 No. 1 p.92 ~ p.96
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Effects of Environmental Factors on In Vivo Folding of Bacillus macerans Cyclodextrin Glycosyltransferase in Recombinant Escherichia coli
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JIN, HEE-HYUN
HAN, NAM SOO/KWEON, DAE-HYUK/PARK, YONG-CHEOL/SEO, JIN-HO
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Abstract
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Effects 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 fraction of the enzyme was increased by lowering the culture temperature to 30℃ and medium pH to 5.8 compared to the enzyme production in LB medium at 37℃ and pH 7.0. Addition of 0.2M NaCl enhanced enzyme expression levels at the expense of cell growth. Glycine betaine that was added after 3h of induction protected not only the cell growth from high osmotic pressure but also helped in vivo folding of CGTase in recombinant E. coli. Addition of 1mM CaCl_2 was also effective in the expression of soluble CGTase, resulting in 15U/ml of the enzyme activity.
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