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KMID : 0545120040140010216
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 1 p.216 ~ p.219
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GroEL/ES Chaperone and Low Culture Temperature Synergistically Enhanced the Soluble Expression of CGTase in E. coli
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PARK, SO LIM
KWON, MI JUNG/KIM, SUNG KOO/NAM, SOO WAN
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Abstract
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The effect of culture temperature on the production of soluble form of B. macerans cyclodextrin glucanotransferase (CGTase) in recombinant E. coli was investigated E. coli cell was cotransformed with two plasmids (pTCGTI and pGrol1) in which the cgt and groEL/ES genes are under the control of T7 promoter and pzt-1 promoter, respectively. When tetracycline (10 ng/ml) and IPTG (1 mM) were added as inducers at the early-exponential phase (2 h) and mid-exponential phase (3 h), respectively, the solubilization of the inclusion body CGTase was greatly dependent on the temperature of the culture. At low culture temperature of ZYC, 2 or 3-fold higher activity and specific activity were obtained over 37℃. SDS-PAGE analysis revealed that about 62% of CGTase in the total CGTase protein was found in the soluble fraction by applying overexpression of GroEL/ ES chaperone and by cultivation of E. coli at 25℃, whereas 33% of CGTase was detected in the soluble fraction at 37℃. Therefore, the expression of GroELES and cultivation at 25℃ greatly enhanced the soluble production of CGTase in E. coli.
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KEYWORD
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