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KMID : 0545120020120050753
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Journal of Microbiology and Biotechnology
2002 Volume.12 No. 5 p.753 ~ p.759
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Extracellular Overproduction of β-Cyclodextrin Glucanotransferase in a Recombinant E. coli Using Secretive Expression System
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LEE, KWANG-WOO
SHIN, HYUN-DONG/LEE, YONG-HYUN
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Abstract
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β-Cyclodextrin glucanotransferase (β-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The β-CGTase gene of alkalophilic Bacillus firmus var. alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL21(DE3)pLysS. The optimum culture cinditions for the overproduction of β-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of 25℃. A significant amount of β-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of β-CGTase ant a high cell density, resulting in production of up to 21.6/Uml of β-CGTase.
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