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KMID : 0545120120220040510
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Journal of Microbiology and Biotechnology
2012 Volume.22 No. 4 p.510 ~ p.515
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Large Increase in Leuconostoc citreum KM20 Dextransucrase Activity Achieved by Changing the Strain/Inducer Combination in an E. coli Expression System
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Ko Jin-A
Jeong Hyung-Jae
Ryu Young-Bae
Park Su-Jin
Wee Young-Jung
Kim Do-Man
Kim Young-Min
Lee Woo-Song
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Abstract
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A recombinant putative dextransucrase (DexT) was produced from Leuconostoc citreum KM20 as a 160 kDa protein, but its productivity was very low (264 U/l). For optimization, we examined enzyme activity in 7 Escherichia coli strains with inducer molecules such as lactose or IPTG. E. coli BL21-CodonPlus(DE3)-RIL exhibited the highest enzyme activity with lactose. Finally, DexT activity was remarkably increased by 12-fold under the optimized culture conditions of a cell density to start induction (OD600) of 0.95, a lactose concentration of 7.5 mM, and an induction temperature of 17oC. These results may effectively apply to the heterologous expression of other large DexT genes.
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KEYWORD
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Dextransucrase, Optimization, Lactose, Response surface methodology, Leuconostoc citreum KM20
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