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KMID : 0545120060160091422
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Journal of Microbiology and Biotechnology
2006 Volume.16 No. 9 p.1422 ~ p.1428
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An Efficient Secretion of Type I Secretion Pathway-Dependent Lipase, TliA,in Escherichia coli: Effect of Relative Expression Levels and Timing of Passenger Protein and ABC Transporter
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Eom Gyeong-Tae
Rhee Joon-Shick
Song Jae-Kwang
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Abstract
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An ABC transporter apparatus of the Gram-negative bacterial type I secretion pathway can be used as a secretory protein expression system in Escherichia coli. Four types of coexpression systems for the Pseudomonas fluorescens lipase gene, tliA, and its cognate ABC transporter gene cluster, tliDEF, were constructed. When the relative expression levels were changed by adding different concentrations of IPTG, the secretion (16.9 U/ml of culture) of TliA in E. coli [pTliDEFA-223+pACYC184] was significantly higher than E. coli [pKK223-3+pTliDEFA-184] secreting the lowest level of TliA (5.2 U/ml of culture). Maximal accumulation of the lipase secreted occurred in the mid-exponential phase, implying that the efficient protein secretion via an ABC transporter was restricted only to actively growing cells. Finally, the secretion level of TliA in E. coli [pTliDEFA-223+pACYC184] was increased to 26.4 U/ml by inducing gene expression at the culture initiation time. These results indicate that a significant increase in the ABC transporter-dependent protein secretion can be achieved by simply controlling the relative expression levels between the ABC transporter and its passenger protein, even in the recombinant E. coli cells.
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KEYWORD
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ABC transporter, Escherichia coli, lipase, type I secretion pathway
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