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Received July 30, 2006
Accepted August 30, 2006
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Correlation analysis for non-invasive quantitative monitoring of biological activity of recombinant enzyme using green fluorescence protein in Escherichia coli under various culture environments
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea
hjcha@postech.ac.kr
Korean Journal of Chemical Engineering, January 2007, 24(1), 99-101(3), 10.1007/s11814-007-5016-1
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Abstract
Monitoring of biological activity for target enzyme is important for its production in recombinant expression systems. Previously, we demonstrated that green fluorescent protein (GFP) as a fusion partner is successfully tooled for facile, in vivo, and non-invasive quantification of target enzyme levels based on a linear relationship between GFP fluorescence and enzyme (chloramphenicol acetyltransferase; CAT) activity. Here, we investigated the effects of culture environmental variations (initial glucose amount, surface aeration, and inducer concentration) on correlation between GFP fluorescence and CAT activity, and established a general linear correlation as a unique criterion for quantitative monitoring of CAT biological activity. This general correlation for GFP fusion strategy can be applied for non-invasive and on-line monitoring of recombinant enzyme production under various culture conditions without further experimental calibrations.
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References
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Cha HJ, Dalal NG, Vakharia VN, Bentley WE, J. Biotechnol., 69, 9 (1999)
Cha HJ, Dalal NG, Pham MQ, Vakharia VN, Rao G, Bentley WE, Biotechnol. Bioeng., 65(3), 316 (1999)
Cha HJ, Wu CF, Valdes JJ, Rao G, Bentley WE, Biotechnol. Bioeng., 67(5), 565 (2000)
Cha HJ, Shin HS, Lim HJ, Cho HS, Dalal NG, Pham MQ, Bentley WE, Biochem. Eng. J., 24, 225 (2005)
Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC, Science, 263(5148), 802 (1994)
Crameri A, Whitehorn EA, Tate E, Stemmer WPC, Nat. Biotechnol., 14, 315 (1996)
Glick BR, Biotechnol. Adv., 13, 247 (1995)
Kain S, Gauguly S, in F. M. Ausubel et al. (Ed.), Current protocols in molecular biology, John Wiley & Sons, New York, 9.6.1-9.6.12 (1995)
Rodriquez RL, Tait RC, Recombinant DNA techniques: An introduction, Benjamin/Cummings Publishing, Menlo Park, 187-191 (1983)
