Construction of methanol sensing Escherichia coli by the introduction of novel chimeric MxcQZ/OmpR two-component system from Methylobacterium organophilum XX

Vidhya Selvamani; Murali Kannan Maruthamuthu; Kulandaisamy Arulsamy; Gyeong Tae Eom; Soon Ho Hong

Korean Journal of Chemical Engineering, Vol.34, No.6, 1734-1739, June, 2017

DOI10.1007/s11814-017-0063-8 Export Citation

Construction of methanol sensing Escherichia coli by the introduction of novel chimeric MxcQZ/OmpR two-component system from Methylobacterium organophilum XX

Vidhya Selvamani¹, Murali Kannan Maruthamuthu¹, Kulandaisamy Arulsamy², Gyeong Tae Eom^{3, 4}, and Soon Ho Hong^{1, †}

¹Department of Chemical Engineering, University of Ulsan, 93 Daehak-ro, Namgu, Ulsan 44610, Korea
²Department of Biotechnology, Indian Institute of Technology Madras, Chennai 600036, India
³Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 45 Jongga-ro, Jung-gu, Ulsan 44429, Korea
⁴Department of Green Chemistry and Environmental Biotechnology, Korea University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon 34144, Korea

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Methylobacterium organophilum XX is a type II facultative methylotroph that can grow on methanol. In M. organophilum XX, the MxcQ/MxcE two-component system (TCS) is involved in methanol metabolism. EnvZ/OmpR in E. coli TCS was exploited to develop a methanol biosensor by engaging the MxcQ/MxcE TCS system. The MxcQZ/ OmpR methanol sensing chimeric TCS was constructed by integrating the sensing domain of M. organophilum MxcQ with the transmitter domain of E. coli EnvZ. The response regulator of the chimeric TCS system is OmpR, which regulates the expression of the ompC and gfp. The expression of ompC was monitored by real-time quantitative PCR analysis. The expression of gfp also confirmed the expression of the ompC. The maximum expression of ompC and gfp occurred with 0.05% of methanol, and the expression started to decline with further increases in methanol concentration. This system delivers rapid detection of methanol in the environment.

Keywords:Biosensors;Chimeric Two-component System;Fluorescence Sensors;Methanol;Synthetic Biology

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[2] Okada T, Nakamura N, Yuasa M, Sekine I, J. Electrochem. Soc., 144(8), 2744 (1997)

[3] Laub MT, Goulian M, Annu. Rev. Genet., 41, 121 (2007)

[4] Stock AM, Robinson VL, Goudreau PN, Annu. Rev. Biochem., 69, 183 (2000)

[5] Mitrophanov AY, Groisman EA, Gen. Dev., 22, 2601 (2008)

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[7] Ganesh I, Ravikumar S, Yoo IK, Hong SH, Bioprocess. Biosyst. Eng., 38, 797 (2014)

[8] Utsumi R, Brissette R, Rampersaud A, Forst S, Oosawa K, Inouye M, Science, 245, 1246 (1989)

[9] Siryaporn A, Goulian M, Mol. Microbiol., 70, 494 (2008)

[10] May T, Okabe, J. Bacteriol., 190, 7479 (2008)

[11] Rogov VV, Rogova NY, Bernhard F, Koglin A, Lohr F, Dotsch V, J. Mol. Biol., 364, 68 (2006)

[12] Rizk SS, Cuneo MJ, Hellinga HW, Science, 15, 1745 (2006)

[13] Chistoserdova L, Chen SW, Lapidus A, Lidstrom ME, J. Bacteriol., 185, 2980 (2003)

[14] Xu HH, Janka JJ, Viebahn M, Hanson RS, Microbiology, 141, 2543 (1995)

[15] Sambrook J, Russell DW, Molecular cloning - A laboratory manual, New York, Cold Spring Harbor (2001).

[16] Basle A, Rummel G, Storici P, Rosenbusch JP, Schirmer T, J. Mol. Biol., 362, 933 (2006)

[17] Bhattacharya D, Cheng J, Proteins, 81, 119 (2013)

[18] Eswar N, Webb B, Marti-Renom MA, Madhusudhan MS, Eramian D, Shen M, Pieper U, Sali A, Current Protocols in Protein Science, Wiley (2007).

[19] Blattner FR, Plunkett G, Bloch CA, Perna NT, Burland V, Riley M, Colladovides J, Glasner JD, Rode CK, Mayhew GF, Gregor J, Davis NW, Kirkpatrick HA, Goeden MA, Rose DJ, Mau B, Shao Y, Science, 277(5331), 1453 (1997)

[20] Forst SA, Roberts DL, Res. Microbiol., 145, 363 (1994)

[21] Eleaume H, Jabbouri S, J. Microbiol. Methods, 59, 363 (2004)

[22] Maruthamuthu MK, Ganesh I, Ravikumar S, Hong SH, Biotechnol. Lett., 37(3), 659 (2015)

[23] Aiba H, Mizuno T, Febs Lett., 261, 19 (1990)

[24] Laemmli UK, Nature, 227, 680 (1970)

[25] Bourque D, Ouellette B, Andre G, Groleau D, Appl. Microbiol. Biotechnol., 37, 7 (1992)

[26] Okoye EL, Ulba BO, Uhobo PC, Oli AN, Ikegbunam MN, J. Scientific Research and Reports, 3, 255 (2013)