Articles & Issues
- Language
- English
- Conflict of Interest
- In relation to this article, we declare that there is no conflict of interest.
- Publication history
-
Received December 22, 2007
Accepted February 27, 2008
-
This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits
unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Copyright © KIChE. All rights reserved.
All issues
Efficient cell surface display of organophosphorous hydrolase using N-terminal domain of ice nucleation protein in Escherichia coli
Department of Chemical Engineering, Pohang University of Science and Technology, Pohang 790-784, Korea 1Key Laboratory of Agricultural Microbiology, School of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, China 2Department of Environmental Science, Kangwon National University, Chuncheon 200-701, Korea 3Department of Biological and Environmental Engineering, Semyung University, Jecheon 390-711, Korea
hjcha@postech.ac.kr
Korean Journal of Chemical Engineering, July 2008, 25(4), 804-807(4), 10.1007/s11814-008-0132-0
Download PDF
Abstract
Recombinant Escherichia coli systems expressing organophosphorous hydrolase (OPH) have been used for detoxifying toxic organophosphate compounds. However, a whole cell biocatalyst system has an intrinsic problem due to substrate diffusion limitation by its cell membrane. As a strategy for reducing this diffusion barrier limitation to enhance whole cell biocatalytic activity, we engineered E. coli cells to target OPH on cell surface using ice nucleation protein (InaK) as a surface targeting motif, especially N-terminal domain of InaK (InaK-N). The whole cell OPH activities of the cells expressing InaK/OPH fusion constructs were higher (~2.5-fold for InaK-N and ~5.7-fold for combined N- and C-terminal domain of InaK (InaK-NC)) than that of the cells expressing cytosolic OPH. Interestingly, the membrane targeting efficiency of the cells expressing InaK-N/OPH fusion proteins was ~2.2-fold higher compared to the cells expressing InaK-NC/OPH even though both whole cell and total cell lysate OPH activities were lower. Therefore, we found that the small size N-terminal domain of InaK is more efficient for targeting OPH on the cell surface, and the surface display of OPH using N-terminal InaK domain can reduce the mass-transfer problem in whole cell bioconversion system.
Keywords
References
Donarski WJ, Dumas DP, Heitmeyer DP, Lewis VE, Raushel FM, Biochemistry, 28, 4650 (1989)
McDaniel CS, Harper LL, Wild JR, J. Bacteriol., 170, 2306 (1988)
Mulbry WW, Karns JS, J. Bacteriol., 171, 6740 (1989)
Grimsley JK, Scholtz JM, Pace CN, Wild JR, Biochemistry, 36, 14366 (1997)
Lai K, Stolowich NJ, Wild JR, Arch. Biochem. Biophys., 318, 59 (1995)
Lewis VE, Donarski WJ, Wild JR, Raushel FM, Biochemistry, 27, 1591 (1988)
Serdar CM, Gibson DT, Biomed. Tech., 3, 567 (1985)
Chen W, Mulchandani A, Trends Biotechnol., 16, 71 (1998)
Mansee AH, Chen W, Mulchandani A, Biotechnol. Bioprocess Eng., 5, 436 (2000)
Kang DG, Lim GB, Cha HJ, J. Biotechnol., 118, 379 (2005)
Kang DG, Choi SS, Cha HJ, Biotechnol. Prog., 22, 406 (2006)
Lee JS, Shin KS, Pan JG, Kim CJ, Nat. Biotechnol., 18, 645 (2000)
Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss R, Nat. Biotechnol., 15, 29 (1997)
Lee JC, Park SY, Choi CY, Chung J, Lee MS, Biotechnol. Bioprocess Eng., 12, 282 (2007)
Richins RD, Kaneva I, Mulchandani A, Chen W, Nat. Biotechnol., 15, 984 (1997)
Sousa C, Cebolla A, Lorenzo V, Nat. Biotechnol., 14, 1017 (1996)
Shimazu M, Mulchandani A, Chen W, Biotechnol. Prog., 17, 76 (2001)
Yoo HJ, Seo JH, Kang DG, Cha HJ, Korean J. Chem. Eng., 24(1), 99 (2007)
Li L, Kang DG, Cha HJ, Biotechnol. Bioeng., 85(2), 214 (2004)
Kang DG, Kim JYH, Cha HJ, Biotechnol. Lett., 24, 879 (2002)
Caldwell SR, Newcomb JR, Schlecht KA, Raushel FM, Biochemistry, 30, 7438 (1991)
McDaniel CS, Harper LL, Wild JR, J. Bacteriol., 170, 2306 (1988)
Mulbry WW, Karns JS, J. Bacteriol., 171, 6740 (1989)
Grimsley JK, Scholtz JM, Pace CN, Wild JR, Biochemistry, 36, 14366 (1997)
Lai K, Stolowich NJ, Wild JR, Arch. Biochem. Biophys., 318, 59 (1995)
Lewis VE, Donarski WJ, Wild JR, Raushel FM, Biochemistry, 27, 1591 (1988)
Serdar CM, Gibson DT, Biomed. Tech., 3, 567 (1985)
Chen W, Mulchandani A, Trends Biotechnol., 16, 71 (1998)
Mansee AH, Chen W, Mulchandani A, Biotechnol. Bioprocess Eng., 5, 436 (2000)
Kang DG, Lim GB, Cha HJ, J. Biotechnol., 118, 379 (2005)
Kang DG, Choi SS, Cha HJ, Biotechnol. Prog., 22, 406 (2006)
Lee JS, Shin KS, Pan JG, Kim CJ, Nat. Biotechnol., 18, 645 (2000)
Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss R, Nat. Biotechnol., 15, 29 (1997)
Lee JC, Park SY, Choi CY, Chung J, Lee MS, Biotechnol. Bioprocess Eng., 12, 282 (2007)
Richins RD, Kaneva I, Mulchandani A, Chen W, Nat. Biotechnol., 15, 984 (1997)
Sousa C, Cebolla A, Lorenzo V, Nat. Biotechnol., 14, 1017 (1996)
Shimazu M, Mulchandani A, Chen W, Biotechnol. Prog., 17, 76 (2001)
Yoo HJ, Seo JH, Kang DG, Cha HJ, Korean J. Chem. Eng., 24(1), 99 (2007)
Li L, Kang DG, Cha HJ, Biotechnol. Bioeng., 85(2), 214 (2004)
Kang DG, Kim JYH, Cha HJ, Biotechnol. Lett., 24, 879 (2002)
Caldwell SR, Newcomb JR, Schlecht KA, Raushel FM, Biochemistry, 30, 7438 (1991)
