Efficient cell surface display of organophosphorous hydrolase using N-terminal domain of ice nucleation protein in <Emphasis Type="Italic">Escherichia coli</Emphasis> |
| |
Authors: | Dong Gyun Kang Lin Li Jeong Hyub Ha Suk Soon Choi Hyung Joon Cha |
| |
Affiliation: | (1) Department of Chemical Engineering, Pohang University of Science and Technology, Pohang, 790-784, Korea;(2) Key Laboratory of Agricultural Microbiology, School of Life Science and Technology, Huazhong Agricultural University, Wuhan, 430070, China;(3) Department of Environmental Science, Kangwon National University, Chuncheon, 200-701, Korea;(4) Department of Biological and Environmental Engineering, Semyung University, Jecheon, 390-711, Korea |
| |
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.
This work was presented at 13
th
YABEC symposium held at Seoul, Korea, October 20–22, 2007 |
| |
Keywords: | Cell Surface Display Ice Nucleation Protein N-Terminal Domain Organophosphorus Hydrolase Escherichia coli Whole Cell Biocatalyst |
本文献已被 SpringerLink 等数据库收录! |
|