Efficient cell surface display of organophosphorous hydrolase using N-terminal domain of ice nucleation protein in Escherichia coli
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- Title
- Efficient cell surface display of organophosphorous hydrolase using N-terminal domain of ice nucleation protein in Escherichia coli
- Authors
- Kang, DG; Li, L; Ha, JH; Choi, SS; Cha, HJ
- Date Issued
- 2008-07
- Publisher
- KOREAN INST CHEM ENGINEERS
- 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 (similar to 2.5-fold for InaK-N and similar to 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 similar to 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
- cell surface display; ice nucleation protein; N-terminal domain; organophosphorus hydrolase; Escherichia coli; whole cell biocatalyst; PSEUDOMONAS-DIMINUTA; PERIPLASMIC SECRETION; PHOSPHOTRIESTERASE; HYDROLYSIS; GENE; BIODEGRADATION; DETOXIFICATION; PESTICIDES; PATHWAY; CLONING
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/29410
- DOI
- 10.1007/s11814-008-0132-0
- ISSN
- 0256-1115
- Article Type
- Article
- Citation
- KOREAN JOURNAL OF CHEMICAL ENGINEERING, vol. 25, no. 4, page. 804 - 807, 2008-07
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