Coexpression of molecular chaperone enhances activity and export of organophosphorus hydrolase in Escherichia coli

Title
Coexpression of molecular chaperone enhances activity and export of organophosphorus hydrolase in Escherichia coli
Author(s)
서정현강동균[강동균]김창섭김임규[김임규]최석순[최석순]하정협[하정협]남수완[남수완]임근배[임근배]차형준[차형준]
Keywords
ICE NUCLEATION PROTEIN; ARGININE TRANSLOCATION PATHWAY; CELL-SURFACE DISPLAY; N-TERMINAL DOMAIN; PERIPLASMIC SECRETION; PSEUDOMONAS-DIMINUTA; TAT PATHWAY; PHOSPHOTRIESTERASE; HYDROLYSIS; MEMBRANES
Issue Date
201208
Publisher
WILEY-BLACKWELL
Citation
BIOTECHNOLOGY PROGRESS, v.28, no.4, pp.925 - 930
Abstract
Periplasmic secretion has been used in attempts to construct an efficient whole-cell biocatalyst with greatly reduced diffusion limitations. Previously, we developed recombinant Escherichia coli that express organophosphorus hydrolase (OPH) in the periplasmic space using the twin-arginine translocation (Tat) pathway to degrade environmental toxic organophosphate compounds. This system has the advantage of secreting protein into the periplasm after folding in the cytoplasm. However, when OPH was expressed with a Tat signal sequence in E. coli, we found that the predominant OPH was an insoluble premature form in the cytoplasm, and thus, the whole-cell OPH activity was significantly lower than its cell lysate activity. In this work, we, for the first time, used a molecular chaperone coexpression strategy to enhance whole-cell OPH activity by improving the periplasmic translocation of soluble OPH. We found that the effect of GroEL-GroES (GroEL/ES) assistance on the periplasmic localization of OPH was secretory pathway dependent. We observed a significant increase in the amount of soluble mature OPH when cytoplasmic GroEL/ES was expressed; this increase in the amount of mature OPH might be due to enhanced OPH folding in the cytoplasm. Importantly, the whole-cell OPH activity of the chaperonecoexpressing cells was similar to 5.5-fold greater at 12 h after induction than that of cells that did not express the chaperone as a result of significant Tat-based periplasmic translocation of OPH in the chaperonecoexpressing cells. Collectively, these data suggest that molecular chaperones significantly enhance the whole-cell activity of periplasmic OPH-secreting cells, yielding an effective whole-cell biocatalyst system with highly reduced diffusion limitations. (c) 2012 American Institute of Chemical Engineers Biotechnol. Prog., 28: 925930, 2012
URI
http://hdl.handle.net/YU.REPOSITORY/27556http://dx.doi.org/10.1002/btpr.1556
ISSN
8756-7938
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공과대학 > 화학공학부 > Articles
이과대학 > 화학생화학부 > Articles
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