Enhanced biodegradation of toxic organophosphate compounds using recombinant Escherichia coli with sec pathway-driven periplasmic secretion of organophosphorus hydrolase.
Although Escherichia coli can be genetically engineered to degrade environmental toxic organophosphate compounds (OPs) to nontoxic materials, a critical problem in such whole cell systems is limited substrate diffusion. The present work examined whether periplasmic expression of organophosphorus hydrolase (OPH) resulted in better whole cell enzymatic activity compared to standard cytosolic expression. Recombinant OPH periplasmic expression was achieved using the general secretory (sec) pathway with the pelB signal sequence. We found that while total OPH activity in periplasmic-expressing cell lysates was lower compared to that in cytosolic-expressing cell lysates whole cell OPH activity was 1.8-fold greater at 12 h post-induction in the periplasmic-expressing cells as a result of OPH translocation into the periplasmic space ( approximately 67% of whole cell OPH activity was found in the periplasmic fraction). These data suggest that E. coli engineered to periplasmically secrete OPH via the sec pathway may provide an improved whole cell biodegradation system for destruction of environmental toxic OPs.
Kang
Dong Gyun
DG
Department of Chemical Engineering & Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
Choi
Suk Soon
SS
Cha
Hyung Joon
HJ
D043303:Aryldialkylphosphatase / Q000378:metabolism*; D004926:Escherichia coli / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics; D009154:Mutation / Q000235:genetics; D009943:Organophosphorus Compounds / Q000378:metabolism* / Q000633:toxicity; D019897:Periplasm / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics
D009943:Organophosphorus Compounds; D043303:Aryldialkylphosphatase