Samaneh Khodi, Ali Mohammad Latifi
Baqiyatallah University of Medical Sciences, Tehran, Iran
Posters & Accepted Abstracts: Trends in Green chem
Backgrounds: Pesticides used in controlling harmful populations of insects, can adversely affect human health and cause environmental pollution. The most popular pesticides are organophosphates (OPs) such as Diazinon, Chlorpyrifos, Malathion, etc. Several enzymes have been isolated able to degrade different kinds of OPs such as OPAA (from cytoplasm of Alteromonas sp.) and DFPase (from the brain of squid Loligo vulgaris). To reduce the OP uptake limitation and degrade a broader substrate spectrum with more activity, we demonstrate for the first time a functional codisplay of two distinct hydrolases, DFPase and OPAA, on a single bacterial cell, by fusing each enzyme to N-terminal of InaV (InaV-N). Materials and Methods: To cotranslocate, target genes should be cloned in two different plasmids with two different antibiotic resistances and independent expression strategy, therefore, inaV-N/dfpase and inaV-N/opaa fragments were cloned into pET- 28a(+) and pCDFDuet vectors, separately. Competent E. coli Bl21 cells were transformed by individual recombinant plasmids, as controls, and simultaneously both one by electroporation method. Results: Studies on activity by FPLC with DFPase, OPAA and codisplay cells showed a specific activity of 19.676, 13.125 and 22.46 U/ml, respectively, for Diazinon, and 10.779, 5.213 and 13.784 U/ml, respectively, for Chlorypyrifos. Study on activity by monitoring fluoride-release from DFP by each miligram of DFPase, OPAA and codisplay wet cells showed 500, 250 and 800 Unit, respectively. Discussion: Findings showed a recombinant strain possesses a broader substrate spectrum and a more activity than strains expressing either one of the hydrolases, a whole-cell biocatalyst candidate to detoxify environment polluted by different OPs.
Email: Khodi.sama@gmail.com
Spanish 
Chinese 
Russian 
German 
French 
Japanese 
Portuguese 
Hindi 