TY - JOUR ID - 88354 TI - The Efficient Solubilization and Refolding of Recombinant Organophosphorus Hydrolyses Inclusion Bodies Produced in Escherichia coli JO - Journal of Applied Biotechnology Reports JA - JABR LA - en SN - 2322-1186 AU - Mirhosseini, Seyed Ali AU - Latifi, Ali Mohammad AU - Mahmoodzadeh Hosseini, Hamideh AU - Seidmoradi, Rezvan AU - Aghamollaei, Hossein AU - Farnoosh, Gholamreza AD - Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran AD - Applied Biotechnology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran AD - Faculty of Biological Science, Alzahra University, Tehran, Iran AD - Chemical Injuries Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran Y1 - 2019 PY - 2019 VL - 6 IS - 1 SP - 20 EP - 25 KW - Organophosphorus Hydrolases KW - Inclusion bodies KW - Solubilization KW - Refolding KW - Protein Activity DO - 10.29252/JABR.06.01.04 N2 - Introduction: Organophosphorus hydrolase(OPH) is an enzyme that can degrade organophosphorus compounds in pesticides. High expression levels of OPH in Escherichia coli lead to form inclusion body in cytoplasmic space which is an inactive form of protein and needs a solubilizing and refolding process. The aim of this study was to compare different methods for solubilization and refolding of recombinant OPH expressed in E. coli. Materials and Methods: OPH was expressed in E. coli and purified by the Ni-NTA column. The refolding efficiency of this protein was assessed by 4 strategies: dialysis, rapid dilution, on column and combination of rapid dilution and dialysis. In each case, the refolding efficiency was evaluated by SDS-PAGE analysis and enzyme activity assay and was compared to find the best procedure. Results: The refolding efficiency of these 4 strategies was estimated at about 12%, 10%, 14% and 50% for on column, rapid dilution, dialysis and combination of rapid dilution and dialysis, respectively. Results showed that during the refolding process, most proteins did not reach the correct structure and aggregated again while the combination of 2 methods, rapid dilution and dialysis provided an appropriate procedure to refold. Conclusions: The combination of rapid dilution and dialysis is an efficient method for refolding OPH. The efficacy of this method for refolding other recombinant proteins can be further investigated. UR - https://www.biotechrep.ir/article_88354.html L1 - https://www.biotechrep.ir/article_88354_09f730f3255de25808a1d00cd3114e3a.pdf ER -