Abstract

The use of the organophosphorus hydrolase (OPH) enzyme to degrade Organophosphate compounds is one of the most frequently used decontamination methods. OPH is an similar to 36-kDa homodimericmetalloprotein that is found in the membrane of Flavobacterium sp. strain ATCC 27551 and Brevundimonasdiminuta MG and is capable of hydrolyzing a wide range of oxon and thion such as paraoxon and parathion. The OPH gene (opd) has been expressed in many hosts such as bacteria, insects, insect cells, fungi and Streptomyces spp. High level and soluble expression and correct folding of each protein are important factors. Fusion proteins including TRX, Gb1 and MBP are commonly used to increase solubility, improved folding and in some cases, stability. The present study evaluated OPH expression level, solubility proper folding and stability by cloning the opd gene into pET32a and pET21a and expressing the resulting vectors in E. coli shuffle t7. The pET32a vector encodes a fusion protein containing TRX that is not present in the pET21a. The results reveal increased expression level, moderate solubility, proper folding and stability in produced OPH by the pET32a-opd construct compared to the pET21a vector due to the presence of the fusion with TRX in pET32a vector.