Abstract
HIV-1 is an infectious virus that causes acquired immunodeficiency syndrome (AIDS) and it isone of the major causes of deaths worldwide. The production of HIV-1 protease (PR) on alarge scale has been a problem forscientists due to its cytotoxicity, low yield, insolubility, and low activity. HIV-1C-SA protease has been cloned, expressed, and purified previously, however, with low recovery (0.25mg/L). Herein we report an optimal expression and solubilisation procedure to recover active HIV-1C-SA protease enzyme from inclusion bodies. The HIV protease was expressed in seven different vectors (pET11b, pET15b, pET28a pET32a, pET39b, pET41b and pGEX 6P-1). The highest expression was achieved when the vectorpET32a (Trx tag) was employed. A total of 19.5mg of fusion protein was refolded of which 5.5mg of active protease was obtained after cleavage. The free protease had a high specific activity of 2.81 µmoles/min/mg. Interestingly the Trx-fusion protein also showed activity closer (1.24 µmoles/min/mg) to that of the free protease suggesting that thepET32a vector (Trx tag) expressed in BL21(DE3) pLysS provides a more efficientway to obtain HIV-1 protease.
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