Ramlucken, UraishaKrishna, Suresh Babu NaiduGovender, Patrick2022-01-132022-01-132021-12-31Ramlucken, U., Babu Naidu, K.S. and Govender, P. 2021.Improved production of HIV-1 subtype C protease from transgenic E. Coli. The Open Microbiology Journal. 15(1): 168-176. doi:10.2174/18742858021150101681874-2858https://hdl.handle.net/10321/3763Background: Human Immunodeficiency Virus 1 (HIV-1) subtype C is responsible for the majority of infections of patients in Southern Africa. The HIV protease is a primary target for the development of highly efficient anti-retroviral pharmaceuticals because of its pivotal role in the maturation of the virus in the host cell. For target validation of novel HIV protease inhibitors, there is a need for the availability of an abundance of this protease. Objective: This study reports an optimized method to produce HIV-1 protease derived from HIV-1 subtype C. Methods: It involves the use of a transgenic E. colistrain that overexpresses the native form of the enzyme <jats:italic>via</jats:italic> inclusion bodies. A stringent method for the isolation, purification, and renaturation resulted in the production of highly pure active HIV-1 protease. In order to facilitate an increase in protease yields, an optimized growth strategy was developed. In this regard, a chemically defined medium with lower glucose content and devoid of essential amino acids of the TCA cycle was used as an alternative to the widely used nutrient-rich Luria Bertani (LB) medium. Results: Results indicated an increase in protease yield up to twice the amount, thereby making this medium an attractive alternative for increasing biomass and HIV protease production for future research. Conclusion: An optimized method for HIV-1 protease derived from HIV-1 subtype C production using chemically defined media was established. This was achieved using a known method to isolate and purify the enzyme with the use of a specialized feeding strategy.9 p.en0707 Veterinary SciencesE. coliHIV proteaseIon exchange chromatographyOptimizationSulfate-polyacrylamide gelElectrophoresis.Article2022-01-1110.2174/1874285802115010168