RESEARCH ARTICLE
Improved Production of HIV-1 Subtype C Protease from Transgenic E. Coli
Uraisha Ramlucken1, Krishna Suresh Babu Naidu2, Patrick Govender1, *
Article Information
Identifiers and Pagination:
Year: 2021Volume: 15
Issue: Suppl-1, M3
First Page: 168
Last Page: 176
Publisher ID: TOMICROJ-15-168
DOI: 10.2174/1874285802115010168
Article History:
Received Date: 08/09/2020Revision Received Date: 02/12/2020
Acceptance Date: 10/12/2020
Electronic publication date: 31/12/2021
Collection year: 2021
open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Background:
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. coli strain that overexpresses the native form of the enzyme via 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.
