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Mycobacterium tuberculosis (Mtb) expresses a broad-spectrum β-lactamase (BlaC) that mediates resistance to one of the highly effective antibacterials, β-lactams. Nonetheless, β-lactams showed mycobactericidal activity in combination with β-lactamase inhibitor, clavulanate (Clav). However, the mechanistic aspects of how Mtb responds to β-lactams such as Amoxicillin in combination with Clav (referred as Augmentin [AG]) are not clear. Here, we identified cytoplasmic redox potential and intracellular redox sensor, WhiB4, as key determinants of mycobacterial resistance against AG. Using computer-based, biochemical, redox-biosensor, and genetic strategies, we uncovered a functional linkage between specific determinants of β-lactam resistance (e.g. β-lactamase) and redox potential in Mtb. We also describe the role of WhiB4 in coordinating the activity of β-lactamase in a redox-dependent manner to tolerate AG. Disruption of WhiB4 enhances AG tolerance, whereas overexpression potentiates AG activity against drug-resistant Mtb. Our findings suggest that AG can be exploited to diminish drug-resistance in Mtb through redox-based interventions. DOI: http://dx.doi.org/10.7554/eLife.25624.001, eLife digest A bacterium called Mycobacterium tuberculosis causes tuberculosis in humans. Multiple antibiotics are available to treat this infection, yet around one million people still die from tuberculosis each year. One of the reasons that the number of deaths is so high is because many M. tuberculosis cells have become resistant to these drugs. Therefore, new drug treatments are urgently needed to tackle the disease. When cells are under stress – for example, when a bacterial cell is exposed to an antibiotic – they can increase the production of chemicals known as reactive oxygen species. These chemicals are vital to many processes in cells, but if their levels get too high they can kill cells by damaging DNA and other molecules. To prevent this damage, bacterial cells produce molecules, such as mycothiol, to neutralize the excess reactive oxygen species. A therapy called Augmentin is used to fight many different types of bacterial infection. It combines an antibiotic known as amoxicillin with another drug that blocks the activity of a bacterial enzyme responsible for breaking down amoxicillin-like drugs. Augmentin can also kill M. tuberculosis cells, but it was not clear exactly how it works, or how the bacteria might be able to develop resistance to this treatment. Here, Mishra et al. combined a computational technique known as network analysis with experiments to study the affect of Augmentin on M. tuberculosis. The experiments reveal that M. tuberculosis cells can develop resistance to Augmentin by increasing the production of an enzyme that breaks down the antibiotic and by neutralizing reactive oxygen species with help of mycothiol. Augmentin treatment can decrease the production of a protein called WhiB4 in the bacteria. This protein is involved in detecting when cells are stressed and regulates the levels of both mycothiol and the enzyme that breaks down the antibiotic. Increasing the production of this protein made the bacterial cells more susceptible to Augmentin treatment by decreasing the levels of active mycothiol and reducing the production of the enzyme that breaks down the antibiotic drug. These findings suggest that Augmentin could be more effective against drug-resistant tuberculosis and other bacterial infections if it is combined with a drug that can alter the levels of reactive oxygen species inside bacterial cells. The next step is to search for new molecules that may be able to perform such a role. DOI: http://dx.doi.org/10.7554/eLife.25624.002