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Antibiotic resistance is a growing concern today that affects the entire world, necessitating the
development of new antibacterial medications. Nosocomial infections pose a serious challenge for
patients and hinder effective treatment. The second most common cause of nosocomial infections
is Enterococcus faecalis, and it depends on the lactate dehydrogenase enzyme's capacity to
maintain redox balance for growth, resistance, and virulence. As demonstrated before by
computational technique, our study attempted to assess the antibacterial effect of Gossypol on
Enterococcus faecalis by inhibiting Lactate Dehydrogenase Enzyme. We used six different
stressors, including 0.01% SDS, 2.5mM H2O2, 8% Ethanol, 10% DMSO, 10% Glucose, and
0.25% HOCl, in addition to our inhibitor Gossypol. Gossypol was employed in the following
concentrations: 5, 10, 25, 50, 75, 100, 150, 200 micrograms per milliliter using a large test tube
method, absorbance on a UV-visible spectrophotometer, and on a microscale using a 96-well flatbottom plate with a Microplate reader. Between control (Culture Media) and vehicle control, which
is DMSO, there is no discernible difference. At higher dosages of Gossypol, such as 100 and 200
micrograms per milliliter, there is significant growth inhibition; nevertheless, 5, 10, 25, 50, 75, and
100 micrograms per milliliter show no significant inhibition. Minimum Inhibitory Concentration
is 100 microgram per milliliter. We used our six different stress factors with MIC value of
Gossypol. There is no discernible growth inhibition when Gossypol is employed in conjunction
with stress factors such 0.01% SDS, 2.5mM H2O2, and 0.25% HOCl. However, at 100
micrograms per milliliter at the fourth and fifth hours, glucose 10% exhibits a strong inhibitory
impact, though not by a great deal. Along with MIC value of Gossypol, 100 micrograms per
milliliter, 8% ethanol and 10% DMSO significantly slowed the development of the bacteria. The
lactate dehydrogenase enzyme in Enterococcus faecalis was effectively inhibited by gossypol.
Future in vivo studies are required to demonstrate the antibacterial activity of Gossypol in greater
detail, as well as to compare it to antibiotics as an adjunctive treatment.