Antimicrobial resistance (AMR) poses a critical global health challenge, jeopardizing the efficacy of infectious disease treatments and necessitating innovative solutions. The exploration of plants, renowned for their wealth of bioactive compounds, as sustainable sources of antimicrobials is gaining momentum. Sansevieria, a succulent plant genus, boasts ornamental, medicinal, and pollution-remediating attributes owing to its phytochemical richness. Notably, specific species exhibit inherent antimicrobial properties, positioning them as promising reservoirs of alternative antimicrobial agents. Efficient cultivation methods are imperative for the economically viable production of Sansevierias with potential medicinal benefits. In addressing this need, in vitro micropropagation emerges as a strategic solution, enabling mass multiplication and germplasm conservation. Leveraging our previous success in achieving efficient in vitro regeneration of Sansevieria trifasciata through elevated temperatures and auxin supplementation, we present a novel PTC-assisted antimicrobial elicitation platform. This innovative approach resulted in significantly augmented biomass production, heightened concentrations of potent phytochemicals, and increased antibacterial activity in tissue extracts compared to their field-grown counterparts. Gas chromatography-mass spectrometry (GCMS) profiling confirmed the upregulation of phytochemicals known for their antibacterial efficacy. Notably, Citronellol, 7,8-Epoxylanostan-11-ol, 3-acetoxy, and several new compounds were markedly elevated in PTC-raised Sansevieria trifasciata. Our study furnishes compelling evidence for the efficacy of in vitro techniques in enhancing the growth and phytochemical profiles of Sansevieria plants. This opens new avenues for their commercial production, particularly in addressing the challenges posed by antimicrobial resistance. Importantly, this study represents the first exploration of phytochemical bioprospecting in the Sansevieria genus through in vitro tissue culture endeavors.