Advancing a facile one-pot synthetic approach for the fabrication of a hybrid heterojunction photocatalyst remains a significant challenge in research pursuits. Herein, a microspherelike trinary hybrid nanocomposite has been synthesized (NH/PIn/ MAA/Ag). It comprises exfoliated single-and a few-layered Ni(OH) 2 (NH nanosheets), mercaptoacetate-functionalized polyindole (PIn/MAA), and Ag nanoparticles (AgNPs) through an in situ approach. The formation mechanism is based on the exfoliation of stacked Ni(OH) 2 multilayers [i.e., Ni(OH) 2 microflowers] and stabilization of NH nanosheets through host−guest formation of PIn/MAA, followed by the adsorption−reduction of Ag + ions in a one-pot reaction at low temperature. Surface morphological analyses of hybrid nanocomposite microspheres have exhibited that highly dense Ni(OH) 2 microflowers have been transformed into low-density layered forms (NH nanosheets) within the polymeric platform (PIn/MAA) with deposited AgNPs. An interfacial heterojunction has been developed between the components in the depletion region, leading to an improvement in photocatalytic efficiency through a synergistic effect over the components for charge separation and transfer through the heterojunction interface via solid-state mediator Ag-based Z-scheme charge transfer dynamics. The superior photocatalytic degradation of tetracycline (98.2%) by trinary hybrid microspheres can be attributed to the deteriorated recombination rate of electron−hole pairs with reduced charge transfer resistance of the heterojunction in the photocatalyst, as obvious from photoluminescence, electrochemical impedance spectroscopy, chronoamperometry, and timeresolved photoluminescence (TRPL) analyses. Moreover, the antibacterial properties of microspheres against Bacillus pumilus (Gram-positive) and Escherichia coli (Gram-negative) bacteria have validated their potential as promising materials for the overall purification of aquatic systems.