The present study employed response surface methodology (RSM) to optimise the prevention of late blowing defects in cheese during storage. The aim was to enhance the inhibition of Clostridium sporogenes in cheese by manipulating three independent variables: Lactiplantibacillus plantarum (utilising two different strains, labelled as A and B, the total cheese inoculation rate was 2% overall and the A : B ratio ranging from 25% to 75%), and lysozyme (ranging from 0 to 0.2 mg·L-1). The response variables considered in this model cheese study included the Clostridium count, pH, and titratable acidity. The results showed that the optimal conditions for inhibiting C. sporogenes and preventing late blowing defects in cheese were achieved with an L. plantarum A : B ratio of 49.54 : 50.46%, nisin at a concentration of 1.762 mg·L-1, and lysozyme at 0.2 mg·L-1. These results demonstrated not only effective inhibition of C. sporogenes, a pivotal contributor to late blowing defects in cheese but also indicated favourable outcomes in terms of acidity parameters, which are crucial quality criteria for cheese production. The application of Response Surface Methodology revealed that late blowing defects can be prevented using relatively lower concentrations of antimicrobial agents, along with a judicious selection of appropriate cultures. This research highlights the potential for more efficient and costeffective strategies to maintain cheese quality by minimising the risk of late blowing defects.