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Food preservation using biopreservative agents mainly microbial based metabolites have gained more attention in the modern food sector due to their broad spectrum anti-microbial efficacy with high biocompatibility. However, the effectiveness of these biopreservatives is highly dependent on suitable formulation which will enhance the desired efficacy without affecting the nutritional value of the foods. Polylysine-actinomycete derived anti-microbial metabolite is used as biopreservative agent in various food products. This review provides a valuable insight into the formulation of polylysine with nanotechnology principles for enhanced antimicrobial efficacy against food spoilage organisms. This review highlighted the diverse methods of polylysine nanoformulation preparation. Formulation of polylysine as functionalised nanomaterials can be prepared via green science or green technology principles was extensively studied in this review. This study also revealed the methods for the determination of anti-microbial assays and biocompatibility tests in vitro and in vivo biocompatibility tests. Future prospects of nanoformulated polylysine in bio preservation can be extended to safe processing of canned and ready-to-eat food products without affecting the nutritional value of the foods.