Genomic Characterization and Functional Validation of Microencapsulated Fibrinolytic Proteases Produced By Bacillus pacificus HSFI-5

Abstract Microbial fibrinolytic proteases offer significant potential for breaking down degradation in biotechnology applications. In this study, we used whole-genome sequencing and bioactivity assays to examine microencapsulated extracellular fibrinolytic proteases from Bacillus pacificus HSFI-5. Sequencing on the Illumina and MGI platforms identified at least 7 protease genes, including vpr , aprE , and inha , linked to fibrinolytic activity. Both platforms produced similar genome sizes (about 5.45–5.48 Mb) and GC contents (about 35.3%), showing consistent results. Comparative genomics placed the strain within the B. pacificus group, with matching genome size and GC content across both platforms. We microencapsulated semi-purified extracellular proteases with maltodextrin and Arabic gum, then tested them in BALB/c mice as a model of carrageenan-induced thrombosis. Microencapsulation was intended to improve stability and enabled oral delivery of extracellular proteases while preserving their functional activity in a carrageenan-induced thrombosis model, which served as a biological validation system. Statistical analysis indicated selective modulation of intrinsic coagulation parameters without broad anticoagulant effects. These findings demonstrate how genome-guided enzyme discovery, combined with formulation engineering, can support the functional characterization of microbial proteases with potential applications in biotechnology and in enzyme stabilization strategies.