Desmoplasia is a hallmark feature of pancreatic ductal adenocarcinoma (PDAC) that contributes significantly to treatment resistance. Approaches to enhance drug delivery into fibrotic PDAC tumors continues to be an important unmet need. In this study, we have engineered a tumor-colonizing, E.coli-based agent that expresses both collagenase and hyaluronidase as a strategy to reduce desmoplasia and enhance intratumoral perfusion of anticancer agents. Overall, we observed that the tandem expression of both these enzymes by tumor-colonizing E.coli resulted in the reduced presence of intratumoral collagen and hyaluronan, which likely contributed to enhanced chemotherapeutic efficacy observed when used in combination. These results highlight the importance of combination treatments involving the depletion of desmoplastic components in PDAC prior to, or during, treatment.