Discovery and characterization a new collagenase from Hathewaya massiliensis for effective subcutaneous adipolysis.

Collagenases hydrolyze peptide bonds in collagen and have established therapeutic applications in connective tissue disorders and medical aesthetics, including treatment of Peyronie's disease, Dupuytren's contracture and localized adipose accumulation. Here, we describe HmCol, a novel collagenase from Hathewaya massiliensis. A rationally truncated, codon optimized HmCol gene was cloned into pET30a and expressed in Escherichia coli BL21(DE3). The purified enzyme exhibits robust collagenolytic activity, with an optimum at pH 7.5 and 45 °C, underscoring its clinical potential. A single site mutant, HmColM, retains substrate affinity but displays a five-fold reduction in specific activity compared with wild type HmCol. In vitro assays on porcine skin explants show that HmCol induces rapid and dose dependent adipocyte lysis, whereas HmColM produces a milder effect. These results expand the collagenase toolkit, offering HmCol as a highly active enzyme and HmColM as a tunable variant for next generation therapeutic strategies.