@article{open3465,
          volume = {42},
          number = {3},
           month = {February},
          author = {Devtulya Chander and Diksha Koul and Shubham Singh and Ravi S. Manhas and Diljeet Kumar and Sanket Shukla and Ravi P. N. Mishra and Zabeer Ahmed and Asha Chaubey},
            note = {Copyright of this data belongs to WORLD JOURNAL OF MICROBIOLOGY \& BIOTECHNOLOGY},
         address = {VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS},
           title = {Heterologous expression, purification and functional characterization of recombinant serratiopeptidase from Serratia marcescens AD-W2},
       publisher = {SPRINGER},
            year = {2026},
         journal = {WORLD JOURNAL OF MICROBIOLOGY \& BIOTECHNOLOGY},
        keywords = {{\ensuremath{<}}italic{\ensuremath{>}}E.coli{\ensuremath{<}}/italic{\ensuremath{>}} K12; serratiopeptidase; Protein refolding; Tangential flow filtration; Anti-inflammatory activity},
             url = {http://crdd.osdd.net/open/3465/},
        abstract = {Serratiopeptidase, a proteolytic enzyme with therapeutic applications, is traditionally produced from the bacterium Serratia marcescens. Recombinant production of serratiopeptidase in Escherichia coli offers a safer alternative to the biosafety concerns of the producer. Present study involves cloning and heterologous expression of thermoactive serratiopeptidase gene from S. marcescens AD-W2 in E. coli K12 in pET28a plasmid. Optimized expression conditions i.e. 37 degrees C, OD600 5, 5\% L-rhamnose, 5mM IPTG, and 50\% dissolved oxygen led to the final yield of 190 mg/g serratiopeptidase (4747 mg protein/L within 9 h in a bioreactor. Purification and refolding of recombinant serratiopeptidase was performed in a single step using Tangential Flow Filtration (TFF) and diafiltration process. The purified recombinant serratiopeptidase exhibited specific activity of 1800 Units/mg protein, with an optimal activity at pH 9.0 and temperature 50 degrees C. The value of kinetic constant Km was calculated as 1.38 mg/mL for casein. The recombinant serratiopeptidase demonstrated comparable anti-inflammatory activity to the commercially available serratiopeptidase, inhibiting nitric oxide release and pro-inflammatory cytokine production in LPS-stimulated murine macrophage cell line RAW 264.7. The study reveals that the recombinant serratiopeptidase produced in E. coli K12 holds promising source of safe and effective anti-inflammatory agent.}
}