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@ARTICLE{Chander2026-mk,
  title     = "Heterologous expression, purification and functional
               characterization of recombinant serratiopeptidase from Serratia
               marcescens {AD-W2}",
  author    = "Chander, Devtulya and Koul, Diksha and Singh, Shubham and
               Manhas, Ravi S and Kumar, Diljeet and Shukla, Sanket and Mishra,
               Ravi P N and Ahmed, Zabeer and Chaubey, Asha",
  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 °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 °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.",
  journal   = "World J. Microbiol. Biotechnol.",
  publisher = "Springer Science and Business Media LLC",
  volume    =  42,
  number    =  3,
  month     =  feb,
  year      =  2026,
  keywords  = "E.coli K12; serratiopeptidase; Anti-inflammatory activity;
               Protein refolding; Tangential flow filtration",
  copyright = "https://www.springernature.com/gp/researchers/text-and-data-mining",
  language  = "en"
}