creators_name: Chander, Devtulya creators_name: Koul, Diksha creators_name: Singh, Shubham creators_name: Manhas, Ravi S. creators_name: Kumar, Diljeet creators_name: Shukla, Sanket creators_name: Mishra, Ravi P. N. creators_name: Ahmed, Zabeer creators_name: Chaubey, Asha type: article datestamp: 2026-07-10 01:43:41 lastmod: 2026-07-10 01:43:41 metadata_visibility: show title: Heterologous expression, purification and functional characterization of recombinant serratiopeptidase from Serratia marcescens AD-W2 ispublished: pub subjects: QR keywords: E.coli K12; serratiopeptidase; Protein refolding; Tangential flow filtration; Anti-inflammatory activity note: Copyright of this data belongs to WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 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. date: 2026-02-24 date_type: published publication: WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY volume: 42 number: 3 publisher: SPRINGER place_of_pub: VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS refereed: TRUE official_url: https://link.springer.com/article/10.1007/s11274-026-04795-3 citation: 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 (2026) Heterologous expression, purification and functional characterization of recombinant serratiopeptidase from Serratia marcescens AD-W2. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY, 42 (3). document_url: http://crdd.osdd.net/open/3465/1/references.bib