Heterologous expression, purification and functional characterization of recombinant serratiopeptidase from Serratia marcescens AD-W2

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).

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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.

Item Type:	Article
Additional Information:	Copyright of this data belongs to WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
Uncontrolled Keywords:	<italic>E.coli</italic> K12; serratiopeptidase; Protein refolding; Tangential flow filtration; Anti-inflammatory activity
Subjects:	Q Science > QR Microbiology
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	10 Jul 2026 01:43
Last Modified:	10 Jul 2026 01:43
URI:	http://crdd.osdd.net/open/id/eprint/3465

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