2. Academic Validation
  3. A putative cytotoxic serine protease from Salmonella typhimurium UcB5 recovered from undercooked burger

A putative cytotoxic serine protease from Salmonella typhimurium UcB5 recovered from undercooked burger

  • Sci Rep. 2023 Mar 9;13(1):3926. doi: 10.1038/s41598-023-29847-8.
Essam Kotb 1 2,  Baher A El-Nogoumy 3,  Haifa A Alqahtani 4,  Asmaa A Ahmed 5,  Hussah A Al-Shwyeh 4 6,  Sakina M Algarudi 4 6,  Hanan Almahasheer 4
Affiliations

Affiliations

  • 1 Department of Biology, College of Science, Imam Abdulrahman Bin Faisal University (IAU), P.O. Box 1982, Dammam, 31441, Saudi Arabia. ekotb@hotmail.com.
  • 2 Basic & Applied Scientific Research Center, Imam Abdulrahman Bin Faisal University, P.O. Box 1982, Dammam, 31441, Saudi Arabia. ekotb@hotmail.com.
  • 3 Department of Botany and Microbiology, Faculty of Science, Kafrelsheikh University, Kafrelsheikh, 33516, Egypt.
  • 4 Department of Biology, College of Science, Imam Abdulrahman Bin Faisal University (IAU), P.O. Box 1982, Dammam, 31441, Saudi Arabia.
  • 5 Department of Statistics, Faculty of Commerce, Al-Azhar University (Girls' Branch), P.O. Box 11751, Cairo, Egypt.
  • 6 Basic & Applied Scientific Research Center, Imam Abdulrahman Bin Faisal University, P.O. Box 1982, Dammam, 31441, Saudi Arabia.
PMID: 36894576 DOI: 10.1038/s41598-023-29847-8
Abstract

A putative virulence exoprotease designated as UcB5 was successfully purified from the bacterium Salmonella typhimurium to the electrophoretic homogeneity with 13.2-fold and 17.1% recovery by hydrophobic, ion-exchange, and gel permeation chromatography using Phenyl-Sepharose 6FF, DEAE-Sepharose CL-6B, and Sephadex G-75, respectively. By applying SDS-PAGE, the molecular weight was confirmed at 35 kDa. The optimal temperature, pH, and isoelectric point were 35 °C, 8.0, 5.6 ± 0.2, respectively. UcB5 was found to have a broad substrate specificity against almost all the tested chromogenic substrates with maximal affinity against N-Succ-Ala-Ala-Pro-Phe-pNA achieving Km of 0.16 mM, Kcat/Km of 3.01 × 105 S-1 M-1, and amidolytic activity of 28.9 µmol min-1 L-1. It was drastically inhibited by TLCK, PMSF, SBTI, and aprotinin while, DTT, β-mercaptoethanol, 2,2'-bipyridine, o-phenanthroline, EDTA, and EGTA had no effect, which suggested a serine protease-type. Also, it has shown a broad substrate specificity against a broad range of natural proteins including serum proteins. A cytotoxicity and electron microscopy study revealed that UcB5 could cause subcellular proteolysis that finally led to liver necrosis. For this, future research should focus on using a combination of external antiproteases and antimicrobial agents for the treatment of microbial diseases instead of using drugs alone.

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