Siropins, novel serine protease inhibitors from gut microbiota acting on human proteases involved in inflammatory bowel diseases

Autor: Samira Boudebbouze, Florette Szukala, Vincent Mariaule, Ali Gargouri, Emmanuelle Maguin, Nizar Akermi, Nicolas Pons, Héla Mkaouar, Josan Marquez, Nadia Gaci, Moez Rhimi
Přispěvatelé: MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Centre de Biotechnologie de Sfax (CBS), Grenoble Outstation, European Molecular Biology Laboratory, MetaGenoPolis, Institut National de la Recherche Agronomique (INRA), Département Microbiologie et Chaîne Alimentaire (MICA), Agence Nationale de la Recherche (ANR) [14-CE16-0018], project CMCU-PHC Utique-Campus France [14G0816, 30666QM], Microbiota Interaction with Human and Animal (MIHA), Institut National de la Recherche Agronomique (INRA)-AgroParisTech-Institut National de la Recherche Agronomique (INRA)-AgroParisTech, AgroParisTech-Université Paris-Saclay-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)
Jazyk: angličtina
Rok vydání: 2016
Předmět:
Zdroj: Microbial Cell Factories
Microbial Cell Factories, BioMed Central, 2016, 15, pp.1-13. ⟨10.1186/s12934-016-0596-2⟩
Microbial Cell Factories, BioMed Central, 2016, 15 (1), ⟨10.1186/s12934-016-0596-2⟩
Microbial Cell Factories (15), 1-13. (2016)
ISSN: 1475-2859
DOI: 10.1186/s12934-016-0596-2⟩
Popis: Background In eukaryotes, the serpins constitute a wide family of protease inhibitors regulating many physiological pathways. Many reports stressed the key role of serpins in several human physiopathologies including mainly the inflammatory bowel diseases. In this context, eukaryotic serpins were largely studied and their use to limit inflammation was reported. In comparison to that, bacterial serpins and mainly those from human gut microbiota remain poorly studied. Results The two genes encoding for putative serpins from the human gut bacterium Eubacterium sireaum, display low sequence identities. These genes were overexpressed and the encoded proteins, named Siropins, were purified. Activity studies demonstrated that both purified proteins inhibited serine proteases but surprisingly they preferentially inhibited two human serine proteases (Human Neutrophil Elastase and Proteinase3). The biochemical characterization of these Siropins revealed that Siropin 1 was the most active and stable at low pH values while Siropin 2 was more thermoactive and thermostable. Kinetic analysis allowed the determination of the stoichiometry of inhibition (SI) which was around 1 and of the association rate constants of 7.7 × 104 for the Human Neutrophil Elastase and 2.6 × 105 for the Proteinase3. Moreover, both Siropins displayed the ability to inhibit proteases usually present in fecal waters. Altogether our data indicate the high efficiency of Siropins and their probable involvement in the control of the overall intestine protease activity. Conclusions Here we report the purification and the biochemical characterization of two novel serpins originated from Eubacterium sireaum, a human gastro-intestinal tract commensal bacteria. These proteins that we called Siropins, efficiently inhibited two human proteases reported to be associated with inflammatory bowel diseases. The determination of the biochemical properties of these enzymes revealed different temperature and pH behaviours that may reflect adaptation of this human commensal bacterium to different ecological environments. To the best of our knowledge, it is the first bacterial serpins showing an attractive inhibition of fecal proteases recovered from a mice group with chemically induced inflammation. Altogether our data highlight the interesting potential of Siropins, and serpins from the human gut microbiota in general, to be used as new alternative to face inflammatory diseases. Electronic supplementary material The online version of this article (doi:10.1186/s12934-016-0596-2) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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