E40, a novel microbial protease efficiently detoxifying gluten proteins, for the dietary management of gluten intolerance

• Main Authors: Linda Cavaletti, Anna Taravella, Lucia Carrano, Giacomo Carenzi, Alessandro Sigurtà, Nicola Solinas, Salvatore De Caro, Luigia Di Stasio, Stefania Picascia, Mariavittoria Laezza, Riccardo Troncone, Carmen Gianfrani, Gianfranco Mamone
• Format: Article
• Language: English
• Published: Nature Publishing Group 2019-09-01
• Series: Scientific Reports
• Online Access: https://doi.org/10.1038/s41598-019-48299-7

Abstract Gluten proteins are the causative agent of Celiac Disease (CD), a life-long food intolerance characterized by an autoimmune enteropathy. Inadvertent gluten exposure is frequent even in celiac patients complying with a gluten-free diet, and the supplementation of exogenous gluten-digestive enzymes (glutenases) is indeed a promising approach to reduce the risk of dietary gluten boost. Here we describe Endopeptidase 40, a novel glutenase discovered as secreted protein from the soil actinomycete Actinoallomurus A8, and its recombinant active form produced by Streptomyces lividans TK24. E40 is resistant to pepsin and trypsin, and active in the acidic pH range 3 to 6. E40 efficiently degrades the most immunogenic 33-mer as well as the whole gliadin proteins, as demonstrated by SDS-PAGE, HPLC, LC-MS/MS, and ELISA. T lymphocytes from duodenal biopsies of celiac patients showed a strongly reduced or absent release of IFN-γ when exposed to gluten digested with E40. Data in gastrointestinal simulated conditions suggest that no toxic peptides are freed during gluten digestion by E40 into the stomach to enter the small intestine, thus counteracting the intestinal inflammatory cascade to occur in CD patients. E40 is proposed as a novel candidate in Oral Enzymatic Therapy for the dietary management of gluten toxicity.