title: Feruloyl esterase activity is influenced by bile, probiotic intestinal adhesion and milk fat.
abstract: Cinnamoyl esterases (CE) are microbial and mammalian intestinal enzymes able to release antioxidant hydroxycinnamic acids from their non-digestible ester-linked forms naturally present in vegetable foods. Previous findings showed that oral administration of Lactobacillus fermentum CRL1446 increased intestinal CE activity and improved oxidative status in mice. The aim of this work was to evaluate the in vitro CE activity of L. fermentum CRL1446 and the effect of bile on this activity, as well as strain resistance to simulated gastrointestinal tract (GIT) conditions and its ability to adhere to intestinal epithelium and influence its basal CE activity. L. fermentum CRL1446 and L. fermentum ATCC14932 (positive control for CE activity) were able to hydrolyse different synthetic hydroxycinnamates, with higher specificity toward methyl ferulate (3,853.73 and 899.19 U/g, respectively). Feruloyl esterase (FE) activity was mainly intracellular in L. fermentum CRL1446 and cell-surface associated in L. fermentum ATCC14932. Both strains tolerated simulated GIT conditions and were able to adhere ex vivo to intestinal epithelium. Pre-incubation of L. fermentum strains with bile increased FE activity in both whole cells and supernatants (~2-fold), compared to controls, suggesting that cells were permeabilised by bile, allowing more substrate to enter the cell and/or leakage of FE enzymes. Three-fold higher FE activities were detected in intestinal tissue fragments with adhered L. fermentum CRL1446 cells compared to control fragments (without bacteria), indicating that this strain provides exogenous FE activity and could stimulate esterase activity in the intestinal mucosa. Finally, we found that milk fat had a negative effect on FE activity of intestinal tissue, in absence or presence of adhered L. fermentum. These results help explaining the increase in intestinal FE activity previously observed in mice fed with L. fermentum CRL1446, and support the potential use of this strain for the development of new functional foods directed to oxidative stress-related ailments.
authors.affiliation: 1 Centro de Referencia para Lactobacilos (CERELA)-CONICET, Chacabuco 145, 4000 Tucumán, Argentina.|2 Facultad de Ciencias de la Salud, Universidad del Norte Santo Tomás de Aquino, 9 de Julio 165, 4000 Tucumán, Argentina.
authors.forename: M C Abeijón
authors.initials: MC
authors.lastname: Mukdsi
authors.affiliation: 3 Universidad Nacional de Tucumán, Ayacucho 491, 4000 Tucumán, Argentina.
authors.forename: E
authors.initials: E
authors.lastname: Argañaraz Martínez
authors.affiliation: 1 Centro de Referencia para Lactobacilos (CERELA)-CONICET, Chacabuco 145, 4000 Tucumán, Argentina.|3 Universidad Nacional de Tucumán, Ayacucho 491, 4000 Tucumán, Argentina.
authors.forename: A Perez
authors.initials: AP
authors.lastname: Chaia
authors.affiliation: 1 Centro de Referencia para Lactobacilos (CERELA)-CONICET, Chacabuco 145, 4000 Tucumán, Argentina.|2 Facultad de Ciencias de la Salud, Universidad del Norte Santo Tomás de Aquino, 9 de Julio 165, 4000 Tucumán, Argentina.|3 Universidad Nacional de Tucumán, Ayacucho 491, 4000 Tucumán, Argentina.
authors.forename: R B
authors.initials: RB
authors.lastname: Medina
mesh_terms: D000818:Animals; D001422:Bacterial Adhesion; D001646:Bile / Q000378:metabolism*; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D005750:Gastric Juice; D006017:Glycolipids / Q000378:metabolism; D006041:Goats; D007413:Intestinal Mucosa / Q000382:microbiology; D052197:Limosilactobacillus fermentum / Q000201:enzymology*; D008297:Male; D051379:Mice; D008892:Milk / Q000378:metabolism / Q000382:microbiology*
chemical_list: D006017:Glycolipids; D002265:Carboxylic Ester Hydrolases; C070196:feruloyl esterase
keywords: Lactobacillus fermentum; antioxidant hydroxycinnamic acids; functional foods; intestinal esterase activity; probiotics