Inhibition of enteropathogens adhesion to human enterocyte-like HT-29 cells by a dairy strain of Propionibacterium acidipropionici.

Adhesion to the host intestinal mucosa is considered relevant for orally delivered probiotics as it prolongs their persistence in the gut and their health promoting effects. Classical propionibacteria are microorganisms of interest due to their role as dairy starters as well as for their functions as probiotics. Propionibacterium acidipropionici Q4, is a dairy strain isolated from a Swiss-type cheese made in Argentina that displays probiotic potential. In the present work we assessed the ability of this strain to adhere to the human enterocyte-like HT-29 cell line and to counteract the adhesion of two common human enteropathogens, such as Escherichia coli C3 and Salmonella Enteritidis 90/390. The results were compared with those obtained with the well-known probiotic Lactobacillus rhamnosus GG. P. acidipropionici Q4 showed a high adhesion capacity, even higher than the reference strain L. rhamnosus GG (42.3±4.4% and 36.2±2.3%, respectively), whereas adhesion of enteropathogens was significantly lower (25.2±2.2% for E. coli and 21.0±3.4% for S. Enteritidis). Propionibacteria as well as lactobacilli were able to inhibit by exclusion and competition the adherence of E. coli C3 and S. Enteritidis 90/390 whereas only L. rhamnosus GG displaced S. Enteritidis from HT-29 intestinal cells. Inhibition of pathogens by propionibacteria was not exerted by antimicrobials or coaggregation but was mainly due to exclusion by cell surface components, such as proteins and carbohydrates. The relevance of cell surface proteins (CSP) for preventing pathogens infection was confirmed by their concentration dependent effect observed for both pathogens: 100 µg/ml of CSP inhibited E. coli attachment almost as untreated propionibacteria, whereas it partially inhibited the attachment of S. Enteritidis. Results suggest that P. acidipropionici Q4 could be considered for the development of propionibacteria containing functional foods helpful in counteracting enteropathogen infection.

The effect of processing conditions on the stability of fructooligosaccharides in acidic food products.

The effect of processing conditions (temperature and degree of polymerisation, DP) on the stability of short-chain fructooligosaccharides (sc-FOS) was investigated in three reaction media (sodium citrate buffer and orange and tomato juices) in a kinetic study at pH 3.5. In addition, kinetic equations as a function of temperature and pH were developed, using published data. Pentasaccharides were more stable to heat treatment than were trisaccharides under all of the conditions tested. In addition, the sc-FOS were more stable in orange juice, followed by tomato juice and citrate buffer. The results showed that, in addition to temperature and pH, the DP and food matrix, including the type of pasteurisation, must be considered when processing foods enriched with sc-FOS. Furthermore, the continuous thermal processing simulation for each of the equivalent processes at 90 °C revealed that the percent retention of sc-FOS is greater than 95% at temperatures above 95 °C.

Assessment of antiproliferative activity of pectic substances obtained by different extraction methods from rapeseed cake on cancer cell lines.

In this work the antiproliferative activity of pectic substances obtained by different extraction methods from defatted rapeseed cake was assessed on cancer cell lines. The process consisted of sequential treatment with alkalized water (pH∼8), EDTA (0.01 M), alkaline protease (Alkalase 2.4L) and a commercial pectinase preparation (Viscozyme L or Pectinex Ultra SP-L). Pectic extracts identification was performed using spectroscopy and chromatography techniques. FT-IR and HPLC-IR results suggest that the neutral pectic extracts produced would be arabinogalactans and ß-galactans. All the pectic substances extracted (acid and neutral) from RSC exhibited antiproliferative activity, being more effective on MCF-7 cells than Caco-2. The most effective pectic extract was obtained by Alkalase 2.4 L which killed over 80% of MCF-7 cells and 60% of Caco-2 cells. At less than 10 mg/mL pectic extracts enriched in neutral sugars also exhibited antiproliferative activity (50 and 40%, respectively), which was superior to the modified citric pectins activity at the same concentration for the breast cancer cell line (61.6% for MCF-7 and 49.9% for Caco-2 cells). These results show that the antiproliferative activity depends on both the type of pectin (acid or neutral) and the extraction procedure.

Enzymatic synthesis of fructooligosaccharides with high 1-kestose concentrations using response surface methodology.

Response surface methodology was used as an optimization tool for the production of short chain fructooligosaccharides (sc-FOS) using the commercial cellulolytic enzyme preparation, Rohapect CM. Three independent variables, temperature, concentrations of sucrose and enzyme were tested in the reaction medium. The responses of the design were, yield (gsc-FOS/100 g initial sucrose), 1-kestose (g/100 g sc-FOS) and volumetric productivity (gsc-FOS/Lh). Significant effects on the three responses included a quadratic effect (temperature), a linear effect (sucrose and enzyme concentrations) and an interaction between temperature and sucrose concentration. The cost-effective conditions to support the process in a high competitive market were 50 °C, 6.6 TU/mL enzyme, 2.103 M sucrose in 50 mM acetate buffer at pH 5.5, and the synthesis for a 5 h reaction time. Under these conditions, a high YP/S (63.8%), QP (91.9 g/Lh) and sGF2 (68.2%) was achieved.