Title Utjecaj glijadinskih frakcija na rast probiotičkih bakterija iz roda Lactobacillus u in vitro uvjetima Title (english) Effect of gliadin fractions on growth of probiotic bacteria genus Lactobacillus in vitro Author Marina Ožbolt Mentor Roberto Antolović (mentor)Committee member Roberto Antolović (predsjednik povjerenstva)Committee member Sandra Kraljević Pavelić (član povjerenstva)Committee member Mladenka Malenica Staver (član povjerenstva)Granter University of Rijeka Defense date and country 2016-09-16, Croatia Scientific / art field, BIOTECHNICAL SCIENCES Abstract Sastav humanog mikrobioma odraz je zdravlja pojedinca. Dokazano je da se kod ljudi s alergijama na hranu u njihovom mikrobiomu javlja neravnoteža između korisnih i štetnih bakterija. Probiotički pripravci se sve češće koriste ne samo u svrhu održavanja normalne mikroflore gastrointestinalnog sustava, već kao bioterapeutici u prevenciji i liječenju bolesti. Nepotpuna razgradnja glijadina u gastrointestinalnom traktu dovodi do pojave toksičnih peptida koji mogu aktivirati imunosni odgovor organizma, međutim, dokazano je da upravo probiotički pripravci mogu pomoći u sprječavanju njihovih štetnih učinaka. Kao potencijalna terapija posebice se ističu probiotički pripravci koji sadrže bakterije roda Lactobacillus. U ovom radu je istražen utjecaj glijadina na proliferaciju sedam Lactobacillus sojeva proučavanjem dvaju parametara – brzine rasta i duljine trajanja njihove lag faze u in vitro uvjetima. Bilo je potrebno izabrati najprikladnije otapalo za otapanje glijadina, a rezultati su pokazali da glijadin ima najveću topljivost u otopini 2% natrijeva klorida pH 6,2, a zatim u otopini 70% etanola u 50 mM Tris/HCl pH 7,2. Najmanje se otopio u otopini 70% etanola pH 6,6. Lactobacillus sojevi tretirani su glijadinom otopljenim u 70%-tnoj otopini etanola u 50 mM Tris/HCl te glijadinom otopljenim u otopini 2% natrijeva klorida. Rezultati pokazuju da glijadin u otopini 70% etanola u 50 mM Tris/HCl potiče proliferaciju nekih ispitivanih bakterija (L. helveticus XA/2, L. plantarum IIA/5, L. rhamnosus GG i L. paracasei spp. paracasei 1753), dok nema značajnog utjecaja na proliferaciju ostalih ispitivanih bakterija (L. acidophilus 145, L. casei VIIB/6, L. paracasei VIIB5). Rezultati pokusa s glijadinom otopljenim u otopini 2% natrijeva klorida, pokazuju da glijadin potiče proliferaciju bakterija L. helveticus XA/2 i L. plantarum IIA/5, dok nema značajnog utjecaja na proliferaciju ostalih ispitivanih bakterija (L. acidophilus 145, L. rhamnosus GG, L. casei VIIB/6, L. paracasei VIIB5, L. paracasei spp. paracasei 1753). Brzina rasta L. helveticus XA/2 i L. plantarum IIA/5 rasla je s porastom koncentracije glijadina u otopini 2% NaCl-a, no produljivala se i lag faza jer je navedenim bakterijama bilo potrebno dulje vrijeme za prilagodbu na nove uvjete.
Abstract (english) The composition of the human microbiome significantly influences the human health. It has been shown that imbalance between good (useful) and harmful bacteria is common in people who are suffering from food allergies. For that reason, use of probiotic supplements is increasing and they are broadly used not only to maintain the normal flora of the gastrointestinal system, but also as biotherapeutics in prevention and treatment of different gastrointestinal disorders. Incomplete digestion of gliadin in the gastrointestinal tract leads to formation of toxic peptides which can cause immune response in individuals. However, studies have shown that administration of probiotic supplements may prevent or mitigate adverse effects of these peptides. Among probiotic supplements, ones containing probiotic bacteria of genus Lactobacillus are standing out a potential therapy. This study shows effects of gliadin on proliferation of seven Lactobacillus strains, in vitro, studying two parameters - the growth rate and the lag phase. It was necessary to choose the most suitable solvent for dissolving the gliadin and the results showed that gliadin has the highest solubility in a solution of 2% sodium chloride pH 6,2, and then in a solution of 70 % ethanol in 50 mM Tris/HCl pH 7,2. Gliadine was least dissolved in 70 % ethanol solution pH 6,6. Lactobacillus strains were treated with gliadin disolved in solution of 70% ethanol in 50 mM Tris/HCl and with gliadin disolved in solution of 2% sodium chloridewhich was use din two different experiments. Results show that gliadin in solution of 70% ethanol in 50 mM Tris/HCl induced proliferation of tested bacteria (L. helveticus XA/2, L. plantarum II/5, L. rhamnosus GG and L. paracasei spp. paracasei 1753) or it had no significant effect on growth of other tested bacteria (L. acidophilus 145, L. casei VIIB/6, L. paracasei VIIB5). Results of the experiment with gliadin dissolved in 2% sodium chloride show that gliadin stimulated proliferation of L. helveticus XA/2 and L. plantarum II/5, but didn't have a significant effect on growth of other tested bacteria (L. acidophilus 145, L. rhamnosus GG, L. casei VIIB/6, L. paracasei VIIB5, L. paracasei spp. paracasei 1753). The growth rate of L. helveticus XA/2 and L. plantarum IIA/5 was higher with increasing concentration of gliadin in solution of 2% NaCl, but the lag fase was also longer because the bacteria needed more time to adjust to new conditions.
Keywords
glijadin
bolesti gastrointestinalnog sustava
Lactobacillus
proliferacija
Keywords (english)
gliadin
disorders of the gastrointestinal tract
Lactobacillus
proliferation
Language croatian URN:NBN urn:nbn:hr:193:975433 Project Number: 13.11.1.2.03. Study programme Title: Biotechnology in medicine Type of resource Text File origin Born digital Access conditions Closed access Terms of use Created on 2016-11-02 11:23:40
