In vitro adhesion of the S-layer expressing Lactobacillus strains to the proteins of the extracellular matrix

Cilj ovog rada je ispitati in vitro adheziju pojedinih Lactobacillus sojeva, koji eksprimiraju S-layer površinski protein, na proteine ekstracelularnog matriksa. Eksperimenti su provedeni s četiri različita Lactobacillus soja: Lactobacillus brevis ZG1-K7, Lactobacillus brevis SF9B, Lactobacillus paraplantarum SF15B i Lactobacillus plantarum D6, iz Zbirke bakterija mliječne kiseline Laboratorija za tehnologiju antibiotika, enzima, probiotika i starter kultura, Prehrambeno-biotehnološkog fakulteta, koja sadrži više od 200 različitih sojeva bakterija mliječne kiseline. SDS-PAGE analizom površinskih proteina ovih Lactobacillus sojeva ustanovljeno je da eksprimiraju S-layer površinske proteine. Sposobnost adhezije Lactobacillus sojeva se međusobno razlikuje ovisno o soju i specifična je ovisno o proteinu ekstracelularnog matriksa. Uklanjanje intaktnog S-layer proteina s površine bakterijske stanice, ekstrakcijom s gvanidin hidrokloridom, uzrokovalo je smanjenje adhezije kod svih Lactobacillus sojeva na fibronektin i kod većine sojeva na kolagen i laminin, dok je adhezija na laminin bila povećana, ali neznatno kod sojeva L. brevis ZG1-K7 i L. paraplantarum SF15B, te na kolagen kod sojeva L. brevis SF9B i L. paraplantarum SF15B. Prema rezultatima odabrani Lactobacillus sojevi se uspješno adheziraju na proteine ECM, što se značajno smanji nakon tretmana bakterijskih stanica s proteinazom K.The aim of this work was to characterize ability of several S-layer-expressing Lactobacillus to adhere to the main proteins of the extracellular matrix (ECM). Four different Lactobacillus strains from the Collection of Lactic Acid Bacteria (ZBMK in Laboratory of Antibiotic, Enzyme, Probiotic and Starter Cultures Technology, Faculty of Food Technology and Biotechnology, over 200 strains), were used: Lactobacillus brevis ZG1-K7, Lactobacillus brevis SF9B; Lactobacillus paraplantarum SF15B and Lactobacillus plantarum D6. Four Lactobacillus pure cultures exhibited S-layer proteins according to the SDS-PAGE analyses. The binding capability of the Lactobacillus strains varied greatly among the isolates and was specific depending of ECM protein. Removal of the intact S-layer proteins from the bacterial surface by extraction with guanidine hydrochloride has reduced the adhesion of all of Lactobacillus strains to fibronectin and in most strains to laminin and collagen; whereas, the adhesiveness to laminin was increased, althought slightly, with L. brevis ZG1-K7 and L. paraplantarum SF15B, and to collagen with L. brevis SF9B and L. paraplantarum SF15B. Binding tests showed that mostly selected Lactobacillus strains adhere to ECM proteins in a proteinase K-sensitive manner