Isolation and molecular identification of industrially important enzyme producer yeasts from tree barks and fruits.

This study aimed to identify the yeast strains associated with the tree bark samples collected from the Aegean and Marmara regions and from rotten fruit samples. Fifty-one yeast strains were successfully isolated and screened for their abilities to produce industrially important extracellular enzymes. Thirty isolates demonstrated ability to produce at least two different enzymes and were selected for subsequent molecular identification using sequence analysis of ITS region and D1/D2 domain of the 26S rDNA. The most prevalent strains belonged to Papiliotrema laurentii (%23), Papiliotrema terrestris (%13) and Candida membranifaciens (%10). Papiliotrema laurentii and Papiliotrema terrestris recorded the highest enzymatic activities for all the screened enzymes. To the best of our knowledge, this is the first report that identifies the yeast strains associated with the tree barks of Turkey and among the limited comprehensive studies that screened considerable number of isolates for their ability to produce several industrially important enzymes.

Inhibition of Salmonella enteritidis biofilms by Salmonella invasion protein-targeting aptamer.

The current study aimed to assess the inhibitory effect of a DNA aptamer (Apt17) which targeted Salmonella invasion proteinA (SipA). The effect of Apt17, on biofilm formation by two Salmonella enteritidis strains, was tested either separately or in combination with ampicillin at different Sub MIC concentrations. Maximum inhibitory effect equivalent to 24.34% and 26.81% was recorded when Apt17 was co-incubated with S. enteritidis TM 6 and S. enteritidis TM 68 respectively for 13 h. The inhibitory effect of Apt17 was also confirmed with Triphenyl Tetrazolium Chloride. Under Scanning Electron Microscope, the presence of Apt17 resulted in altered three dimensional structure. While the treated cells of S. enteritidis TM 6 were arranged as monolayers, the sessile aggregates of S. enteritidis TM 68 appeared thinner and exhibited less surface coverage when compared to control. Moreover, the treated cells lost their exopolysaccharide matrix. The co-incubation of Apt17 with ampicillin MIC/10 for 24 h, inhibited the biofilms of S. enteritidis TM 6 and S. enteritidis TM 68 by 12.5 and 20.9% respectively. This study demonstrated quantitative and qualitative antibiofilm effect of Apt17 against the biofilms of two Salmonella enteritidis strains. According to our knowledge, this is the first study employing an aptamer that targets SipA protein to inhibit biofilm formation in Salmonella.

The effect of carbon, nitrogen and iron ions on mono-rhamnolipid production and rhamnolipid synthesis gene expression by Pseudomonas aeruginosa ATCC 15442.

Pseudomonas spp. are the main producers of rhamnolipids. These products have applications in pharmaceuticals, cosmetics, food industry and bioremediation. The biosynthesis of rhamnolipids is influenced by nutrient composition, pH and temperature. In this study, the impact of nutrients on the expression levels of rhamnolipid synthesis genes was evaluated in P. aeruginosa ATCC 15442. Glucose and glycerol were used as carbon sources; while, NaNO3, NH4NO3 and yeast extract/peptone were employed as nitrogen sources. The effect of different concentrations of Fe2+ and Fe3+ on rhamnolipid synthesis genes was also evaluated. Highest biosurfactant production was obtained in minimal medium supplemented with glucose, NaNO3 and Fe2+. Two rhamnolipid synthesis genes, rhlA and rhlB, were amplified with PCR. CapLC ESI-Ion trap-MS/MS detected only mono-rhamnolipid Rha-C10-C10 in the extract. Although similar induction levels were recorded in the presence of 0.05 g/L iron ions, the presence of Fe2+ resulted in higher expression levels than Fe3+ at concentrations equivalent to 0.025 and 0.075 g/L.

Single-stranded DNA (ssDNA) Aptamer targeting SipA protein inhibits Salmonella Enteritidis invasion of intestinal epithelial cells.

Salmonella Enteritidis is an important pathogen that can invade the intestinal cells of its host causing salmonellosis. SipA protein, an effector protein secreted by T3SS, maintains invasion of host cells more efficient. Thus, inhibitory aptamers against SipA protein were developed using magnetic bead-based Systematic Evolution of Ligands by Exponential Enrichment (SELEX) method. The enriched sequences were obtained after 9 SELEX rounds. Among which, an aptamer namely Apt17 displayed Kd values equivalent to 114.9 and 63.4 nM at 27 °C and 37 °C, respectively. The effect of Apt17 on adhesion and invasion of Caco-2 cells by the tested strains was determined. While the adhesion and invasion of Salmonella Enteritidis TM 6 were inhibited by 70% and 37.7%, those of Salmonella Enteritidis TM 68 were inhibited by 45.71% and 39.5% respectively. These results represent a corner stone for future studies that could aim to develop putative inhibitors against Salmonellosis.

Molecular characterization and lipase profiling of the yeasts isolated from environments contaminated with petroleum.

In the present study, 120 yeast isolates from different sources (active sludge, soil, and wastewater samples obtained from petroleum refinery and soil contaminated by petroleum) were obtained. The yeast isolates were screened for lipase production and twelve of the isolates (D3, D17, D24, D27, D30, D38, D40, D42, D44, D46, D56, and D57) exhibited lipase activity. Molecular characterization of the yeasts showing the lipase production was performed with RFLP of ITS1-5.8S-ITS2 and 18S rRNA and sequence analysis of D1/D2 domain of 26S rRNA. The 26S rRNA sequencing revealed that four new strains, D38, D40, D44 and D57 identified as Rhodotorula slooffiae, Candida davisiana, Cryptococcus diffluens, and Cryptococcus uzbekistanensis, respectively, are lipase producing yeast species. This study is the first report showed lipase production by these species. The other lipase producing strains identified as Candida parapsilosis (D3), Rhodotorula muciloginosa (D17 and D42), Cryptococcus albidus (D24, D27, D30, and D56), and Wickerhamomyces anomalus (D46).