Browsing by Author "Gümüştop, İsmail"

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Comparative Genomics of Lentilactobacillus parabuchneri isolated from dairy, KEM complex, Makgeolli, and Saliva Microbiomes
(BMC, 2022) Gumustop, Ismail; Ortakci, Fatih; AGÜ, Yaşam ve Doğa Bilimleri Fakültesi, Biyomühendislik Bölümü; Ortakçı, Fatih; Gümüştop, İsmail
Background: Lentilactobacillus parabuchneri is of particular concern in fermented food bioprocessing due to causing unwanted gas formation, cracks, and off-flavor in fermented dairy foods. This species is also a known culprit of histamine poisonings because of decarboxylating histidine to histamine in ripening cheese. Twenty-eight genomes in NCBI GenBank were evaluated via comparative analysis to determine genomic diversity within this species and identify potential avenues for reducing health associated risks and economic losses in the food industry caused by these organisms. Result: Core genome-based phylogenetic analysis revealed four distinct major clades. Eight dairy isolates, two strains from an unknown source, and a saliva isolate formed the first clade. Three out of five strains clustered on clade 2 belonged to dairy, and the remaining two strains were isolated from the makgeolli and Korean effective microorganisms (KEM) complex. The third and fourth clade members were isolated from Tete de Moine and dairy-associated niches, respectively. Whole genome analysis on twenty-eight genomes showed similar to 40% of all CDS were conserved across entire strains proposing a considerable diversity among L. parabuchneri strains analyzed. After assigning CDS to their corresponding function, similar to 79% of all strains were predicted to carry putative intact prophages, and similar to 43% of the strains harbored at least one plasmid; however, all the strains were predicted to encode genomic island, insertion sequence, and CRISPR-Cas system. A type I-E CRISPR-Cas subgroup was identified in all the strains, with the exception of DSM15352, which carried a type II-A CRISPR-Cas system. Twenty strains were predicted to encode histidine decarboxylase gene cluster that belongs to not only dairy but also saliva, KEM complex, and unknown source. No bacteriocin-encoding gene(s) or antibiotic resistome was found in any of the L. parabuchneri strains screened. Conclusion: The findings of the present work provide in-depth knowledge of the genomics of L. parabuchneri by comparing twenty-eight genomes available to date. For example, the hdc gene cluster was generally reported in cheese isolates; however, our findings in the current work indicated that it could also be encoded in those strains isolated from saliva, KEM complex, and unknown source. We think prophages are critical mobile elements of L. parabuchneri genomes that could pave the way for developing novel tools to reduce the occurrence of this unwanted species in the food industry.
Comparative genomics of Leuconostoc lactis strains isolated from human gastrointestinal system and fermented foods microbiomes
(BMCCAMPUS, 4 CRINAN ST, LONDON N1 9XW, ENGLAND, 2022) Gümüştop, İsmail; Ortakçı, Fatih; AGÜ, Yaşam ve Doğa Bilimleri Fakültesi, Biyomühendislik Bölümü; Gümüştop, İsmail; Ortakçı, Fatih
Background: Leuconostoc lactis forms a crucial member of the genus Leuconostoc and has been widely used in the fermentation industry to convert raw material into acidified and flavored products in dairy and plant-based food systems. Since the ecological niches that strains of Ln. lactis being isolated from were truly diverse such as the human gut, dairy, and plant environments, comparative genome analysis studies are needed to better understand the strain differences from a metabolic adaptation point of view across diverse sources of origin. We compared eight Ln. lactis strains of 1.2.28, aa_0143, BIOML-A1, CBA3625, LN19, LN24, WIKIM21, and WiKim40 using bioinformatics to elucidate genomic level characteristics of each strain for better utilization of this species in a broad range of applications in food industry. Results: Phylogenomic analysis of twenty-nine Ln. lactis strains resulted in nine clades. Whole-genome sequence analysis was performed on eight Ln. lactis strains representing human gastrointestinal tract and fermented foods microbiomes. The findings of the present study are based on comparative genome analysis against the reference Ln. lactis CBA3625 genome. Overall, a similar to 41% of all CDS were conserved between all strains. When the coding sequences were assigned to a function, mobile genetic elements, mainly insertion sequences were carried by all eight strains. All strains except LN24 and WiKim40 harbor at least one intact putative prophage region, and two of the strains contained CRISPR-Cas system. All strains encoded Lactococcin 972 bacteriocin biosynthesis gene clusters except for CBA3625. Conclusions: The findings in the present study put forth new perspectives on genomics of Ln. lactis via complete genome sequence based comparative analysis and further determination of genomic characteristics. The outcomes of this work could potentially pave the way for developing elements for future strain engineering applications.
Evaluating the microbial growth kinetics and artificial gastric digestion survival of a novel Pichia kudriavzevii FOL-04
(TAGEM Journals, 2022) Gümüştop, İsmail; Ortakci, Fatih; 0000-0002-1452-1283; 0000-0003-1319-0854; AGÜ, Yaşam ve Doğa Bilimleri Fakültesi, Biyomühendislik Bölümü; Gümüştop, İsmail; Ortakci, Fatih
Present study aims to explore Pichia kudriavzevii FOL-04 (FOL-04)’s: i) survival against artificial gastric juice (AGJ) and artificial bile juice (ABJ), ii) growth kinetics in shake flask (SF) and fed-batch trials (FBT). Survival of FOL-04 as measured by relative cell density (RCD) against AGJ and ABJ was screened at four different pH-levels (control, 3, 2, 1.5) and ox-bile concentrations (control, 0.2%, 1%, 2%), respectively. Growth kinetics was calculated by periodic measurement of OD600 in SF (225 rpm, 30°C) or in FBT using exponential feeding regimen where pH, dissolved-oxygen and temperature were controlled at 5.5, 21%, and 30°C, respectively. The doubling-time, maximum specific growth rate, and final cell densities achieved for SF and FBT were 81.7min, 1.67, 11.79 and 170.4 min, 4.75, 37.95, respectively. RCDs calculated were similar for pH=3 and control vs both were significantly higher(p<0.05) than pH=1.5 and 2 with the latter two pH-levels were not significantly different(p>0.05). RCDs were similar across control, 0.2%, and 1% ox-bile levels(p>0.05). However, 2% ox-bile yielded significantly lower RCD (p<0.05) compared to all except 1%. FOL-04 is a potential probiotic candidate showing robustness against AGJ and ABJ and remarkable biomass increase was achieved when grown under FBT which could pave the way for developing a yeast-based probiotic using this strain.
Next generation sequencing of a novel Loigolactobacillus coryniformis FOL-19 isolated from cheese and comparative genomic analysis with other L. coryniformis strains
(Abdullah Gül Üniversitesi, Fen Bilimleri Enstitüsü, 2023) Gümüştop, İsmail; AGÜ, Fen Bilimleri Enstitüsü, Biyomühendislik Ana Bilim Dalı
Loigolactobacillus coryniformis is a member of lactic acid bacteria isolated from various ecological niches. We isolated a novel L. coryniformis strain FOL-19 from artisanal Tulum cheese and performed the whole-genome sequencing for FOL-19 using Illumina NextSeq. Then, genomic characterization of FOL-19 against ten available whole genome sequences of the same species isolated from kimchi, silage, fermented meat, air of cowshed, and dairy was performed. The average genome size of 2.93 ±0.1 Mb, GC content of 42.96% ±0.002, number of CDS of 2905 ±165, number of tRNA of 56 ±10, and number of CRISPR elements of 6.55 ±1.83 was achieved. Both Type I and II Cas clusters were observed in L. coryniformis. Only one strain (CECT 5711) was predicted to encode a Carnocin CP52 bacteriocin gene cluster. The presence of CRISPR elements and Cas clusters suggests that L. coryniformis holds a promising potential for being a reservoir for new CRISPR-based tools. These findings put a step forward for the genomic characterization of L. coryniformis strains for biotechnological applications via genome-guided strain selection to identify industrially relevant traits.