Miljković, Marija

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The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties

Malešević, Milka; Stanisavljević, Nemanja; Miljković, Marija; Jovčić, Branko; Filipić, Brankica; Studholme, David; Kojić, Milan

(Elsevier B.V., 2021)

TY  - JOUR
AU  - Malešević, Milka
AU  - Stanisavljević, Nemanja
AU  - Miljković, Marija
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Studholme, David
AU  - Kojić, Milan
PY  - 2021
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3729
AB  - Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis bv. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid pS74 is EPS production, while plasmid pS127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid pS127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.
PB  - Elsevier B.V.
T2  - International Journal of Food Microbiology
T1  - The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties
VL  - 337
DO  - 10.1016/j.ijfoodmicro.2020.108935
ER  - 
@article{
author = "Malešević, Milka and Stanisavljević, Nemanja and Miljković, Marija and Jovčić, Branko and Filipić, Brankica and Studholme, David and Kojić, Milan",
year = "2021",
abstract = "Plasmids are autonomous episomally replicating genetic elements, which carry backbone genes important for the replication and maintenance within their host, and accessory genes that might confer an advantage to their host under specific selective pressure in its ecological niche. The genome of dairy isolate L. lactis subsp. lactis bv. diacetylactis S50 was sequenced using the PacBio SMRT Cell Seq-RSII platform and revealed to possess one of the largest plasmidomes among L. lactis strains studied so far, harboring six plasmids: pS6 (5553 bp), pS7a (7308 bp), pS7b (7266 bp), pS19 (19,027 bp), pS74 (74,256 bp) and pS127 (127,002 bp) in total representing 8.9% of genome size (240,412 bp). Based on predicted plasmid replication proteins and origins it appears that all six plasmids replicate via the theta-type mechanism. The two the largest plasmids (pS74 and pS127), carry a number of genes known to be important for growth and survival in the dairy environment. These genes encode technological functions such as bacteriocin production, protein degradation, magnesium and cobalt/nickel transporters, selenium binding, exopolysaccharides (EPS) production, bacteriophage and stress resistance. Beside genes for replication, the small plasmids (pS6, pS7a, pS7a, and pS19) also carry genes important for mobilization and host survival such as type I restriction-modification (R-M) system, metal transporters, enzymes and transcriptional regulators. All plasmids in S50 strain are mobilizable, containing an oriT sequences, while pS127 is self-conjugative and allows for mobilization of the other plasmids. Small plasmids are prone to structural and segregational instability, while pS127 appeared to be segregationally stable thanks to the possession of two partition systems. The main characteristic of plasmid pS74 is EPS production, while plasmid pS127 is characterized by proteinase and multiple bacteriocins, tra locus, phage abortive systems and metal transporters. In addition to LcnA and LcnB, plasmid pS127 encodes several bacteriocin-pheromone molecules and a new bacteriocin named LcnS50, with narrow spectrum of action limited to lactococci, that has been successfully cloned and heterologously expressed.",
publisher = "Elsevier B.V.",
journal = "International Journal of Food Microbiology",
title = "The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties",
volume = "337",
doi = "10.1016/j.ijfoodmicro.2020.108935"
}
Malešević, M., Stanisavljević, N., Miljković, M., Jovčić, B., Filipić, B., Studholme, D.,& Kojić, M.. (2021). The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology
Elsevier B.V.., 337.
https://doi.org/10.1016/j.ijfoodmicro.2020.108935
Malešević M, Stanisavljević N, Miljković M, Jovčić B, Filipić B, Studholme D, Kojić M. The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties. in International Journal of Food Microbiology. 2021;337.
doi:10.1016/j.ijfoodmicro.2020.108935 .
Malešević, Milka, Stanisavljević, Nemanja, Miljković, Marija, Jovčić, Branko, Filipić, Brankica, Studholme, David, Kojić, Milan, "The large plasmidome of Lactococcus lactis subsp. lactis bv. diacetylactis S50 confers its biotechnological properties" in International Journal of Food Microbiology, 337 (2021),
https://doi.org/10.1016/j.ijfoodmicro.2020.108935 . .
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Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials

Miljković, Marija; Jovanović, Sofija; O'Connor, Paula M.; Mirković, Nemanja; Jovčić, Branko; Filipić, Brankica; Dinić, Miroslav; Studholme, David John; Fira, Đorđe; Cotter, Paul D.; Kojić, Milan

(Public Library Science, San Francisco, 2019)

TY  - JOUR
AU  - Miljković, Marija
AU  - Jovanović, Sofija
AU  - O'Connor, Paula M.
AU  - Mirković, Nemanja
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Dinić, Miroslav
AU  - Studholme, David John
AU  - Fira, Đorđe
AU  - Cotter, Paul D.
AU  - Kojić, Milan
PY  - 2019
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3372
AB  - Bacteria active against multi-drug resistant pathogens, isolated by direct selection of colonies from clover silage samples, produce zones of inhibition against two Gram-negative (Klebsiella pneumoniae Ni9 and Pseudomonas aeruginosa MMA83) and two Gram-positive (Staphylococcus aureus ATCC25923 and Listeria monocytogenes ATCC19111) pathogens. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 produced the largest zones of inhibition against all four pathogens when grown in LB broth with aeration at 37 degrees C. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 were identified as Brevibacillus laterosporus and pulsed field gel electrophoresis and extracellular protein profiles showed that three different strains (BGSP7, BGSP9 and BGSP11) were isolated. A semi-native SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) gel overlay assay showed that BGSP7 and BGSP9 produce small antimicrobial molecules of about 1.5 kDa, while BGSP11 produces antimicrobial molecules of 1.5 and 6 kDa active against S. aureus ATCC25923. Amino acid analysis of two antimicrobial molecules (1583.73 Da; from BGSP7 and 1556.31 Da; from BGSP11) revealed that they have a similar composition and differ only by virtue of the presence of a methionine which is present only in BGSP11 molecule. Genome sequencing of the three isolates revealed the presence of gene clusters associated with the production of non-ribosomally synthesized peptides (brevibacillin, bogorol, gramicidin S, plipastatin and tyrocin) and bacteriocins (laterosporulin, a lactococcin 972-like bacteriocin, as well as putative linocin M18, sactipeptide, UviB and lantipeptide-like molecules). Ultimately, the purification of a number of antimicrobial molecules from each isolate suggests that they can be considered as potent biocontrol strains that produce an arsenal of antimicrobial molecules active against Gram-positive and Gram-negative multi-resistant pathogens, fungi and insects.
PB  - Public Library Science, San Francisco
T2  - PLoS One
T1  - Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials
VL  - 14
IS  - 5
DO  - 10.1371/journal.pone.0216773
ER  - 
@article{
author = "Miljković, Marija and Jovanović, Sofija and O'Connor, Paula M. and Mirković, Nemanja and Jovčić, Branko and Filipić, Brankica and Dinić, Miroslav and Studholme, David John and Fira, Đorđe and Cotter, Paul D. and Kojić, Milan",
year = "2019",
abstract = "Bacteria active against multi-drug resistant pathogens, isolated by direct selection of colonies from clover silage samples, produce zones of inhibition against two Gram-negative (Klebsiella pneumoniae Ni9 and Pseudomonas aeruginosa MMA83) and two Gram-positive (Staphylococcus aureus ATCC25923 and Listeria monocytogenes ATCC19111) pathogens. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 produced the largest zones of inhibition against all four pathogens when grown in LB broth with aeration at 37 degrees C. Isolates BGSP7, BGSP9, BGSP11 and BGSP12 were identified as Brevibacillus laterosporus and pulsed field gel electrophoresis and extracellular protein profiles showed that three different strains (BGSP7, BGSP9 and BGSP11) were isolated. A semi-native SDS-PAGE (sodium dodecyl sulphate-polyacrylamide gel electrophoresis) gel overlay assay showed that BGSP7 and BGSP9 produce small antimicrobial molecules of about 1.5 kDa, while BGSP11 produces antimicrobial molecules of 1.5 and 6 kDa active against S. aureus ATCC25923. Amino acid analysis of two antimicrobial molecules (1583.73 Da; from BGSP7 and 1556.31 Da; from BGSP11) revealed that they have a similar composition and differ only by virtue of the presence of a methionine which is present only in BGSP11 molecule. Genome sequencing of the three isolates revealed the presence of gene clusters associated with the production of non-ribosomally synthesized peptides (brevibacillin, bogorol, gramicidin S, plipastatin and tyrocin) and bacteriocins (laterosporulin, a lactococcin 972-like bacteriocin, as well as putative linocin M18, sactipeptide, UviB and lantipeptide-like molecules). Ultimately, the purification of a number of antimicrobial molecules from each isolate suggests that they can be considered as potent biocontrol strains that produce an arsenal of antimicrobial molecules active against Gram-positive and Gram-negative multi-resistant pathogens, fungi and insects.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One",
title = "Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials",
volume = "14",
number = "5",
doi = "10.1371/journal.pone.0216773"
}
Miljković, M., Jovanović, S., O'Connor, P. M., Mirković, N., Jovčić, B., Filipić, B., Dinić, M., Studholme, D. J., Fira, Đ., Cotter, P. D.,& Kojić, M.. (2019). Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials. in PLoS One
Public Library Science, San Francisco., 14(5).
https://doi.org/10.1371/journal.pone.0216773
Miljković M, Jovanović S, O'Connor PM, Mirković N, Jovčić B, Filipić B, Dinić M, Studholme DJ, Fira Đ, Cotter PD, Kojić M. Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials. in PLoS One. 2019;14(5).
doi:10.1371/journal.pone.0216773 .
Miljković, Marija, Jovanović, Sofija, O'Connor, Paula M., Mirković, Nemanja, Jovčić, Branko, Filipić, Brankica, Dinić, Miroslav, Studholme, David John, Fira, Đorđe, Cotter, Paul D., Kojić, Milan, "Brevibacillus laterosporus strains BGSP7, BGSP9 and BGSP11 isolated from silage produce broad spectrum multi-antimicrobials" in PLoS One, 14, no. 5 (2019),
https://doi.org/10.1371/journal.pone.0216773 . .
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PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.

Novović, Katarina; Malesević, Milka; Filipić, Brankica; Mirković, Nemanja; Miljković, Marija; Kojić, Milan; Jovčić, Branko

(Springer, New York, 2019)

TY  - JOUR
AU  - Novović, Katarina
AU  - Malesević, Milka
AU  - Filipić, Brankica
AU  - Mirković, Nemanja
AU  - Miljković, Marija
AU  - Kojić, Milan
AU  - Jovčić, Branko
PY  - 2019
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3313
AB  - Pseudomonas aeruginosa, which is a clinically important representative of Pseudomonas spp., has been recognized as causative agent of severe nosocomial infections worldwide. An increase in antibiotic resistance of P. aeruginosa clinical strains could be attributed to their capacity to acquire resistance through mobile genetic elements such as mobile integrons that are present in one-half of multidrug-resistant P. aeruginosa strains. Mobile class 1 integrons are recognized as genetic elements involved in the rapid dissemination of multiple genes encoding for antibiotic resistance. The LexA protein is a major repressor of integrase transcription, but differences in transcription regulation among bacterial species have also been noted. In this study, the promoter activity of class 1 integron integrase gene (intI1) and its variant lacking the LexA binding site in Pseudomonas putida WCS358 wild type, rpoS and psrA was analysed. The results show that the activity of the intI1 gene promoter decreased in the rpoS and psrA mutants in the stationary phase of growth compared to the wild type, which indicates the role of RpoS and PsrA proteins in the positive regulation of integrase transcription. Additionally, it was determined that the activity of the lexA gene promoter decreased in rpoS and psrA, and thus, we propose that PsrA indirectly regulates the intI1 gene promoter activity through regulation of lexA gene expression in co-operation with some additional regulators. In this study, intI1 gene expression was shown to be controlled by two major stress response (SOS and RpoS) regulons, which indicates that integrase has evolved to use both systems to sense the cell status.
PB  - Springer, New York
T2  - Current Microbiology
T1  - PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.
VL  - 76
IS  - 3
SP  - 320
EP  - 328
DO  - 10.1007/s00284-019-01626-7
ER  - 
@article{
author = "Novović, Katarina and Malesević, Milka and Filipić, Brankica and Mirković, Nemanja and Miljković, Marija and Kojić, Milan and Jovčić, Branko",
year = "2019",
abstract = "Pseudomonas aeruginosa, which is a clinically important representative of Pseudomonas spp., has been recognized as causative agent of severe nosocomial infections worldwide. An increase in antibiotic resistance of P. aeruginosa clinical strains could be attributed to their capacity to acquire resistance through mobile genetic elements such as mobile integrons that are present in one-half of multidrug-resistant P. aeruginosa strains. Mobile class 1 integrons are recognized as genetic elements involved in the rapid dissemination of multiple genes encoding for antibiotic resistance. The LexA protein is a major repressor of integrase transcription, but differences in transcription regulation among bacterial species have also been noted. In this study, the promoter activity of class 1 integron integrase gene (intI1) and its variant lacking the LexA binding site in Pseudomonas putida WCS358 wild type, rpoS and psrA was analysed. The results show that the activity of the intI1 gene promoter decreased in the rpoS and psrA mutants in the stationary phase of growth compared to the wild type, which indicates the role of RpoS and PsrA proteins in the positive regulation of integrase transcription. Additionally, it was determined that the activity of the lexA gene promoter decreased in rpoS and psrA, and thus, we propose that PsrA indirectly regulates the intI1 gene promoter activity through regulation of lexA gene expression in co-operation with some additional regulators. In this study, intI1 gene expression was shown to be controlled by two major stress response (SOS and RpoS) regulons, which indicates that integrase has evolved to use both systems to sense the cell status.",
publisher = "Springer, New York",
journal = "Current Microbiology",
title = "PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.",
volume = "76",
number = "3",
pages = "320-328",
doi = "10.1007/s00284-019-01626-7"
}
Novović, K., Malesević, M., Filipić, B., Mirković, N., Miljković, M., Kojić, M.,& Jovčić, B.. (2019). PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.. in Current Microbiology
Springer, New York., 76(3), 320-328.
https://doi.org/10.1007/s00284-019-01626-7
Novović K, Malesević M, Filipić B, Mirković N, Miljković M, Kojić M, Jovčić B. PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp.. in Current Microbiology. 2019;76(3):320-328.
doi:10.1007/s00284-019-01626-7 .
Novović, Katarina, Malesević, Milka, Filipić, Brankica, Mirković, Nemanja, Miljković, Marija, Kojić, Milan, Jovčić, Branko, "PsrA Regulator Connects Cell Physiology and Class 1 Integron Integrase Gene Expression Through the Regulation of lexA Gene Expression in Pseudomonas spp." in Current Microbiology, 76, no. 3 (2019):320-328,
https://doi.org/10.1007/s00284-019-01626-7 . .
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5

LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity

Miljković, Marija; Malesević, Milka; Filipić, Brankica; Vukotić, Goran; Kojić, Milan

(Hindawi Ltd, London, 2018)

TY  - JOUR
AU  - Miljković, Marija
AU  - Malesević, Milka
AU  - Filipić, Brankica
AU  - Vukotić, Goran
AU  - Kojić, Milan
PY  - 2018
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/3192
AB  - Restriction enzymes are the main defence system against foreign DNA, in charge of preserving genome integrity. Lactococcus raffinolactis BGTRK10-1 expresses LraI Type II restriction-modification enzyme, whose activity is similar to that shown for EcoRI; LraI methyltransferase protects DNA from EcoRI cleavage. The gene encoding LraI endonuclease was cloned and overexpressed in E. coli. Purified enzyme showed the highest specific activity at lower temperatures (between 13 degrees C and 37 degrees C) and was stable after storage at -20 degrees C in 50% glycerol. The concentration of monovalent ions in the reaction buffer required for optimal activity of LraI restriction enzyme was 100 mM or higher. The recognition and cleavage sequence for LraI restriction enzyme was determined as 5'-G/AATTC-3', indicating that LraI restriction enzyme is an isoschizomer of EcoRI. In the reaction buffer with a lower salt concentration, LraI exhibits star activity and specifically recognizes and cuts another alternative sequence 5'-A/AATTC-3', leaving the same sticky ends on fragments as EcoRI, which makes them clonable into a linearized vector. Phylogenetic analysis based on sequence alignment pointed out the common origin of LraI restriction-modification system with previously described EcoRI-like restriction-modification systems.
PB  - Hindawi Ltd, London
T2  - Biomed Research International
T1  - LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity
DO  - 10.1155/2018/5657085
ER  - 
@article{
author = "Miljković, Marija and Malesević, Milka and Filipić, Brankica and Vukotić, Goran and Kojić, Milan",
year = "2018",
abstract = "Restriction enzymes are the main defence system against foreign DNA, in charge of preserving genome integrity. Lactococcus raffinolactis BGTRK10-1 expresses LraI Type II restriction-modification enzyme, whose activity is similar to that shown for EcoRI; LraI methyltransferase protects DNA from EcoRI cleavage. The gene encoding LraI endonuclease was cloned and overexpressed in E. coli. Purified enzyme showed the highest specific activity at lower temperatures (between 13 degrees C and 37 degrees C) and was stable after storage at -20 degrees C in 50% glycerol. The concentration of monovalent ions in the reaction buffer required for optimal activity of LraI restriction enzyme was 100 mM or higher. The recognition and cleavage sequence for LraI restriction enzyme was determined as 5'-G/AATTC-3', indicating that LraI restriction enzyme is an isoschizomer of EcoRI. In the reaction buffer with a lower salt concentration, LraI exhibits star activity and specifically recognizes and cuts another alternative sequence 5'-A/AATTC-3', leaving the same sticky ends on fragments as EcoRI, which makes them clonable into a linearized vector. Phylogenetic analysis based on sequence alignment pointed out the common origin of LraI restriction-modification system with previously described EcoRI-like restriction-modification systems.",
publisher = "Hindawi Ltd, London",
journal = "Biomed Research International",
title = "LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity",
doi = "10.1155/2018/5657085"
}
Miljković, M., Malesević, M., Filipić, B., Vukotić, G.,& Kojić, M.. (2018). LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity. in Biomed Research International
Hindawi Ltd, London..
https://doi.org/10.1155/2018/5657085
Miljković M, Malesević M, Filipić B, Vukotić G, Kojić M. LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity. in Biomed Research International. 2018;.
doi:10.1155/2018/5657085 .
Miljković, Marija, Malesević, Milka, Filipić, Brankica, Vukotić, Goran, Kojić, Milan, "LraI from Lactococcus raffinolactis BGTRK10-1, an Isoschizomer of EcoRI, Exhibits Ion Concentration-Dependent Specific Star Activity" in Biomed Research International (2018),
https://doi.org/10.1155/2018/5657085 . .
4
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Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?

Terzić-Vidojević, Amarela; Veljović, Katarina; Begović, Jelena; Filipić, Brankica; Popović, Dušanka; Tolinacki, Maja; Miljković, Marija; Kojić, Milan; Golić, Nataša

(Frontiers Media Sa, Lausanne, 2015)

TY  - JOUR
AU  - Terzić-Vidojević, Amarela
AU  - Veljović, Katarina
AU  - Begović, Jelena
AU  - Filipić, Brankica
AU  - Popović, Dušanka
AU  - Tolinacki, Maja
AU  - Miljković, Marija
AU  - Kojić, Milan
AU  - Golić, Nataša
PY  - 2015
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2320
AB  - Enterococci represent the most controversial group of dairy bacteria. They are found to be the main constituent of many traditional Mediterranean dairy products and contribute to their characteristic taste and flavor. On the other hand, during the last 50 years antibiotic resistant enterococci have emerged as leading causes of nosocomial infections worldwide. The aim of this study was to determine the diversity, technological properties, antibiotic susceptibility and virulence traits of 636 enterococci previously isolated from 55 artisan dairy products from 12 locations in the Western Balkan countries (WBC) of Serbia, Croatia and Bosnia and Herzegovina. All strains were identified both by microbiological and molecular methods. The predominant species was Enterococcus durans, followed by Enterococcus faecalis and Enterococcus faecium. Over 44% of the isolates were resistant to ciprofloxacin and erythromycin, while 26.2% of the isolates were multi resistant to three or more antibiotics belonging to different families. 185 isolates (29.1%) were susceptible to all 13 of the antibiotics tested. The antibiotic-susceptible isolates were further tested for possible virulence genes and the production of biogenic amines. Finally, five enterococci isolates were found to be antibiotic susceptible with good technological characteristics and without virulence traits or the ability to produce biogenic amines, making them possible candidates for biotechnological application as starter cultures in the dairy industry.
PB  - Frontiers Media Sa, Lausanne
T2  - Frontiers in Microbiology
T1  - Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?
VL  - 6
DO  - 10.3389/fmicb.2015.00954
ER  - 
@article{
author = "Terzić-Vidojević, Amarela and Veljović, Katarina and Begović, Jelena and Filipić, Brankica and Popović, Dušanka and Tolinacki, Maja and Miljković, Marija and Kojić, Milan and Golić, Nataša",
year = "2015",
abstract = "Enterococci represent the most controversial group of dairy bacteria. They are found to be the main constituent of many traditional Mediterranean dairy products and contribute to their characteristic taste and flavor. On the other hand, during the last 50 years antibiotic resistant enterococci have emerged as leading causes of nosocomial infections worldwide. The aim of this study was to determine the diversity, technological properties, antibiotic susceptibility and virulence traits of 636 enterococci previously isolated from 55 artisan dairy products from 12 locations in the Western Balkan countries (WBC) of Serbia, Croatia and Bosnia and Herzegovina. All strains were identified both by microbiological and molecular methods. The predominant species was Enterococcus durans, followed by Enterococcus faecalis and Enterococcus faecium. Over 44% of the isolates were resistant to ciprofloxacin and erythromycin, while 26.2% of the isolates were multi resistant to three or more antibiotics belonging to different families. 185 isolates (29.1%) were susceptible to all 13 of the antibiotics tested. The antibiotic-susceptible isolates were further tested for possible virulence genes and the production of biogenic amines. Finally, five enterococci isolates were found to be antibiotic susceptible with good technological characteristics and without virulence traits or the ability to produce biogenic amines, making them possible candidates for biotechnological application as starter cultures in the dairy industry.",
publisher = "Frontiers Media Sa, Lausanne",
journal = "Frontiers in Microbiology",
title = "Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?",
volume = "6",
doi = "10.3389/fmicb.2015.00954"
}
Terzić-Vidojević, A., Veljović, K., Begović, J., Filipić, B., Popović, D., Tolinacki, M., Miljković, M., Kojić, M.,& Golić, N.. (2015). Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?. in Frontiers in Microbiology
Frontiers Media Sa, Lausanne., 6.
https://doi.org/10.3389/fmicb.2015.00954
Terzić-Vidojević A, Veljović K, Begović J, Filipić B, Popović D, Tolinacki M, Miljković M, Kojić M, Golić N. Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?. in Frontiers in Microbiology. 2015;6.
doi:10.3389/fmicb.2015.00954 .
Terzić-Vidojević, Amarela, Veljović, Katarina, Begović, Jelena, Filipić, Brankica, Popović, Dušanka, Tolinacki, Maja, Miljković, Marija, Kojić, Milan, Golić, Nataša, "Diversity and antibiotic susceptibility of autochthonous dairy enterococci isolates: are they safe candidates for autochthonous starter cultures?" in Frontiers in Microbiology, 6 (2015),
https://doi.org/10.3389/fmicb.2015.00954 . .
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Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese

Uroić, Ksenija; Nikolić, Milica; Kos, Blazenka; Lebos-Pavunc, Andreja; Beganović, Jasna; Lukić, Jovanka; Jovčić, Branko; Filipić, Brankica; Miljković, Marija; Golić, Nataša; Topisirović, Ljubiša; Cadez, Neza; Raspor, Peter; Susković, Jagoda

(Faculty Food Technology Biotechnology, Zagreb, 2014)

TY  - JOUR
AU  - Uroić, Ksenija
AU  - Nikolić, Milica
AU  - Kos, Blazenka
AU  - Lebos-Pavunc, Andreja
AU  - Beganović, Jasna
AU  - Lukić, Jovanka
AU  - Jovčić, Branko
AU  - Filipić, Brankica
AU  - Miljković, Marija
AU  - Golić, Nataša
AU  - Topisirović, Ljubiša
AU  - Cadez, Neza
AU  - Raspor, Peter
AU  - Susković, Jagoda
PY  - 2014
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2197
AB  - The aim of this study is to gain insight into the probiotic potential of autochthonous lactic acid bacteria (LAB) isolated from artisanal fresh soft and white pickled cheeses. Eleven out of 86 LAB isolates from traditionally produced artisanal fresh soft and white pickled cheeses which survived the most rigorous simulated gastrointestinal tract conditions and did not show resistance to antibiotics were subjected to further evaluation for functional probiotic properties. The ability of the examined strains to assimilate cholesterol in the presence of bile salts was strain dependent, with the highest percentage of cholesterol assimilated by strain Lactobacillus brevis BGGO7-28 possessing S-layer proteins on its cell surface. The growth of strains with mannitol or lactulose as the only carbon source was better than with fructooligosaccharides (FOS) and inulin as prebiotic substrates, which should be considered in the production of synbiotics. Moreover, the results demonstrated that the strains were highly adhesive to human enterocyte-like Caco-2 cells and to a lesser extent to HT29-MTX cells, with the exception of strain Lb. brevis BGGO7-28, which showed similar percentage of adhesion to both cell lines. This strain was the only one with the acidic cell surface, while other examined strains have the cell surfaces with electron donor and basic properties. In addition, all selected strains decreased the proliferation of gut-associated lymphoid tissue (GALT) cells, suggesting possible immunomodulatory potential of the isolates. Finally, the number of viable cells in dry active preparations after lyophilisation depended on the lyoprotectant used (inulin, FOS or skimmed milk), as well as on the strain subjected to lyophilisation. In conclusion, the results obtained in this study demonstrate that particular dairy LAB isolates exhibit strain-specific probiotic properties. Thus, they could be further examined as part of mixed autochthonous starter cultures for traditional cheese production under controlled conditions.
PB  - Faculty Food Technology Biotechnology, Zagreb
T2  - Food Technology and Biotechnology
T1  - Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese
VL  - 52
IS  - 2
SP  - 232
EP  - 241
UR  - https://hdl.handle.net/21.15107/rcub_farfar_2197
ER  - 
@article{
author = "Uroić, Ksenija and Nikolić, Milica and Kos, Blazenka and Lebos-Pavunc, Andreja and Beganović, Jasna and Lukić, Jovanka and Jovčić, Branko and Filipić, Brankica and Miljković, Marija and Golić, Nataša and Topisirović, Ljubiša and Cadez, Neza and Raspor, Peter and Susković, Jagoda",
year = "2014",
abstract = "The aim of this study is to gain insight into the probiotic potential of autochthonous lactic acid bacteria (LAB) isolated from artisanal fresh soft and white pickled cheeses. Eleven out of 86 LAB isolates from traditionally produced artisanal fresh soft and white pickled cheeses which survived the most rigorous simulated gastrointestinal tract conditions and did not show resistance to antibiotics were subjected to further evaluation for functional probiotic properties. The ability of the examined strains to assimilate cholesterol in the presence of bile salts was strain dependent, with the highest percentage of cholesterol assimilated by strain Lactobacillus brevis BGGO7-28 possessing S-layer proteins on its cell surface. The growth of strains with mannitol or lactulose as the only carbon source was better than with fructooligosaccharides (FOS) and inulin as prebiotic substrates, which should be considered in the production of synbiotics. Moreover, the results demonstrated that the strains were highly adhesive to human enterocyte-like Caco-2 cells and to a lesser extent to HT29-MTX cells, with the exception of strain Lb. brevis BGGO7-28, which showed similar percentage of adhesion to both cell lines. This strain was the only one with the acidic cell surface, while other examined strains have the cell surfaces with electron donor and basic properties. In addition, all selected strains decreased the proliferation of gut-associated lymphoid tissue (GALT) cells, suggesting possible immunomodulatory potential of the isolates. Finally, the number of viable cells in dry active preparations after lyophilisation depended on the lyoprotectant used (inulin, FOS or skimmed milk), as well as on the strain subjected to lyophilisation. In conclusion, the results obtained in this study demonstrate that particular dairy LAB isolates exhibit strain-specific probiotic properties. Thus, they could be further examined as part of mixed autochthonous starter cultures for traditional cheese production under controlled conditions.",
publisher = "Faculty Food Technology Biotechnology, Zagreb",
journal = "Food Technology and Biotechnology",
title = "Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese",
volume = "52",
number = "2",
pages = "232-241",
url = "https://hdl.handle.net/21.15107/rcub_farfar_2197"
}
Uroić, K., Nikolić, M., Kos, B., Lebos-Pavunc, A., Beganović, J., Lukić, J., Jovčić, B., Filipić, B., Miljković, M., Golić, N., Topisirović, L., Cadez, N., Raspor, P.,& Susković, J.. (2014). Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese. in Food Technology and Biotechnology
Faculty Food Technology Biotechnology, Zagreb., 52(2), 232-241.
https://hdl.handle.net/21.15107/rcub_farfar_2197
Uroić K, Nikolić M, Kos B, Lebos-Pavunc A, Beganović J, Lukić J, Jovčić B, Filipić B, Miljković M, Golić N, Topisirović L, Cadez N, Raspor P, Susković J. Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese. in Food Technology and Biotechnology. 2014;52(2):232-241.
https://hdl.handle.net/21.15107/rcub_farfar_2197 .
Uroić, Ksenija, Nikolić, Milica, Kos, Blazenka, Lebos-Pavunc, Andreja, Beganović, Jasna, Lukić, Jovanka, Jovčić, Branko, Filipić, Brankica, Miljković, Marija, Golić, Nataša, Topisirović, Ljubiša, Cadez, Neza, Raspor, Peter, Susković, Jagoda, "Probiotic Properties of Lactic Acid Bacteria Isolated from Croatian Fresh Soft Cheese and Serbian White Pickled Cheese" in Food Technology and Biotechnology, 52, no. 2 (2014):232-241,
https://hdl.handle.net/21.15107/rcub_farfar_2197 .
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27

Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis

Filipić, Brankica; Jovčić, Branko; Uzelac, Gordana; Miljković, Marija; Antić-Stanković, Jelena; Topisirović, Ljubiša; Golić, Nataša

(Društvo genetičara Srbije, Beograd, 2013)

TY  - JOUR
AU  - Filipić, Brankica
AU  - Jovčić, Branko
AU  - Uzelac, Gordana
AU  - Miljković, Marija
AU  - Antić-Stanković, Jelena
AU  - Topisirović, Ljubiša
AU  - Golić, Nataša
PY  - 2013
UR  - https://farfar.pharmacy.bg.ac.rs/handle/123456789/2045
AB  - The influence of the over-expression of CmbT multidrug resistance transporter on the growth rate of Lactococcus lactis NZ9000 was studied. L. lactis is a lactic acid bacteria (LAB) widely used as a starter culture in dairy industry. Recently characterized CmbT MDR transporter in L. lactis confers resistance to a wide variety of toxic compounds as well as to some clinically relevant antibiotics. In this study, the cmbT gene was over-expressed in the strain L. lactis NZ9000 in the presence of nisin inducer. Over-expression of the cmbT gene in L. lactis NZ9000 was followed by RT-PCR. The obtained results showed that the cmbT gene was successfully over-expressed by addition of sub-inhibitory amounts of nisin. Growth curves of L. lactis NZ9000/pCT50 over-expressing the cmbT gene and L. lactis NZ9000 control strain were followed in the rich medium as well as in the chemically defined medium in the presence solely of methionine (0.084 mM) or mix of methionine and cysteine (8.4 mM and 8.2 mM, respectively). Resulting doubling times revealed that L. lactis NZ9000/pCT50 had higher growth rate comparing to the control strain. This could be a consequence of the CmbT efflux activity, which improves the fitness of the host bacterium through the elimination of toxic compounds from the cell.
AB  - U ovom radu je izučavan uticaj povećane ekspresije cmbT gena, odgovornog za sintezu CmbT MDR transportera, na rast Lactococcus lactis. L. lactis pripada grupi bakterija mlečne kiseline (BMK) i ima veliku primenu u prehrambenoj industriji kao starter kultura. CmbT transporter je nedavno okarakterisan MDR protein soja L. lactis, koji doprinosi rezistenciji na različite toksične agense kao i na neke klinički značajne antibiotike. U ovom radu je cmbT gen višestruko eksprimiran u soju L. lactis NZ9000 dodavanjem nizina kao inducera. Povećana ekspresija cmbT gena je praćena metodom reverzne transkripcije (RT-PCR). Pokazano je da se nakon dodatka subinhibitornih koncentracija nizina u medijum za rast povećava količina sintetisane informacione RNK specifične za cmbT gen. Rast soja L. lactis NZ9000/pCT50, u kome je višestruko eksprimiran cmbT gen i L. lactis NZ9000 kontrolnog soja praćen je u bogatom i hemijski definisanom medijumu u prisustvu samo metionina (0.084 mM) ili kombinacije metionina i cisteina (8.4 mM i 8.2 mM). Praćene su krive rasta oba soja, a nakon izračunavanja odgovarajućih vremena generacije, rezultati su pokazali da L. lactis NZ9000/pCT50, brže raste u odnosu na kontrolni soj. Uočena razlika je najverovatnije posledica aktivnosti CmbT transportera koji doprinosi izbacivanju toksičnih agenasa iz ćelije i na taj način poboljšava adaptivne sposobnosti bakterije koja ga eksprimira i daje joj selektivnu prednost.
PB  - Društvo genetičara Srbije, Beograd
T2  - Genetika, Belgrade
T1  - Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis
T1  - Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis
VL  - 45
IS  - 1
SP  - 197
EP  - 206
DO  - 10.2298/GENSR1301197F
ER  - 
@article{
author = "Filipić, Brankica and Jovčić, Branko and Uzelac, Gordana and Miljković, Marija and Antić-Stanković, Jelena and Topisirović, Ljubiša and Golić, Nataša",
year = "2013",
abstract = "The influence of the over-expression of CmbT multidrug resistance transporter on the growth rate of Lactococcus lactis NZ9000 was studied. L. lactis is a lactic acid bacteria (LAB) widely used as a starter culture in dairy industry. Recently characterized CmbT MDR transporter in L. lactis confers resistance to a wide variety of toxic compounds as well as to some clinically relevant antibiotics. In this study, the cmbT gene was over-expressed in the strain L. lactis NZ9000 in the presence of nisin inducer. Over-expression of the cmbT gene in L. lactis NZ9000 was followed by RT-PCR. The obtained results showed that the cmbT gene was successfully over-expressed by addition of sub-inhibitory amounts of nisin. Growth curves of L. lactis NZ9000/pCT50 over-expressing the cmbT gene and L. lactis NZ9000 control strain were followed in the rich medium as well as in the chemically defined medium in the presence solely of methionine (0.084 mM) or mix of methionine and cysteine (8.4 mM and 8.2 mM, respectively). Resulting doubling times revealed that L. lactis NZ9000/pCT50 had higher growth rate comparing to the control strain. This could be a consequence of the CmbT efflux activity, which improves the fitness of the host bacterium through the elimination of toxic compounds from the cell., U ovom radu je izučavan uticaj povećane ekspresije cmbT gena, odgovornog za sintezu CmbT MDR transportera, na rast Lactococcus lactis. L. lactis pripada grupi bakterija mlečne kiseline (BMK) i ima veliku primenu u prehrambenoj industriji kao starter kultura. CmbT transporter je nedavno okarakterisan MDR protein soja L. lactis, koji doprinosi rezistenciji na različite toksične agense kao i na neke klinički značajne antibiotike. U ovom radu je cmbT gen višestruko eksprimiran u soju L. lactis NZ9000 dodavanjem nizina kao inducera. Povećana ekspresija cmbT gena je praćena metodom reverzne transkripcije (RT-PCR). Pokazano je da se nakon dodatka subinhibitornih koncentracija nizina u medijum za rast povećava količina sintetisane informacione RNK specifične za cmbT gen. Rast soja L. lactis NZ9000/pCT50, u kome je višestruko eksprimiran cmbT gen i L. lactis NZ9000 kontrolnog soja praćen je u bogatom i hemijski definisanom medijumu u prisustvu samo metionina (0.084 mM) ili kombinacije metionina i cisteina (8.4 mM i 8.2 mM). Praćene su krive rasta oba soja, a nakon izračunavanja odgovarajućih vremena generacije, rezultati su pokazali da L. lactis NZ9000/pCT50, brže raste u odnosu na kontrolni soj. Uočena razlika je najverovatnije posledica aktivnosti CmbT transportera koji doprinosi izbacivanju toksičnih agenasa iz ćelije i na taj način poboljšava adaptivne sposobnosti bakterije koja ga eksprimira i daje joj selektivnu prednost.",
publisher = "Društvo genetičara Srbije, Beograd",
journal = "Genetika, Belgrade",
title = "Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis, Uticaj povećane ekspresije CmbT MDR transportera na rast Lactococcus lactis",
volume = "45",
number = "1",
pages = "197-206",
doi = "10.2298/GENSR1301197F"
}
Filipić, B., Jovčić, B., Uzelac, G., Miljković, M., Antić-Stanković, J., Topisirović, L.,& Golić, N.. (2013). Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis. in Genetika, Belgrade
Društvo genetičara Srbije, Beograd., 45(1), 197-206.
https://doi.org/10.2298/GENSR1301197F
Filipić B, Jovčić B, Uzelac G, Miljković M, Antić-Stanković J, Topisirović L, Golić N. Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis. in Genetika, Belgrade. 2013;45(1):197-206.
doi:10.2298/GENSR1301197F .
Filipić, Brankica, Jovčić, Branko, Uzelac, Gordana, Miljković, Marija, Antić-Stanković, Jelena, Topisirović, Ljubiša, Golić, Nataša, "Over-expressed CmbT multidrug resistance transporter improves the fitness of Lactococcus lactis" in Genetika, Belgrade, 45, no. 1 (2013):197-206,
https://doi.org/10.2298/GENSR1301197F . .
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