Sie können Operatoren mit Ihrer Suchanfrage kombinieren, um diese noch präziser einzugrenzen. Klicken Sie auf den Suchoperator, um eine Erklärung seiner Funktionsweise anzuzeigen.
Findet Dokumente, in denen beide Begriffe in beliebiger Reihenfolge innerhalb von maximal n Worten zueinander stehen. Empfehlung: Wählen Sie zwischen 15 und 30 als maximale Wortanzahl (z.B. NEAR(hybrid, antrieb, 20)).
Findet Dokumente, in denen der Begriff in Wortvarianten vorkommt, wobei diese VOR, HINTER oder VOR und HINTER dem Suchbegriff anschließen können (z.B., leichtbau*, *leichtbau, *leichtbau*).
Complete genome sequence of Lactobacillus pentosus SLC13, isolated from mustard pickles, a potential probiotic strain with antimicrobial activity against foodborne pathogenic microorganisms
Lactobacillus pentosus SLC13 is a high exopolysaccharide (EPS)-producing strain with broad-spectrum antimicrobial activity and the ability to grow in simulated gastrointestinal conditions. SLC13 was isolated from mustard pickles in Taiwan for potential probiotic applications. To better understand the molecular base for its antimicrobial activity and high EPS production, entire genome of SLC13 was determined by PacBio SMRT sequencing.
Results
L. pentosus SLC13 contains a genome with a 3,520,510-bp chromosome and a 62,498-bp plasmid. GC content of the complete genome was 46.5% and that of plasmid pSLC13 was 41.3%. Sequences were annotated at the RAST prokaryotic genome annotation server, and the results showed that the genome contained 3172 coding sequences and 82 RNA genes. Seventy-six protein-coding sequences were identified on the plasmid pSLC13. A plantaricin gene cluster, which is responsible for bacteriosins biosynthesis and could be associated with its broad-spectrum antimicrobial activity, was identified based on comparative genomic analysis. Two gene clusters involved in EPS production were also identified.
Conclusion
This genomic sequence might contribute to a future application of this strain as probiotic in productive livestock potentially inhibiting competing and pathogenic organisms.
The beneficial effects of lactic acid bacteria (LAB) and its derivative exopolysaccharide (EPS), include the prevention and treatment of diarrheal disease, prevention of infections, antitumor activity, immunomodulation, prevention and treatment of allergies, and alleviation of lactose intolerance [1‐6]. In our previous study, we isolated a high EPS-producing Lactobacillus pentosus strain, SLC13, from mustard pickles in Taiwan for potential probiotic applications [7]. SLC13 showed high resistance to bile salts, pH 3.0 PBS, and simulated gastrointestinal conditions. Moreover, the results of antimicrobial activity tests revealed that SLC13 showed high inhibitory activity against the growth of clinical important pathogens, including Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis, Listeria monocytogenes Scott A, Yersinia enterocolitica, Escherichia coli O157:H7, and Salmonella enterica sv. Typhimurium [7]. Here, the complete genome sequence of SLC13 was characterized to identify potential gene(s), which is (are) responsible for high EPS production or could be associated with its broad-spectrum antimicrobial activity.
Methods
Whole genome sequencing, assembly and annotation
The genomic DNA of SLC13 was extracted using the DNeasy Blood and Tissue Kit (QIAGEN, Germany), according to the manufacturer’s instructions. Total DNA was subjected to quality control by 2% agarose gel electrophoresis and quantified by a NanoDrop™ spectrophotometer. L. pentosus SLC13 was sequenced using the PacBio RS II platform (Pacific Biosciences, USA), and the reads were assembled using HGAP version 3.0. Sequences were further annotated at the RAST prokaryotic genome annotation server (http://rast.nmpdr.org/) [8].
Anzeige
Analysis of antibiotic resistance genes, bacteriocin synthesis genes, and EPS-gene cluster
ResFinder databases (http://www.genomicepidemiology.org/) were used to find the antibiotic resistance genes present in the plasmid pSLC13. The first step of the bacteriocin identification workflow was created by merging the BACTIBASE databases using BLASTN [9] and BAGEL (class I, II and III) [10]. RAST server was used for identifying eps gene cluster by comparative genomic analysis.
Quality assurance
The genomic DNA used for sequencing was isolated from a single colony of the SLC13. The 16S rRNA gene was sequenced and BLAST was performed against the NCBI database. In addition, the raw read sequences were selected and assembled only when they satisfied the following criteria: minimum subread length, 500; minimum polymerase read quality, 0.8; and minimum polymerase read length, 100.
Results and discussion
General features
L. pentosus SLC13 was sequenced using the PacBio RS II platform, generating a library containing 58,744 single reads with an average length of 3936 bp. Reads were assembled and returned two contigs with the head segment was almost identical to the tail segment, indicating the circular nature of the contigs. As shown in Fig. 1 and Table 1, the complete genome sequence of SLC13 was composed of a circular 3,520,510-bp chromosome and one 62,498-bp plasmid named as pSLC13. GC content of the complete genome was 46.5% and that of plasmid pSLC13 was 41.3%. The GC content and size of SLC13 chromosome was similar to other Lactobacillus strains, including L. plantarum GB-LP1 (3,040,388 bp, GC content: 44.9%) [11], L. rhamnosus CNCM I-3698 (2,966,480 bp, GC content: 46.69%) [12], L. casei BD-II (3,069,926 bp, GC content: 46.34%) [13], L. pentosus KCAI (3,418,159 bp, GC content: 46.4%) [14], but not L. jojnsonii FI9785 (1,755,993 bp, GC content: 34.49%) [15] (Table 1).
Fig. 1
Circular genome map of L. pentosus SLC13. a Chromosome. b pSLC13. The scales indicate the location in Mbp, starting with the initial coding region. From the innermost circles, circle (1) GC content, plotted using a sliding window. Circle (2) shows the GC skew (G-C/G+C). The value is plotted as the deviation from the average GC skew of the entire sequence. Circle (3, 4) illustrate the coding sequences, 3 is backward strand, 4 is forward strand
Comparison of the features of Lactobacillus spp. genome and probiotic potential
Attributes
Lactobacillus strains
Species
L. pentosus
L. plantarum
L. rhamnosus
L. casei
L. pentosus
L. johnsonii
Strain
SLC13
GB-LP1
CNCM I-3698
BD-II
KCA1
FI9785
Size (bp)
3,520,510
3,040,388
2,966,480
3,069,926
3,418,159
1,755,993
GC content (%)
46.5
44.9
46.69
46.34
46.4
34.49
Plasmid (no.)
1 (pSLC13: 62,498 bp)
–
–
1 (57,362 bp)
–
2 (p9785S: 3471 bp; p9785L: 25,652 bp)
RNAs (no.)
82
79
55
64
80
57
Coding sequences
3172 (76 in plasmids)
2899
2939
3139 (65 in plasmids)
2967
1710 (3 in p9785S; 25 in p9785L)
Resource (country)
Mustard pickles (Taiwan)
Traditional fermented food (Korea)
Goat rumen (UK)
Homemade koumiss (China)
Human vagina (Netherlands)
Poultry-derived (UK)
Probiotic potentials
Immune response, antibacterial activity
Immune response, suppression of pathogen growth, and antitoxin effects
Antagonistic activities against zoonotic pathogens
Reduce the blood lipid level and regulate the cardiovascular system
Produce biosurfactants, hydrogen peroxide (H2O2), and inhibit the growth of intestinal and urogenital pathogens
Immunomodulation, and competitive exclusion of pathogens
RAST annotation results showed that the genome contained 3172 coding sequences and 82 RNA genes (Table 1, Fig. 2). Moreover, all protein coding sequences were functionally annotated by RAST server in 350 subsystems (Fig. 2). Seventy-six protein-coding sequences were identified on the plasmids pSLC13. The sequence blast results revealed that pSLC13 showed high similarity to plasmid pLB1106-3 in L. brevis strain SRCM101106 (accession number: CP021675). In our previous study, we showed that SLC13 was resistant to penicillin G, cephalothin, cloxacillin, novobiocin, vancomycin, polymyxin B, rifampicin, tetercycline, kanamycin, gentamycin, neomycin, and streptomycin [7]. ResFinder databases were used to find the antibiotic resistance genes present in the plasmid pSLC13, and the results showed no antibiotic resistance genes were identified in pSLC13. Therefore, there is no further consideration for transmission of antibiotic resistant determinants by SLC13 as a candidate for probiotic development.
Fig. 2
Subsystem distribution of L. pentosus SLC13 based on RAST annotation server. Out of 3172 coding sequences predicted by RAST server, the subsystem coverage is 44% which contributes to a total of 350 subsystems. The green bar of the subsystem coverage indicates the percentage of the proteins included in the subsystems while the blue bar refers to the percentage of the proteins that are not included in the subsystems
SLC 13 isolated from mustard pickles produced the largest amount of EPS (0.43 ± 0.04 g/l) among the 39 collected LAB isolates in our previous report [7]. To explore the gene cluster(s) that is (are) responsible for this phenotype, the comparative genomic analysis was performed by RAST server, and the genes that could be related to high-EPS biosynthesis were annotated in detail (Additional file 1: Figure S1). The genome of the SLC13 encoded 2 recognizable exopolysaccharide (eps) gene clusters. Comparison of the genome of L. pentosus SLC13 with that of strain L. plantarum WCFS1, indicated that the two EPS synthesis clusters assigned in SLC13 is highly conserved to the L. plantarum WCFS1 (Additional file 1: Figure S1). However, whether these two eps gene clusters responsible for high-EPS production in SLC13 is still unknown. Currently, Lee et al. reported that the abundance and sugar compositions of EPS diverse in three L. plantarum strains [16]. The diverse of sugar composition may affect its impact on probiotic-host interaction, for example, host cell adhesion [16]. Therefore, the composition of SLC13-EPS remains to be characterized in the future.
Plantaricin (pln) gene cluster identification
A plantaricin (pln) gene cluster for bacteriocin synthesis in SLC13 identified by RAST server was shown in Additional file 1: Figure S2a. Further investigation was then conducted with BAGEL3, a web-based bacteriocin mining tool. The result from BAGEL3 showed a putative class II bacteriocin, Pediocin PA-1 immunity protein, was identified on SLC13 chromosome, as shown in Additional file 1: Figure S2b (gene start position, 2,037,423 on the chromosome; length, 112 amino acids). Pediocin PA-1 was previous identified in probiotic Pediococcus acidilactici with antimicrobial activity against Listeria spp., Oenococcus oeni and other wine bacteria [17, 18]. However, the antimicrobial activity of pediocin in SLC13 remains to be verified.
Identification of genes coding for virulence factors
RAST annotation results showed that four virulence factors were found in the SLC13 chromosome, including heat shock protein 33, Jag, YidD, and YidC. Heat shock protein 33 is an adhesin important for adhesion and colonization of the Streptococcus pyogenes on epithelial cells [19]. Jag, YidD, and YidC are part of the Mycobacterium virulence operon with unclear function. Currently, Thakur et al. reported that the translocase YidC controls respiratory metabolism in M. tuberculosis and is required for the intracellular survival of M. tuberculosis in human macrophages [20]. A type II toxin-antitoxin system, mRNA interferase toxin, which is ubiquitous in Lactobacillus strains with the ability to inhibit E. coli growth, was found in pSLC13 [21]. However, the function of virulence factors identified in SLC13 remains to be determined.
Together, the complete genome sequence of SLC13 allows us to better understand molecular basis of its antimicrobial activity, high EPS production and probiotic potentials. Future studies are needed to verify the probiotic properties and safety of this strain for its industry application.
Authors' contributions
MLH, JYH and CYK wrote the manuscript. Automatic annotation and genome submission preparation were executed by MLH, JYH, and CYK. All authors read and approved the final manuscript.
Acknowledgements
Not applicable.
Competing of interests
The authors declare that they have no competing interests.
Availability of data and materials
The completed genome sequence of L. pentosus SLC13 has been deposited in the GenBank database under Accession Numbers CP022130 (chromosome) and CP022131 (plasmid).
Anzeige
Consent for publication
Not applicable.
Ethics approval and consent to participate
Not applicable.
Funding
This study was supported by Grants NSC95-2313-B005-054 and MOST 106-2917-I-564-070 from the Ministry of Science and Technology, R.O.C.
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Anzeige
Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Complete genome sequence of Lactobacillus pentosus SLC13, isolated from mustard pickles, a potential probiotic strain with antimicrobial activity against foodborne pathogenic microorganisms
Verfasst von
Min-Lang Huang
Jing-Yao Huang
Cheng-Yen Kao
Tony J. Fang
Stiles ME, Holzapfel WH. Lactic acid bacteria of foods and their current taxonomy. Int J Food Microbiol. 1997;36:1–29.CrossRefPubMed
2.
Saarela M, Mogensen G, Fonden R, Matto J, Mattila-Sandholm T. Probiotic bacteria: safety, functional and technological properties. J Biotechnol. 2000;84:197–215.CrossRefPubMed
3.
Ruiz-Bravo A, Jimenez-Valera M, Moreno E, Guerra V, Ramos-Cormenzana A. Biological response modifier activity of an exopolysaccharide from Paenibacillus jamilae CP-7. Clin Diagn Lab Immunol. 2001;8:706–10.PubMedPubMedCentral
4.
Gill HS, Guarner F. Probiotics and human health: a clinical perspective. Postgrad Med J. 2004;80:516–26.CrossRefPubMedPubMedCentral
5.
Maldonado NC, de Ruiz CS, Otero MC, Sesma F, Nader-Macias ME. Lactic acid bacteria isolated from young calves-characterization and potential as probiotics. Res Vet Sci. 2012;92:342–9.CrossRefPubMed
6.
Wang K, Li W, Rui X, Chen X, Jiang M, Dong M. Characterization of a novel exopolysaccharide with antitumor activity from Lactobacillus plantarum 70810. Int J Biol Macromol. 2014;63:133–9.CrossRefPubMed
7.
Huang JY, Kao CY, Liu WS, Fang TJ. Characterization of high exopolysaccharide-producing Lactobacillus strains isolated from mustard pickles for potential probiotic applications. Int Microbiol. 2017;20:75–84.PubMed
8.
Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O. The RAST server: rapid annotations using subsystems technology. BMC Genom. 2008;9:75.CrossRef
9.
Hammami R, Zouhir A, Le Lay C, Ben Hamida J, Fliss I. BACTIBASE second release: a database and tool platform for bacteriocin characterization. BMC Microbiol. 2010;10:22.CrossRefPubMedPubMedCentral
10.
de Jong A, van Hijum SA, Bijlsma JJ, Kok J, Kuipers OP. BAGEL: a web-based bacteriocin genome mining tool. Nucleic Acids Res. 2006;34:W273–9.CrossRefPubMedPubMedCentral
11.
Yu J, Ahn S, Kim K, Caetano-Anolles K, Lee C, Kang J, Cho K, Yoon SH, Kang DK, Kim H. Comparative Genomic Analysis of Lactobacillus plantarum GB-LP1 Isolated from Traditional Korean Fermented Food. J Microbiol Biotechnol. 2017;27:1419–27.PubMed
12.
Tareb R, Bernardeau M, Vernoux JP. Genome sequence of Lactobacillus rhamnosus strain CNCM I-3698. Genome Announc. 2015;3:e00582–15.CrossRefPubMedPubMedCentral
13.
Ai L, Chen C, Zhou F, Wang L, Zhang H, Chen W, Guo B. Complete genome sequence of the probiotic strain Lactobacillus casei BD-II. J Bacteriol. 2011;193:3160–1.CrossRefPubMedPubMedCentral
14.
Anukam KC, Macklaim JM, Gloor GB, Reid G, Boekhorst J, Renckens B, van Hijum SA, Siezen RJ. Genome sequence of Lactobacillus pentosus KCA1: vaginal isolate from a healthy premenopausal woman. PLoS ONE. 2013;8:e59239.CrossRefPubMedPubMedCentral
15.
Wegmann U, Overweg K, Horn N, Goesmann A, Narbad A, Gasson MJ, Shearman C. Complete genome sequence of Lactobacillus johnsonii FI9785, a competitive exclusion agent against pathogens in poultry. J Bacteriol. 2009;191:7142–3.CrossRefPubMedPubMedCentral
16.
Lee IC, Caggianiello G, van Swam II, Taverne N, Meijerink M, Bron PA, Spano G, Kleerebezem M. Strain-specific features of extracellular polysaccharides and their impact on Lactobacillus plantarum-host interactions. Appl Environ Microbiol. 2016;82:3959–70.CrossRefPubMedPubMedCentral
17.
Díez L, Rojo-Bezares B, Zarazaga M, Rodríguez JM, Torres C, Ruiz-Larrea F. Antimicrobial activity of pediocin PA-1 against Oenococcus oeni and other wine bacteria. Food Microbiol. 2012;31:167–72.CrossRefPubMed
18.
Le Blay G, Hammami R, Lacroix C, Fliss I. Stability and inhibitory activity of pediocin PA-1 against Listeria sp. in simulated physiological conditions of the human terminal ileum. Probiotics Antimicrob Proteins. 2012;4:250–8.CrossRefPubMed
19.
Murakami J, Terao Y, Morisaki I, Hamada S, Kawabata S. Group A streptococcus adheres to pharyngeal epithelial cells with salivary proline-rich proteins via GrpE chaperone protein. J Biol Chem. 2012;287:22266–75.CrossRefPubMedPubMedCentral
20.
Thakur P, Gantasala NP, Choudhary E, Singh N, Abdin MZ, Agarwal N. The preprotein translocase YidC controls respiratory metabolism in Mycobacterium tuberculosis. Sci Rep. 2016;6:24998.CrossRefPubMedPubMedCentral
21.
Klimina KM, Kjasova DK, Poluektova EU, Krugel H, Leuschner Y, Saluz HP, Danilenko VN. Identification and characterization of toxin-antitoxin systems in strains of Lactobacillus rhamnosus isolated from humans. Anaerobe. 2013;22:82–9.CrossRefPubMed
Von Cortison über Biologika zu CAR‑T‑Zellen: Was ändert sich aktuell in der Rheumatologie? Prof. Elisabeth Märker-Hermann wirft gemeinsam mit Moderator Prof. Markus M. Lerch einen Blick auf die Behandlung von Polymyalgia rheumatica, rheumatoider Arthritis oder Lupus erythematodes. Einige Paradigmenwechsel sind hier im Gange. Die Rheumatologin betont außerdem: Einfache Fragen und wenige Basisuntersuchungen geben erste wichtige diagnostische Hinweise.
Bei einem nach Radiotherapie lokal rezidivierten Prostatakarzinom sind fokale Salvage-Therapien mit einer guten Prognose verbunden: Das krebsspezifische Zehn-Jahres-Überleben ist einem retrospektiven Vergleich zufolge ebenso hoch wie nach Salvage-Prostatektomie.
Durch Hinzunahme des Glukokortikoid-Rezeptor-Antagonisten Relacorilant zu nab-Paclitaxel wird bei Frauen mit platinresistentem Ovarialkarzinom nicht nur das progressionsfreie, sondern auch das Gesamtüberleben verlängert. Laut finaler Analyse der ROSELLA-Studie gewinnen sie vier Monate an Lebenszeit.
Immunvermittelte Hautreaktionen gehören zu den häufigsten Nebenwirkungen von Immun‑Checkpoint‑Inhibitoren. Eine offene Phase‑2‑Studie untersuchte den JAK‑1‑Inhibitor Upadacitinib bei schwerer ICI‑assoziierter Dermatitis. Die Hautsymptome gingen rasch zurück, schwerwiegende therapieassoziierte Nebenwirkungen wurden nicht beobachtet.
