Here, we report the ndraft genome sequences of Lactobacillus delbrueckii subsp. bulgaricus strains CBC-LB69 and CBC-LB8. The strains were isolated from naturally processed, homemade dairy foods in Bulgaria. The two genome assemblies each resulted in 39 contigs, with total lengths of 1,752,493 and 1,759,908 bp and GC contents of 49.80% and 49.90%, respectively.
ABSTRACT
Here, we report the draft genome sequences of Lactobacillus delbrueckii subsp. bulgaricus strains CBC-LB69 and CBC-LB8. The strains were isolated from naturally processed, homemade dairy foods in Bulgaria. The two genome assemblies each resulted in 39 contigs with total lengths of 1,752,493 and 1,759,908 bp and GC contents of 49.80% and 49.90%, respectively.
ANNOUNCEMENT
Lactobacillus delbrueckii subsp. bulgaricus is a common lactic acid bacterium widely used as a starter culture in yogurt and dairy products (1, 2) and was reported to have health benefits such as anti-Helicobacter pylori activity (3), immunity enhancement (4–6), and reduction of allergies to dairy products (7).
Strains CBC-LB69 and CBC-LB8 were isolated from naturally processed, homemade dairy foods from ecologically clean preserves in industrial pollution-free and chemically untampered areas. Strain CBC-LB69 was isolated from Pirin Mountain, Bulgaria, in September 1969, and strain CBC-LB8 was isolated from Strandja Mountain, Bulgaria, in April 1980. Both strains were isolated on MRS agar plates incubated anaerobically at 44°C for 72 h (8) and were found to survive and colonize the human intestinal tract (9).
In this paper, we present the genome sequences of Lactobacillus delbrueckii subsp. bulgaricus strains CBC-LB69 and CBC-LB8. The genomes were sequenced to better understand the genetic basis of their probiotic health effects and their safety for human consumption.
Both strains were obtained from Chandler Biopharmaceutical Corporation as lyophilized powder. Genomic DNA was extracted from 50 mg of lyophilized powder using a NucleoSpin food kit (740945.50; Macherey-Nagel, Germany). The DNA was quantified using a Qubit 4.0 fluorometer and submitted to the Advanced Analysis Centre, University of Guelph (Guelph, ON, Canada), for library preparation and sequencing on the Illumina MiSeq platform using an Illumina Nextera XT kit and Illumina MiSeq v3 600-cycle reagent kit (2 × 300-bp reads).
The sequencing data were analyzed using CLC Genomics Workbench v20.0.3 (Qiagen Bioinformatics). Default parameters were used except where otherwise noted. The reads were quality trimmed to remove the low-quality sequences (limit = 0.05 base-calling error probability), allowing a maximum of 2 ambiguous nucleotides. The total number of paired-end reads from strains CBC-LB69 and CBC-LB8 were 2,308,638 and 2,134,850 before quality trimming and 2,279,307 and 2,122,703 after quality trimming, respectively. High-quality reads with totals of 270,907,719 and 322,302,876 bases were assembled using CLC de novo assembly with default parameters, achieving >150× coverage. Each genome assembly resulted in 39 contigs. The CBC-LB69 draft genome sequence has a total length of 1,752,493 bp, a GC content of 49.80%, and an N 50 value of 93,636 bp. The CBC-LB8 draft genome sequence has a total length of 1,759,908 bp, a GC content of 49.90%, and an N 50 value of 87,463 bp.
Full-length 16S rRNA gene sequences were extracted from the genome sequences using the ContEst16S tool (10) and were BLAST searched (using MegaBLAST against the nucleotide collection database and the rRNA/ITS databases) on GenBank to confirm the species identity as Lactobacillus delbrueckii subsp. bulgaricus (11). The genomes were annotated using the NCBI Prokaryotic Genome Annotation Pipeline (PGAP) (12) v4.11 (http://www.ncbi.nlm.nih.gov/genome/annotation_prok/). CBC-LB69 was found to contain a total of 1,591 coding genes and 60 tRNA genes, while CBC-LB8 contained 1,586 coding genes and 61 tRNA genes.
Data availability.
These whole-genome shotgun projects have been deposited at DDBJ/ENA/GenBank under the accession numbers JABWGR000000000 and JABWOS000000000. The versions described in this paper are the first versions, JABWGR010000000 and JABWOS010000000. The raw files were deposited in the SRA under the accession numbers SRR12037316 and SRR12037315.
ACKNOWLEDGMENT
The Natural Health Product Research Alliance (NHPRA), University of Guelph, supported this study.
REFERENCES
- 1. Torriani S, Vescovo M, Dicks LMT. 1997. Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus: a review. Ann Microbiol Enzimol 47:29–52. [Google Scholar]
- 2. Dan T, Ren W, Liu Y, Tian J, Chen H, Li T, Liu W. 2019. Volatile flavor compounds profile and fermentation characteristics of milk fermented by Lactobacillus delbrueckii subsp. bulgaricus . Front Microbiol 10:2183. doi: 10.3389/fmicb.2019.02183. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3. Boyanova L, Gergova G, Markovska R, Yordanov D, Mitov I. 2017. Bacteriocin-like inhibitory activities of seven Lactobacillus delbrueckii subsp bulgaricus strains against antibiotic susceptible and resistant Helicobacter pylori strains. Lett Appl Microbiol 65:469–474. doi: 10.1111/lam.12807. [DOI] [PubMed] [Google Scholar]
- 4. Moro-Garcia MA, Alonso-Arias R, Baltadjieva M, Benitez CF, Barrial MAF, Ruisanchez ED, Santos RA, Sanchez MA, Mijan JS, Lopez-Larrea C. 2013. Oral supplementation with Lactobacillus delbrueckii subsp bulgaricus 8481 enhances systemic immunity in elderly subjects. Age (Dordr) 35:1311–1326. doi: 10.1007/s11357-012-9434-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 5. Makino S, Ikegami S, Kume A, Horiuchi H, Sasaki H, Orii N. 2010. Reducing the risk of infection in the elderly by dietary intake of yoghurt fermented with Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1. Br J Nutr 104:998–1006. doi: 10.1017/S000711451000173X. [DOI] [PubMed] [Google Scholar]
- 6. Castanheira LG, Castro JMDA, Martins-Filho ODA, Nicoli JR, Vieira LQ, Afonso LCC. 2007. Lactobacillus delbrueckii as a potential skin adjuvant for induction of type 1 immune responses. Front Biosci 12:1300–1307. doi: 10.2741/2148. [DOI] [PubMed] [Google Scholar]
- 7. Pescuma M, Hébert EM, Rabesona H, Drouet M, Choiset Y, Haertlé T, Mozzi F, de Valdez GF, Chobert J-M. 2011. Proteolytic action of Lactobacillus delbrueckii subsp. bulgaricus CRL 656 reduces antigenic response to bovine β-lactoglobulin. Food Chem 127:487–492. doi: 10.1016/j.foodchem.2011.01.029. [DOI] [PubMed] [Google Scholar]
- 8. Michaylova M, Minkova S, Kimura K, Sasaki T, Isawa K. 2007. Isolation and characterization of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus from plants in Bulgaria. FEMS Microbiol Lett 269:160–169. doi: 10.1111/j.1574-6968.2007.00631.x. [DOI] [PubMed] [Google Scholar]
- 9. Vázquez C, Botella-Carretero JI, García-Albiach R, Pozuelo MJ, Rodríguez-Baños M, Baquero F, Baltadjieva MA, del Campo R. 2013. Screening in a Lactobacillus delbrueckii subsp. bulgaricus collection to select a strain able to survive to the human intestinal tract. Nutr Hosp 28:1227–1235. doi: 10.3305/nh.2013.28.4.6540. [DOI] [PubMed] [Google Scholar]
- 10. Lee I, Chalita M, Ha S-M, Na S-I, Yoon S-H, Chun J. 2017. ContEst16S: an algorithm that identifies contaminated prokaryotic genomes using 16S RNA gene sequences. Int J Syst Evol Microbiol 67:2053–2057. doi: 10.1099/ijsem.0.001872. [DOI] [PubMed] [Google Scholar]
- 11. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. 1990. Basic local alignment search tool. J Mol Biol 215:403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
- 12. Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP, Zaslavsky L, Lomsadze A, Pruitt KD, Borodovsky M, Ostell J. 2016. NCBI Prokaryotic Genome Annotation Pipeline. Nucleic Acids Res 44:6614–6624. doi: 10.1093/nar/gkw569. [DOI] [PMC free article] [PubMed] [Google Scholar]
Associated Data
This section collects any data citations, data availability statements, or supplementary materials included in this article.
Data Availability Statement
These whole-genome shotgun projects have been deposited at DDBJ/ENA/GenBank under the accession numbers JABWGR000000000 and JABWOS000000000. The versions described in this paper are the first versions, JABWGR010000000 and JABWOS010000000. The raw files were deposited in the SRA under the accession numbers SRR12037316 and SRR12037315.
