Abstract
Lactobacillus helveticus MTCC 5463 was isolated from a vaginal swab from a healthy adult female. The strain exhibited potential probiotic properties, with their beneficial role in the gastrointestinal tract and their ability to reduce cholesterol and stimulate immunity. We sequenced the whole genome and compared it with the published genome sequence of Lactobacillus helveticus DPC4571.
GENOME ANNOUNCEMENT
Lactobacillus helveticus is present in fermented foods and is also used as a probiotic (4, 10, 12). Researchers have evaluated the effect of L. helveticus against diseases such as cancer and intestinal inflammation. Milk fermented with L. helveticus R389 delayed breast tumor growth by decreasing interleukin-6 (IL-6) and increasing IL-10 in serum, mammary glands, and tumor-infiltrating immune cells (5). Lactobacillus helveticus strains are normally isolated from milk products and intestinal microflora (6, 14). The L. helveticus MTCC 5463 strain was originally isolated from the vaginal tract of a healthy adult female in India at Anand Agricultural University (9). The L. helveticus MTCC 5463 strain, earlier known as Lactobacillus acidophilus V3 (based on biochemical characteristics) was able to grow in the presence of 0.3% sodium taurocholate, deconjugate bile acids, and reduce cholesterol in vitro (1). The strain exhibited significant antimicrobial activity against Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella enterica serovar Typhi, and Escherichia coli (7). The strain produced extracellular polysaccharide and was able to adhere to cells of the human carcinoma cell line HT29. A hypocholesterolemic effect of L. helveticus MTCC 5463 was reported in human subjects with different cholesterol levels (2). The strain has also shown positive immunomodulating effects in a chick model (13).
The whole-genome sequencing of L. helveticus MTCC 5463 was performed using GS-FLX Titanium reagents (11). The data generated from the genomic library contained 119,569 reads, and assembly generated a 1,911,350-bp single chromosome. The genome annotation and comparative analysis of the genome were done with the published genome of strain DPC4571 (3). The genomic sequence of L. helveticus MTCC 5463 was somewhat smaller than those of L. helveticus DPC4571 (2.08 Mb) and L. helveticus H10 (2.14 Mb). In total, 2,046 coding sequence (CDS) regions and 71 RNA genes were reported. Of the 71 RNA genes, 59 coded for tRNA, 8 for rRNA, and 4 for 5S RNA. The CDS regions were slightly fewer than in L. helveticus H10 (2,049) and L. helveticus DPC4571 (2,238). Metabolic reconstruction subsystems were assembled to create a metabolic reaction network for L. helveticus MTCC 5463 and L. helveticus DPC4571.
The subsystem analysis revealed a common subsystem structure between L. helveticus MTCC 5463 and L. helveticus DPC4571 for seven subsystems, viz., photosynthesis, iron acquisition and metabolism, motility and chemotaxis, secondary metabolism, stress response, nitrogen metabolism, and dormancy and sporulation. The considerable variation observed in the remaining subsystems indicated biochemical heterogeneity and capabilities of the strain in substrate utilization and processing. The L. helveticus MTCC 5463 genome has 57 genes which are absent in the L. helveticus DPC4571 genome, comprising 15 major categories; 12 of these are for carbohydrate utilization. The results are indicative of a diverse carbohydrate utilization pattern for L. helveticus MTCC 5463. The presence of biotin synthesis genes and differences in cofactors, vitamins, prosthetic groups, and pigments suggest the differential ability of the strain in the production of such bioactive compounds, in contrast to L. helveticus DPC4571.
Nucleotide sequence accession number.
The complete sequence of the L. helveticus MTCC 5463 genome can be accessed under the GenBank accession numbers AEYL01000001 to AEYL01000593.
Acknowledgments
The project was funded by the Indian Council of Agricultural Research under the Niche Area of Excellence program.
Footnotes
Published ahead of print on 24 June 2011.
REFERENCES
- 1. Ashar M. N., Prajapati J. B. 1998. Bile tolerance, bile deconjugation and cholesterol reducing properties of dietary lactobacilli. Indian J. Microbiol. 38:145–148 [Google Scholar]
- 2. Ashar M. N., Prajapati J. B. 2000. Verification of hypocholesterolemic effect of fermented milk on human subjects with different cholesterol level. Folia Microbiol. 45:263–268 [DOI] [PubMed] [Google Scholar]
- 3. Callanan M., et al. 2008. Genome sequence of Lactobacillus helveticus, an organism distinguished by selective gene loss and insertion sequence element expansion. J. Bacteriol. 190:727–735 [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4. Champagne C. P., Tompkins T. A., Buckley N. D., Green-Johnson J. M. 2010. Effect of fermentation by pure and mixed cultures of Streptococcus thermophiles and Lactobacillus helveticus on isoflavone and B-vitamin content of a fermented soy beverage. Food Microbiol. 27:968–972 [DOI] [PubMed] [Google Scholar]
- 5. de Moreno de Leblanc A., Perdigón G. 2010. The application of probiotic fermented milks in cancer and intestinal inflammation. Proc. Nutr. Soc. 69:421–428 [DOI] [PubMed] [Google Scholar]
- 6. Frece J., et al. 2009. Synbiotic effect of Lactobacillus helveticus M92 and prebiotics on the intestinal microflora and immune system of mice. J. Dairy Res. 76:98–104 [DOI] [PubMed] [Google Scholar]
- 7. Khedekar C. D., Dave J. M., Sannabhadti S. S. 1990. Antibacterial activity of human strains of Lactobacillus acidophilus grown in milk against selected pathogenic and spoilage type bacteria. Cultured Dairy Prod. J. 25:29–31 [Google Scholar]
- 8. Reference deleted.
- 9. Khedkar C. D., Dave J. M., Sannabhadti S. S. 1991. Incidence of Lactobacillus acidophilus in human vaginal and gastrointestinal tracts. Indian J. Comp. Microbiol. Immunol. Infect. Dis. 12:107–109 [Google Scholar]
- 10. Kilpi E. R., Kahala M. M., Steele J. L., Pihlanto A. M., Joutsjoki V. V. 2007. Angiotensin I-converting enzyme inhibitory activity in milk fermented by wild-type and peptidase-deletion derivatives of Lactobacillus helveticus CNRZ32. Int. Dairy J. 17:976–984 [Google Scholar]
- 11. Margulies M., et al. 2005. Genome sequencing in microfabricated high density picolitre reactors. Nature 437:376–380 [DOI] [PMC free article] [PubMed] [Google Scholar]
- 12. Messaoudi M., et al. 2011. Assessment of psychotropic-like properties of a probiotic formulation (Lactobacillus helveticus R0052 and Bifidobacterium longum R0175) in rats and human subjects. Br. J. Nutr. 105:755–764 [DOI] [PubMed] [Google Scholar]
- 13. Patidar S. K., Prajapati J. B. 1999. Effect of feeding lactobacilli on serum antibody titre and faecal flora in chicks. Microbiol. Alim. Nutr. 17:145–154 [Google Scholar]
- 14. Zhao W., et al. 2011. Complete genome sequence of Lactobacillus helveticus H10. J. Bacteriol. 193:2666–2667 [DOI] [PMC free article] [PubMed] [Google Scholar]