Antonie van Leeuwenhoek

, Volume 111, Issue 7, pp 1149–1156 | Cite as

Lactobacillus kosoi sp. nov., a fructophilic species isolated from kôso, a Japanese sugar-vegetable fermented beverage

  • Tai-Ying Chiou
  • Wataru Suda
  • Kenshiro Oshima
  • Masahira Hattori
  • Chiaki Matsuzaki
  • Kenji Yamamoto
  • Tomoya Takahashi
Original Paper


A novel Gram-positive, fructophilic, catalase negative, and rod-shaped strain, designated strain 10HT was isolated from kôso, a Japanese sugar-vegetable fermented beverage obtained from a food processing factory in Saku City, Nagano Prefecture, Japan. Phylogenetic analysis based on 16S rRNA gene sequences revealed strain 10HT to belong to the genus Lactobacillus, with closely related type strains being Lactobacillus kunkeei YH-15T (95.5% sequence similarity), Lactobacillus ozensis Mizu2-1T (95.4% sequence similarity), and Lactobacillus apinorum Fhon13NT (95.3% sequence similarity). The isolate was found to grow at 18–39 °C (optimum 27 °C), pH 4.0–7.0 (optimum pH 6.5) and in the presence of 0–2% NaCl (optimum 0% NaCl). The G + C content of its genomic DNA was determined to be 30.5 mol%. The major fatty acid (≥ 10%) components identified included C16:0, C19:0 cyclo ω7c, C19:0 cyclo ω9c, and C18:1 ω9c. The polar lipids were identified as lysophosphatidylethanolamine, phosphatidylethanolamine and glycolipids. The predominant isoprenoid quinones (> 10%) were identified as MK-7, MK-8, MK-9 and MK-10. The amino acid composition of the cell wall was detected as comprising Asp, Glu, Ala, and Lys but the strain lacks meso-diaminopimelic acid. As with other fructophilic lactic acid bacteria, such as L. kunkeei and L. apinorum, strain 10HT was found to prefer d-fructose to d-glucose as a growth substrate. On the basis of these genetic and phenotypic results, the isolate is concluded to represent a novel species, for which the name Lactobacillus kosoi is proposed. The type strain is 10HT (= NBRC 113063T = BCRC 81100T).


Lactobacillus kosoi sp. nov. Kôso Plyphasic taxonomy Fructophilic lactic acid bacteria 16S rRNA gene 



The authors are grateful to the Arsoa R&D Center and the Arsoa Saku Factory for technical supports and sample collection during this study.

Conflict of interest

The authors declare that they have no direct or indirect conflict of interest.

Supplementary material

10482_2018_1019_MOESM1_ESM.docx (866 kb)
Supplementary material 1 (DOCX 867 kb)


  1. Button DK, Schut F, Quang P, Martin R, Robertson BR (1993) Viability and isolation of marine bacteria by dilution culture: theory, procedures, and initial results. Appl Environ Microbiol 59:881–891PubMedPubMedCentralGoogle Scholar
  2. Chiou T-Y, Suda W, Oshima K, Hattori M, Takahashi T (2017) Changes in the bacterial community in the fermentation process of kôso, a Japanese sugar-vegetable fermented beverage. Biosci Biotechnol Biochem 81(2):403–410CrossRefPubMedGoogle Scholar
  3. Collins MD, Jones D (1981) Distribution of isoprenoid quinone structural types in bacteria and their taxonomic implication. Microbiol Rev 45:316–354PubMedPubMedCentralGoogle Scholar
  4. Drucker DB, Megson G, Harty DW, Riba I (1995) Gaskell SJ. Phospholipids of Lactobacillus spp. J Bacteriol 177:6304–6308PubMedGoogle Scholar
  5. Edwards CG, Haag KM, Collins MD, Hutson RA, Huang YC, Haag KM (1998) Lactobacillus kunkeei sp. nov.: a spoilage organism associated with grape juice fermentations. J Appl Microbiol 84:698–702CrossRefPubMedGoogle Scholar
  6. Endo A (2012) Fructophilic lactic acid bacteria inhabit fructose-rich niches in nature. Microb Ecol Health Dis. PubMedPubMedCentralCrossRefGoogle Scholar
  7. Endo A, Irisawa T, Futagawa-Endo Y, Takano K, du Toit M, Okada S, Dicks LM (2012) Characterization and emended description of Lactobacillus kunkeei as a fructophilic lactic acid bacterium. Int J Syst Evol Microbiol 62:500–504CrossRefPubMedGoogle Scholar
  8. Engels V, Georgi T, Wendisch VF (2008) ScrB (Cg2927) is a sucrose-6-phosphate hydrolase essential for sucrose utilization by Corynebacterium glutamicum. FEMS Microbiol Lett 289:80–89CrossRefPubMedGoogle Scholar
  9. Felsenstein J (1981) Evolutionary trees from gene frequencies and quantitative characters: finding maximum likelihood estimates. Evolution 35:1229–1242CrossRefPubMedGoogle Scholar
  10. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefPubMedGoogle Scholar
  11. Kawasaki S, Kurosawa K, Miyazaki M, Sakamoto M, Ohkuma M, Niimura Y (2011) Lactobacillus ozensis sp. nov., isolated from mountain flowers. Int J Syst Evol Microbiol 61:2435–2438CrossRefPubMedGoogle Scholar
  12. Kim M, Oh HS, Park SC, Chun J (2014) Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int J Syst Evol Microbiol 64:346–351CrossRefPubMedGoogle Scholar
  13. Komagata K, Suzuki K (1987) Lipid and cell wall analysis in bacterial systematics. Methods Microbiol 19:161–207CrossRefGoogle Scholar
  14. Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874CrossRefPubMedGoogle Scholar
  15. Lee I, Kim YO, Park SC, Chun J (2016) OrthoANI: an improved algorithm and software for calculating average nucleotide identity. Int J Syst Evol Microbiol 66:1100–1103CrossRefPubMedGoogle Scholar
  16. Liang ZQ, Srinivasan S, Kim YJ, Kim HB, Wang HT, Yang DC (2011) Lactobacillus kimchicus sp. nov., a β-glucosidase-producing bacterium isolated from kimchi. Int J Syst Evol Microbiol 61:894–897CrossRefPubMedGoogle Scholar
  17. Maeno S, Dicks L, Nakagawa J, Endo A (2017) Lactobacillus apinorum belongs to the fructophilic lactic acid bacteria. Biosci Microbiota Food Health 36:147–149CrossRefPubMedPubMedCentralGoogle Scholar
  18. Matsuzaki C, Kamishima K, Matsumoto K, Koga H, Katayama T, Yamamoto K, Hisa K (2014) Immunomodulating activity of exopolysaccharide-producing Leuconostoc mesenteroides strain NTM048 from green peas. J Appl Microbiol 116:980–989CrossRefPubMedGoogle Scholar
  19. Mattarelli P, Holzapfel W, Franz CM, Endo A, Felis GE, Hammes W, Pot B, Dicks L, Dellaglio F (2014) Recommended minimal standards for description of new taxa of the genera Bifidobacterium, Lactobacillus and related genera. Int J Syst Evol Microbiol 64:1434–1451CrossRefPubMedGoogle Scholar
  20. McFrederick QS, Thomas JM, Neff JL, Vuong HQ, Russell KA, Hale AR, Mueller UG (2017) Flowers and wild megachilid bees share microbes. Microb Ecol 73(1):188–200CrossRefPubMedGoogle Scholar
  21. Minnikin DE, Collins MD, Goodfellow M (1979) Fatty acid and polar lipid composition in the classification of Cellulomonas, Oerskovia and related taxa. J Appl Bacteriol 47:87–95. CrossRefGoogle Scholar
  22. Naser SM, Dawyndt P, Hoste B, Gevers D, Vandemeulebroecke K, Cleenwerck I, Vancanneyt M, Swings J (2007) Identification of lactobacilli by pheS and rpoA gene sequence analyses. Int J Syst Evol Microbiol 57:2777–2789CrossRefPubMedGoogle Scholar
  23. Neveling DP, Endo A, Dicks LM (2012) Fructophilic Lactobacillus kunkeei and Lactobacillus brevis isolated from fresh flowers, bees and bee-hives. Curr Microbiol 65:507–515CrossRefPubMedGoogle Scholar
  24. Nishijima M, Araki-Sakai M, Sano H (1997) Identification of isoprenoid quinones by frit-FAB liquid chromatography–mass spectrometry for the chemotaxonomy of microorganisms. J Microbiol Methods 28:113–122CrossRefGoogle Scholar
  25. Olofsson TC, Alsterfjord M, Nilson B, Butler E, Vásquez A (2014) Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov., isolated from the honey stomach of the honeybee Apis mellifera. Int J Syst Evol Microbiol 64:3109–3119CrossRefPubMedPubMedCentralGoogle Scholar
  26. Rosselló-Móra R, Trujillo ME, Sutcliffe IC (2017) Introducing a digital protologue: a timely move towards a database-driven systematics of archaea and bacteria. Antonie Van Leeuwenhoek 110:455–456CrossRefPubMedGoogle Scholar
  27. Rouse S, Harnett D, Vaughan A, van Sinderen D (2008) Lactic acid bacteria with potential to eliminate fungal spoilage in foods. J Appl Microbiol 104:915–923CrossRefPubMedGoogle Scholar
  28. Sasser M (1990) Identification of bacteria by gas chromatography of cellular fatty acids. MIDI Technical Note 101, MIDI Inc., NewarkGoogle Scholar
  29. Schillinger U, Lücke FK (1987) Identification of lactobacilli from meat and meat products. Food Microbiol 4:199–208CrossRefGoogle Scholar
  30. Schleifer KH, Kandler O (1972) Peptidoglycan types of bacterial cell walls and their taxonomic implications. Bacteriol Rev 36:407–477PubMedPubMedCentralGoogle Scholar
  31. Seemann T (2014) Prokka: rapid prokaryotic genome annotation. Bioinformatics 30:2068–2069CrossRefPubMedGoogle Scholar
  32. Simu K, Hagström Å (2004) Oligotrophic bacterioplankton with a novel single-cell life strategy. Appl Environ Microbiol 70:2445–2451CrossRefPubMedPubMedCentralGoogle Scholar
  33. Tamura K, Nei M (1993) Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol Biol Evol 10:512–526PubMedGoogle Scholar
  34. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 11(22):4673–4680CrossRefGoogle Scholar
  35. Yoon SH, Ha SM, Lim JM, Kwon SJ, Chun J (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie Van Leeuwenhoek 110:1281–1286CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing AG, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Biotechnology and Environmental Chemistry, Laboratory of Food Science and TechnologyKitami Institute of TechnologyKitamiJapan
  2. 2.Department of Computational Biology, Graduate School of Frontier SciencesThe University of TokyoKashiwaJapan
  3. 3.Department of Microbiology and ImmunologyKeio University School of MedicineShinjukuJapan
  4. 4.Graduate School of Advanced Science and EngineeringWaseda UniversityShinjukuJapan
  5. 5.Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural UniversityNonoichiJapan
  6. 6.AOB Keioh Group Corporation, ARSOA Research & Development CenterHokutoJapan

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