Skip to main content
Log in

Clostridium swellfunianum sp. nov., a novel anaerobic bacterium isolated from the pit mud of Chinese Luzhou-flavor liquor production

  • Original Paper
  • Published:
Antonie van Leeuwenhoek Aims and scope Submit manuscript

Abstract

A novel Gram-positive, strictly anaerobic, spore-forming, rod-shaped bacterium, designated strain S11-3-10T, was isolated from the pit mud used for Chinese Luzhou-flavor liquor production. Phylogenetic analysis based on 16S rRNA gene sequencing revealed that the strain formed a monophyletic clade with the closely related type strains of Clostridium cluster I and was most closely related to Clostridium amylolyticum JCM 14823T (94.38 %). The temperature, pH, and NaCl range for growth was determined to be 20–45 °C (optimum 37 °C), 4.0–10.0 (optimum pH 7.3), and 0–3.0 % (w/v), respectively. The strain was able to tolerate up to 7.5 % (v/v) ethanol. Yeast extract or peptone was found to be required for growth. Acids were found to be produced from glucose, mannose and trehalose. The major end products from glucose fermentation were identified as ethanol, acetate and hydrogen. The polar lipids were found to consist of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and unidentified phospholipids and polar lipids. The major fatty acids (>5 %) were identified as iso-C15:0, C16:0, C16:0 dma, C14:0, anteiso-C15:0 and iso-C13:0. No respiratory quinone was detected. The diamino acid in the cell wall peptidoglycan was identified as meso-diaminopimelic acid and the whole-cell sugars were found to include galactose and glucose as major components. The DNA G+C content was determined to be 36.4 mol%. Based on the phylogenetic, chemotaxonomic and phenotypic evidence, the isolate is considered to represent a novel species of the genus Clostridium for which the name Clostridium swellfunianum sp. nov. is proposed. The type strain is S11-3-10T (=DSM 27788T = JCM 19606T = CICC 10730T).

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Andreesen JR, Zindel U, Durre P (1985) Clostridium cylindrosporum (ex Barker and Beck 1942) nom.rev. Int J Syst Bacteriol 35:206–208

    Article  Google Scholar 

  • Andreesen JR, Bahl H, Gottschalk G (1989) Introduction to the physiology and biochemistry of the genus Clostridium. In: Minton NP, Clarke DJ (eds) Clostridia (Biotechnology Handbooks). Plenum Publishing corporation, New York, pp 27–53

    Google Scholar 

  • Bornstein B, Barker H (1948) The nutrition of Clostridium kluyveri. J Bacteriol 55:223–230

    CAS  PubMed Central  PubMed  Google Scholar 

  • Collins M, Jones D (1980) Lipids in the classification and identification of coryneform bacteria containing peptidoglycans based on 2,4-diaminobutyric acid. J Appl Microbiol 48:459–470

    CAS  Google Scholar 

  • Collins MD, Lawson PA, Willems A, Cordoba JJ, Fernandez-Garayzabal J, Garcia P, Cai J, Hippe H, Farrow JAE (1994) The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol 44:812–826

    Article  CAS  PubMed  Google Scholar 

  • Cummins CS, Johnson JL (1971) Taxonomy of the clostridia: wall composition and DNA homologies in Clostridium butyricum and other butyric acid-producing clostridia. J Gen Microbiol 67:33–46

    Article  Google Scholar 

  • Ding XF, Wu CD, Zhang LQ, Zheng J, Zhou RQ (2013) Characterization of eubacterial and archaeal community diversity in the pit mud of Chinese Luzhou-flavor liquor by nested PCR–DGGE. World J Microbiol Biotechnol 30:605–612

    Article  PubMed  Google Scholar 

  • Dong XZ, Cai MY (2001) Determinative manual for routine bacteriology. Scientific Press, Beijing (English translation)

    Google Scholar 

  • Fan WL, Qian MC (2006) Characterization of aroma compounds of chinese “Wuliangye” and “Jiannanchun” liquors by aroma extract dilution analysis. J Agric Food Chem 54:2695–2704

    Article  CAS  PubMed  Google Scholar 

  • Hasegawa T, Takizawa M, Tanida S (1983) A rapid analysis for chemical grouping of aerobic actinomycetes. J Gen Appl Microbiol 29:319–322

    Article  CAS  Google Scholar 

  • Huang Y, Sun YJ, Ma SC, Chen L, Zhang H, Deng Y (2013) Isolation and characterization of Keratinibaculum paraultunense gen. nov., sp. nov., a novel thermophilic, anaerobic bacterium with keratinolytic activity. FEMS Microbiol Lett 345:56–63

    Article  CAS  PubMed  Google Scholar 

  • Hungate R (1969) A roll-tube method for cultivation of strict anaerobes. Methods Microbiol 3B:117–132

    Article  CAS  Google Scholar 

  • Johnston NC, Goldifine H (1983) Lipid composition in the classification of the butyric acid-producing clostridia. J Gen Microbiol 129:1075–1081

    CAS  PubMed  Google Scholar 

  • Johnston NC, Sunar SA, Guan ZQ, Ribeiro AA, Daldal F, Raetz CRH, Goldifine H (2010) A phosphoethanolamine-modified glycosyl diradylglycerol in the polar lipids of clostridium tetani. J Lipid Res 51:1953–1961

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Karita S, Nakayama K, Goto M, Sakka K, Kim WJ, Ogawa S (2003) A novel cellulolytic, anaerobic, and thermophilic bacterium, Moorella sp. strain F21. Biosci Biotechnol Biochem 67:183–185

    Article  CAS  PubMed  Google Scholar 

  • Kim OS, Cho YJ, Lee K, Yoon SH, Kim M, Na H, Park SC, Jeon YS, Lee JH, Yi H, Won S, Chun J (2012) Introducing EzTaxon-e, a prokaryotic 16S rRNA gene sequence database with phylotypes that represent uncultured specied. Int J Syst Evol Microbiol 62:716–721

    Article  CAS  PubMed  Google Scholar 

  • Kimberly SB, Rachael ED, Fred AR, William MM (2010) Clostridium hydrogeniformans sp. nov. and Clostridium cavendishii sp. nov., hydrogen-producing bacteria from chlorinated solvent-contaminated groundwater. Int J Syst Evol Microbiol 60:358–363

    Article  Google Scholar 

  • Mesbah M, Premachandran U, Whitman WB (1989) Precise measurement of the G+C content of deoxyribonucleic acid by high-performance liquid chromatography. Int J Syst Bacteriol 39:159–167

    Article  CAS  Google Scholar 

  • Minnikin DE, O’Donnell AG, Goodfellow M, Alderson G, Athalye M, Schaal A, Parlett JH (1984) An integrated procedure for the extraction of bacterial isoprenoid quinines and polar lipids. J Microbiol Methods 2:233–241

    Article  CAS  Google Scholar 

  • Shin SG, Lee S, Lee C, Hwang K, Hwang S (2010) Qualitative and quantitative assessment of microbial community in batch anaerobic digestion of secondary sludge. Bioresour Technol 101:9461–9470

    Article  CAS  PubMed  Google Scholar 

  • Smibert RM, Krieg NR (1994) Phenotypic characterization. In: Gerhardt P, Murray RGE, Wood WA, Krieg NR (eds) Methods for general and molecular bacteriology. American Society for Microbiology, Washington, pp 607–654

    Google Scholar 

  • Song L, Dong XZ (2008) Clostridium amylolyticum sp. nov., isolated from H2-producing UASB granules. Int J Syst Evol Microbiol 58:2132–2135

    Article  CAS  PubMed  Google Scholar 

  • Spring S, Merkhoffer B, Weiss N, Kroppenstedt RM, Hippe H, Stackebrandt E (2003) Characterization of novel psychrophilic clostridia from an Antarctic microbial mat: description of Clostridium frigoris sp. nov., Clostridium lacusfryxellense sp. nov., Clostridium bowmanii sp. nov. and Clostridium psychrophilum sp. nov. and reclassification of Clostridium laramiense as Clostridium estertheticum subsp. Laramiense subsp. nov. Int J Syst Evol Microbiol 53:1019–1029

    Article  CAS  PubMed  Google Scholar 

  • Stackebrandt E, Goebel BM (1994) Taxonomic note: a place for DNA–DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. Int J Syst Bacteriol 44:846–849

    Article  CAS  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) Mega 5: molecular evolutionary genetics analysis using maximum likelihood evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Tao Y, Li JB, Rui JP, Xu ZC, Zhou Y, Hu XH, Wang X, Liu MH, Li DP, Li XZ (2014) Prokaryotic communities in pit mud from different-aged cellars used for the production of Chinese strong-flavor liquor. Appl Environ Microbiol 80:2254–2260

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • 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 Acid Res 22:4673–4680

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Worden R, Grethlein A, Jain M, Datta R (1991) Production of butanol and ethanol from synthesis gas via fermentation. Fuel 70:615–619

    Article  CAS  Google Scholar 

  • Wu YY, Xue TR, Chen SR, Lu SY, Liu AH, Lin S, Tang WY, Tang HJ, Zhao X (1991) Study on distribution and action of anerobic bacteria in Wuliang old fermentation pits. Acta Microbiogica Sinica 31:299–307 (in Chinese)

    Google Scholar 

  • Zhang WX, Qiao ZW, Shigemastsu T, Tang YQ, Hu C, Morimura S, Kida K (2005) Analysis of the bacterial community in Zaopei during production of Chinese Luzhou-flavor liquor. J Inst Brew 111:215–222

    Article  CAS  Google Scholar 

  • Zheng J, Liang R, Zhang LQ, Wu CD, Zhou R, Liao X (2013) Characterization of microbial communities in strong aromatic liquor fermentation pit muds of different ages assessed by combined DGGE and PLFA analyses. Food Res Int 1:660–666

    Article  Google Scholar 

Download references

Acknowledgments

This work was financially supported by Natural Science Foundation of China (NSFC No. 31071585).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Wenxue Zhang.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 13671 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, C., Huang, D., Liu, L. et al. Clostridium swellfunianum sp. nov., a novel anaerobic bacterium isolated from the pit mud of Chinese Luzhou-flavor liquor production. Antonie van Leeuwenhoek 106, 817–825 (2014). https://doi.org/10.1007/s10482-014-0251-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10482-014-0251-z

Keywords

Navigation