1887

Abstract

Strain NB2006 was isolated from an isosaccharinate-degrading, nitrate-reducing enrichment culture in minimal freshwater medium at pH 10. Analysis of the 16S rRNA gene sequence indicated that this strain was most closely related to species of the newly established genus Anaerobacillus . This was supported by phenotypic and metabolic characterisation that showed that NB2006 was rod-shaped, Gram-stain-positive, motile and formed endospores. It was an aerotolerant anaerobe and an obligate alkaliphile that grew at pH 8.5–11, could tolerate up to 6 % (w/v) NaCl, and grew at a temperature between 10 and 40 °C. In addition, it could utilise a number of organic substrates, and was able to reduce nitrate and arsenate. The predominant cellular fatty acids were C16 : 0, C16 : 1ω11c, anteiso-C15 : 0, iso-C15 : 0, C16 : 1ω7c/iso-C15 : 0 2-OH and C14 : 0. The cell wall peptidoglycan contained meso-diaminopimelic acid and the DNA G+C content was 37.7 mol%. In silico DNA–DNA hybridization with the four known species of the genus Anaerobacillus showed 21.8, 21.9, 22.4, and 21.5 % relatedness to Anaerobacillus arseniciselenatis DSM 15340, Anaerobacilus alkalidiazotrophicus DSM 22531, Anaerobacillus alkalilacustris DSM 18345, and Anaerobacillus macyae DSM 16346, respectively. NB2006 differed from strains of other species of the genus Anaerobacillus in its ability to metabolise isosaccharinate, an alkaline hydrolysis product of cellulose. On the basis of the consensus of phylogenetic and phenotypic analyses, this strain represents a novel species of the genus Anaerobacillus , for which the name Anaerobacillus isosaccharinicus sp. nov. is proposed. The type strain is NB2006 (=DSM 100644=LMG 30032).

Loading

Article metrics loading...

/content/journal/ijsem/10.1099/ijsem.0.002721
2019-07-02
2019-10-21
Loading full text...

Full text loading...

References

  1. Zavarzina DG, Tourova TP, Kolganova TV, Boulygina ES, Zhilina TN. Description of Anaerobacillus alkalilacustre gen. nov., sp. nov.—Strictly anaerobic diazotrophic bacillus isolated from soda lake and transfer of Bacillus arseniciselenatis, Bacillus macyae, and Bacillus alkalidiazotrophicus to Anaerobacillus as the new combinations A. arseniciselenatis comb. nov., A. macyae comb. nov., and A. alkalidiazotrophicus comb. nov. Microbiology 2009;78: 723– 731 [CrossRef]
    [Google Scholar]
  2. Santini JM, Streimann IC, vanden Hoven RN. Bacillus macyae sp. nov., an arsenate-respiring bacterium isolated from an Australian gold mine. Int J Syst Evol Microbiol 2004;54: 2241– 2244 [CrossRef] [PubMed]
    [Google Scholar]
  3. Sorokin ID, Kravchenko IK, Tourova TP, Kolganova TV, Boulygina ES et al. Bacillus alkalidiazotrophicus sp. nov., a diazotrophic, low salt-tolerant alkaliphile isolated from Mongolian soda soil. Int J Syst Evol Microbiol 2008;58: 2459– 2464 [CrossRef] [PubMed]
    [Google Scholar]
  4. Switzer Blum J, Burns Bindi A, Buzzelli J, Stolz JF, Oremland RS. Bacillus arsenicoselenatis, sp. nov., and Bacillus selenitireducens, sp. nov.: two haloalkaliphiles from Mono Lake, California that respire oxyanions of selenium and arsenic. Arch Microbiol 1998;171: 19– 30 [CrossRef] [PubMed]
    [Google Scholar]
  5. Sjöström E. Wood Chemistry: Fundamentals and Applications San Diego: Academic Press; 1993
    [Google Scholar]
  6. Glaus MA, van Loon LR. Degradation of cellulose under alkaline conditions: new insights from a 12 years degradation study. Environ Sci Technol 2008;42: 2906– 2911 [CrossRef] [PubMed]
    [Google Scholar]
  7. Warwick P, Evans N, Vines S. Studies on some divalent metal α-isosaccharinic acid complexes. Radiochim Acta 2006;94: 363– 368 [CrossRef]
    [Google Scholar]
  8. Keith-Roach MJ. The speciation, stability, solubility and biodegradation of organic co-contaminant radionuclide complexes: a review. Sci Total Environ 2008;396: 1– 11 [CrossRef] [PubMed]
    [Google Scholar]
  9. Kuippers G, Bassil NM, Boothman C, Bryan N, Lloyd JR. Microbial degradation of isosaccharinic acid under conditions representative for the far field of radioactive waste disposal facilities. Mineral Mag 2015;79: 1443– 1454 [CrossRef]
    [Google Scholar]
  10. Kyeremeh IA, Charles CJ, Rout SP, Laws AP, Humphreys PN. Microbial community evolution is significantly impacted by the use of calcium isosaccharinic acid as an analogue for the products of alkaline cellulose degradation. PLoS One 2016;11: e0165832 [CrossRef] [PubMed]
    [Google Scholar]
  11. Charles CJ, Rout SP, Garratt EJ, Patel K, Laws AP et al. The enrichment of an alkaliphilic biofilm consortia capable of the anaerobic degradation of isosaccharinic acid from cellulosic materials incubated within an anthropogenic, hyperalkaline environment. FEMS Microbiol Ecol 2015;91: fiv085 [CrossRef] [PubMed]
    [Google Scholar]
  12. Strand SE, Dykes J, Chiang V. Aerobic microbial degradation of glucoisosaccharinic acid. Appl Environ Microbiol 1984;47: 268– 271 [PubMed]
    [Google Scholar]
  13. Bailey M. Utilization of glucoisosaccharinic acid by a bacterial isolate unable to metabolize glucose. Appl Microbiol Biotechnol 1986;1206: 493– 498
    [Google Scholar]
  14. Bassil NM, Bryan N, Lloyd JR. Microbial degradation of isosaccharinic acid at high pH. Isme J 2015;9: 310– 320 [CrossRef] [PubMed]
    [Google Scholar]
  15. Lovley DR, Greening RC, Ferry JG. Rapidly growing rumen methanogenic organism that synthesizes coenzyme M and has a high affinity for formate. Appl Environ Microbiol 1984;48: 81– 87 [PubMed]
    [Google Scholar]
  16. Rout SP, Charles CJ, Garratt EJ, Laws AP, Gunn J et al. Evidence of the generation of isosaccharinic acids and their subsequent degradation by local microbial consortia within hyper-alkaline contaminated soils, with relevance to intermediate level radioactive waste disposal. PLoS One 2015;10: e0119164 [CrossRef] [PubMed]
    [Google Scholar]
  17. Rizoulis A, Steele HM, Morris K, Lloyd JR. The potential impact of anaerobic microbial metabolism during the geological disposal of intermediate-level waste. Mineral Mag 2012;76: 3261– 3270 [CrossRef]
    [Google Scholar]
  18. Lane DJ. 16S/23S rRNA sequencing. In Stackebrandt E, Goodfellow M. (editors) Nucleic Acid Techniques in Bacterial Systematics London: John Wiley & Sons Ltd; 1991; pp. 115– 174
    [Google Scholar]
  19. Handley KM, Héry M, Lloyd JR. Marinobacter santoriniensis sp. nov., an arsenate-respiring and arsenite-oxidizing bacterium isolated from hydrothermal sediment. Int J Syst Evol Microbiol 2009;59: 886– 892 [CrossRef] [PubMed]
    [Google Scholar]
  20. Yoon SH, Ha SM, Kwon S, Lim J, Kim Y et al. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int J Syst Evol Microbiol 2017;67: 1613– 1617 [CrossRef] [PubMed]
    [Google Scholar]
  21. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 1994;22: 4673– 4680 [CrossRef] [PubMed]
    [Google Scholar]
  22. Kumar S, Stecher G, Tamura K. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 2016;33: 1870– 1874 [CrossRef] [PubMed]
    [Google Scholar]
  23. Saitou N, Nei M. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4: 406– 425 [CrossRef] [PubMed]
    [Google Scholar]
  24. Tamura K, Nei M, Kumar S. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 2004;101: 11030– 11035 [CrossRef] [PubMed]
    [Google Scholar]
  25. Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 1980;16: 111– 120 [CrossRef] [PubMed]
    [Google Scholar]
  26. Felsenstein J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 1985;39: 783– 791 [CrossRef] [PubMed]
    [Google Scholar]
  27. Gault AG, Jana J, Chakraborty S, Mukherjee P, Sarkar M et al. Preservation strategies for inorganic arsenic species in high iron, low-Eh groundwater from West Bengal, India. Anal Bioanal Chem 2005;381: 347– 353 [CrossRef] [PubMed]
    [Google Scholar]
  28. Wang JP, Liu B, Liu GH, Ge CB, Chen QQ et al. Genome sequence of Anaerobacillus macyae JMM-4T (DSM 16346), the first genomic information of the newly established genus Anaerobacillus. Genome Announc 2015;3: e00922-15 [CrossRef] [PubMed]
    [Google Scholar]
  29. Bassil NM, Lloyd JR. Draft genome sequences of four alkaliphilic bacteria belonging to the Anaerobacillus genus. Genome Announc 2017;5: e01493-16 [CrossRef] [PubMed]
    [Google Scholar]
  30. Lee I, Ouk Kim Y, Park SC, Chun J. OrthoANI: An improved algorithm and software for calculating average nucleotide identity. Int J Syst Evol Microbiol 2016;66: 1100– 1103 [CrossRef] [PubMed]
    [Google Scholar]
  31. Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M. Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 2013;14: 60 [CrossRef] [PubMed]
    [Google Scholar]
  32. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014;30: 2068– 2069 [CrossRef] [PubMed]
    [Google Scholar]
  33. Page AJ, Cummins CA, Hunt M, Wong VK, Reuter S et al. Roary: rapid large-scale prokaryote pan genome analysis. Bioinformatics 2015;31: 3691– 3693 [CrossRef] [PubMed]
    [Google Scholar]
  34. Chan JZ, Halachev MR, Loman NJ, Constantinidou C, Pallen MJ. Defining bacterial species in the genomic era: insights from the genus Acinetobacter. BMC Microbiol 2012;12: 302 [CrossRef] [PubMed]
    [Google Scholar]
  35. Wayne LG, Moore WEC, Stackebrandt E, Kandler O, Colwell RR et al. Report of the ad hoc committee on reconciliation of approaches to bacterial systematics. Int J Syst Evol Microbiol 1987;37: 463– 464 [CrossRef]
    [Google Scholar]
  36. Meier-Kolthoff JP, Göker M, Spröer C, Klenk HP. When should a DDH experiment be mandatory in microbial taxonomy?. Arch Microbiol 2013;195: 413– 418 [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/ijsem/10.1099/ijsem.0.002721
Loading
/content/journal/ijsem/10.1099/ijsem.0.002721
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error