Skip to main content
SpringerLink
Log in

Role of stereospecific nature of germinants in Bacillus megaterium spores germination

  • Original Article
  • Published:
3 Biotech Aims and scope Submit manuscript

Abstract

The present study was undertaken with the objective to assess the effect of distinct stereoisomeric forms of nutrient germinants (selected sugars and amino acids) on the process of germination onset in dormant spores of Bacillus megaterium MTCC 2949. In this respect, epimers of glucose and enantiomers of alanine were employed in current work. When supplemented with these stereoisomers, spores were found germinated only with d-glucose and d-mannose among epimers of glucose and only with l-alanine among enantiomers of alanine. Interestingly, germination in spores was observed to negligible extent with d-galactose and d-alanine. These findings were obtained on the basis of four type of germination assays, namely reduction in absorbance measured at 600 nm (≤5 to ≥30%), refractility examination (phase bright and dark), esterase assay [fluorescence units 0.455–94.62 (×103)] and fluorescent staining (fluorescent/non-fluorescent signals). Understanding of spores germination process and efficacy of different forms of germinants to trigger germination is of immense importance. It aids in development of sensing and sterilization indicating tools employing chiefly spores as biorecognition elements and in uncovering the mechanism of diseases, food contamination and spoilages resulting from the germination of spores. The findings of current work support the possibility to explore such germination mechanism by significantly giving the clue for potential existence of stereospecific receptor sites on the surface of B. megaterium spores. Perhaps, these sites can specifically differentiate and recognize stereoisomerically diverse forms of germinants for induction of germination.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Alberto F, Broussolle V, Mason DR, Carlin F, Peck MW (2003) Variability in spore germination response by strains of proteolytic Clostridium botulinum types A, B and F. Lett Appl Microbiol 36:41–45

    Article  CAS  Google Scholar 

  • Ando Y (1971) The germination requirements of spores of Clostridium botulinum type E Japan. J Microbiol 15:515–525

    CAS  Google Scholar 

  • Barlass PJ, Houston CW, Clements MO, Moir A (2002) Germination of Bacillus cereus spores in response to l-alanine and to inosine: the roles of gerL and gerQ operons. Microbiology 148:2089–2095. doi:10.1099/00221287-148-7-2089

    Article  CAS  Google Scholar 

  • Black EP, Setlow P, Hocking AD, Stewart CM, Kelly AL, Hoover DG (2007) Response of spores to high-pressure processing. Comp Rev Food Sci Food Saf 6:103–119

    Article  CAS  Google Scholar 

  • Borch-Pedersen K, Lindbäck T, Madslien EH, Kidd SW, O’Sullivan K, Granum PE, Aspholm M (2016) The cooperative and interdependent roles of GerA, GerK, and Ynd in germination of Bacillus licheniformis spores. Appl Environ Microbiol 82:4279–4287

    Article  CAS  Google Scholar 

  • Broussolle V, Alberto F, Shearman CA, Mason DR, Botella L, Nguyen-The C, Peck MW, Carlin F (2002) Molecular and physiological characterization of spore germination in Clostridium botulinum and C. sporogenes. Anaerobe 8:89–100

    Article  CAS  Google Scholar 

  • Cabrera-Martinez RM, Tovar-Rojo F, Vepachedu VR, Setlow P (2003) Effects of overexpression of nutrient receptors on germination of spores of Bacillus subtilis. J Bacteriol 185:2457–2464. doi:10.1128/JB.185.8.2457-2464.2003

    Article  CAS  Google Scholar 

  • Cava F, Lam H, de Pedro MA, Waldor MK (2011) Emerging knowledge of regulatory roles of d-amino acids in bacteria. Cell Mol Life Sci 68:817–831

    Article  CAS  Google Scholar 

  • Christie G, Lowe CR (2007) Role of chromosomal and plasmid-borne receptor homologues in the response of Bacillus megaterium QM B1551 spores to germinants. J Bacteriol 189:4375–4383. doi:10.1128/JB.00110-07

    Article  CAS  Google Scholar 

  • Downes FP, Ito K (eds) (2001) Compendium of methods for the microbiological examination of foods, 4th edn. American Public Health Association, Washington, DC, USA

  • Ferencko L, Cote MA, Rotman B (2004) Esterase activity as a novel parameter of spore germination in Bacillus anthracis. Biochem Biophys Res Commun 319:854–858. doi:10.1016/j.bbrc.2004.05.062

    Article  CAS  Google Scholar 

  • Fisher N, Hanna P (2005) Characterization of Bacillus anthracis germinant receptors in vitro. J Bacteriol 187:8055–8062

    Article  CAS  Google Scholar 

  • Ghosh S (2013) Understanding the mechanism of Bacillus subtilis spore germination. Doctoral Dissertation, University of Connecticut Graduate School

  • Hashimoto T, Frieben WR, Conti SF (1969) Germination of single bacterial spores. J Bacteriol 98:1011–1020

    CAS  Google Scholar 

  • Hitchins AD, Kahn AJ, Slepecky RA (1968) Interference contrast and phase contrast microscopy of sporulation and germination of Bacillus megaterium. J Bacteriol 96:1811–1817

    CAS  Google Scholar 

  • Hornstra LM, de Vries YP, Wells-Bennik MHJ, de Vos WM, Abee T (2006) Characterization of germination receptors of Bacillus cereus ATCC 14579. Appl Environ Microbiol 72:44–53. doi:10.1128/AEM

    Article  CAS  Google Scholar 

  • Hyatt MT, Levinson HS (1964) Effect of sugars and other carbon compounds on germination and postgerminative development of Bacillus megaterium spores. J Bacteriol 88:1403–1415

    CAS  Google Scholar 

  • Igarashi T, Setlow B, Paidhungat M, Setlow P (2004) Effects of a gerF (lgt) mutation on the germination of spores of Bacillus subtilis. J Bacteriol 186:2984–2991. doi:10.1128/JB.186.10.2984-2991.2004

    Article  CAS  Google Scholar 

  • Kristin CSR (2012) Diversity in germination response among Bacillus licheniformis strains. Master thesis, Norwegian University of Life Sciences

  • Kumar N, Singh NA, Singh VK, Bhand S, Malik RK (2010) Development of spore inhibition based-enzyme substrate assay (SIB-ESA) for monitoring aflatoxin M1 in milk. Indian Patent Reg. No. 3064/DEL/2010: Publication No. 46/2012

  • Løvdal IS, From C, Madslien EH, Romundset KCS, Klufterud E, Rosnes JT, Granum PE (2012) Role of the gerA operon in l-alanine germination of Bacillus licheniformis spores. BMC Microbiol 12:1–12. doi:10.1186/1471-2180-12-34

    Article  Google Scholar 

  • Madslien EH, Granum PE, Blatny JM, Lindbäck T (2014) L-alanine-induced germination in Bacillus licheniformis the impact of native gerA sequences. BMC Microbiol 14:101–110. doi:10.1186/1471-2180-14-101

    Article  Google Scholar 

  • McCann KP, Robinson C, Sammons RL, Smith DA, Corfe BM (1996) Alanine germination receptors of Bacillus subtilis. Lett Appl Microbiol 23:290–294

    Article  CAS  Google Scholar 

  • Moir A (2006) How do spores germinate? J Appl Microbiol 101:526–530. doi:10.1111/j.1365-2672.2006.02885.x

    Article  CAS  Google Scholar 

  • Moir A, Corfe BM, Behravan J (2002) Spore germination. Cell Mol Life Sci 59:403–409. doi:10.1007/s00018-002-8432-8

    Article  CAS  Google Scholar 

  • Nicholson W (2002) Roles of Bacillus endospores in the environment. Cell Mol Life Sci 59:410–416. doi:10.1007/s00018-002-8433-7

    Article  CAS  Google Scholar 

  • Paidhungat M, Setlow P (2002) Spore germination and outgrowth. In: Sonenshein AL, Hoch JA, Losick R (eds) Bacillus subtilis and its closest relatives: from genes to cells. American Society for Microbiology, Washington

    Google Scholar 

  • Paredes-Sabja D, Torres JA, Setlow P, Sarker MR (2008) Clostridium perfringens spore germination: characterization of germinants and their receptors. J Bacteriol 190:1190–1201

    Article  CAS  Google Scholar 

  • Paredes-Sabja D, Setlow P, Sarker MR (2011) Germination of spores of Bacillales and Clostridiales species: mechanisms and proteins involved. Trends Microbiol 19:85–94. doi:10.1016/j.tim.2010.10.004

    Article  CAS  Google Scholar 

  • Racine FM, Dills SS, Vary JC (1979) Glucose-triggered germination of Bacillus megaterium spores. J Bacteriol 138:442–445

    CAS  Google Scholar 

  • Ramirez N, Abel-Santos E (2010) Requirements for germination of Clostridium sordellii spores in vitro. J Bacteriol 192:418–425

    Article  CAS  Google Scholar 

  • Ross C, Abel-Santos E (2010) The ger receptor family from sporulating bacteria. Curr Issues Mol Biol 12:47–158

    Google Scholar 

  • Sacks LE, Thompson PA (1971) Germination requirements of Bacillus macerans spores. J Bacteriol 105:739–746

    CAS  Google Scholar 

  • Setlow P (2003) Spore germination—review article. Curr Opin Microbiol 6:550–556

    Article  CAS  Google Scholar 

  • Setlow P (2013) Summer meeting 2013—when the sleepers wake: the germination of spores of Bacillus species. J Appl Microbiol 115:1251–1268. doi:10.1111/jam.12343

    Article  CAS  Google Scholar 

  • Setlow P, Johnson EA (2012) Spores and their significance. In: Doyle MP, Buchanan RL (eds) Food microbiology: fundamentals and frontiers, 4th edn. ASM Press, Washington

    Google Scholar 

  • Setlow B, Peng L, Loshon CA, Li YQ, Christie G, Setlow P (2009) Characterization of the germination of Bacillus megaterium spores lacking enzymes that degrade the spore cortex. J Appl Microbiol 107:318–328. doi:10.1111/j.1365-2672.2009.04210.x

    Article  CAS  Google Scholar 

  • Thakur G, Yadav A, Tehri N, Kumar N, Raghu HV, Singh N, Singh VK (2013) Rapid & novel microscopy technique to detect germination initiation and specificity in Bacillus spores. Int J Res Pure Appl Microbiol 3:134–138

    Google Scholar 

  • Vary J, Halvorson H (1965) Kinetics of germination of Bacillus spores. J Bacteriol 89:1340–1347

    CAS  Google Scholar 

  • Zhang P, Garner W, Yi X, Yu J, Li YQ, Setlow P (2010) Factors affecting variability in time between addition of nutrient germinants and rapid dipicolinic acid release during germination of spores of Bacillus species. J Bacteriol 192:3608–3619. doi:10.1128/JB.00345-10

    Article  CAS  Google Scholar 

  • Zuberi AR, Moir A, Feavers IM (1987) The nucleotide sequence and gene organization of the gerA spore germination operon of Bacillus subtilis 168. Gene 51:1–11

    Article  CAS  Google Scholar 

Download references

Acknowledgements

Niche Area of Excellence (NAE) is greatly acknowledged for supporting this research work. The Director NDRI is thankfully acknowledged for institute fellowship for pursuing Ph.D. to Nimisha Tehri.

Author information

Authors and Affiliations

  1. Microbial Biosensors and Food Safety Laboratory, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, Haryana, 132001, India

    Nimisha Tehri, Naresh Kumar, H. V. Raghu, Geetika Thakur & Pradip Kumar Sharma

Authors
  1. Nimisha Tehri
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Naresh Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. H. V. Raghu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Geetika Thakur
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pradip Kumar Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Nimisha Tehri.

Ethics declarations

Conflict of interest

Authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tehri, N., Kumar, N., Raghu, H.V. et al. Role of stereospecific nature of germinants in Bacillus megaterium spores germination. 3 Biotech 7, 259 (2017). https://doi.org/10.1007/s13205-017-0897-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13205-017-0897-0

Keywords

Search

Navigation