Advertisement

Current Microbiology

, Volume 64, Issue 4, pp 343–348 | Cite as

Evidence for Two Putative Holin-Like Peptides Encoding Genes of Bacillus pumilus Strain WAPB4

  • Ratchaneewan Aunpad
  • Watanalai Panbangred
Article

Abstract

An open reading frame encoding a 71-amino acid BhlA bacteriocin-related holin-like peptide was present upstream of 86-amino acid holin-like peptide, xhlB, encoding gene in the genome of Bacillus pumilus strain WAPB4. Analysis of BhlA using TMHMM server suggested one putative transmembrane domain at the N-terminal part and a number of highly charged amino acid residues at the C-terminal part. XhlB of B. pumilus strain WAPB4 composed of two putative transmembrane domains separated by a β-turn, and numerous charged residues in the C-terminus. The dual start motifs were found in both BhlA and XhlB. Structural analysis of their sequence revealed features characteristic for holin. To analyze the effect of BhlA on bacteria cell, its ORF was cloned and expressed in Escherichia coli BL21(DE3). Expression of holin-like peptide, BhlA, was found to be toxic to the host cell. The site of action of BhlA is on the cell membrane and caused bacterial death by cell membrane disruption as clearly demonstrated by transmission electron microscopy or TEM.

Keywords

Endolysin Common Structural Feature Charge Amino Acid Residue Putative Transmembrane Domain Cell Membrane Disruption 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was supported by a research grant for new scholars from the Commission on Higher Education in collaboration with the Thailand Research Fund and partially supported by the National Research Council of Thailand.

References

  1. 1.
    Abriouel H, Franz CM, Omar NB, Gálvez A (2011) Diversity and applications of Bacillus bacteriocins. FEMS Microbiol Rev 35:201–232PubMedCrossRefGoogle Scholar
  2. 2.
    Aunpad R, Na-Bangchang K (2007) Pumilicin 4, a novel bacteriocin with anti-MRSA and anti-VRE activity produced by newly isolated bacteria Bacillus pumilus strain WAPB4. Curr Micobiol 55:308–313CrossRefGoogle Scholar
  3. 3.
    Anthony T, Stalin CG, Rajesh T, Gunasekaran P (2010) Functional analysis of a putative holin-like peptide coding gene in the genome of Bacillus licheniformis AnBa9. Arch Microbiol 192:51–56PubMedCrossRefGoogle Scholar
  4. 4.
    Delisle AL, Gerard JB, Guo M (2006) Isolation and expression of the lysis genes of Actinomyces naeslundii phage Av-1. Appl Environ Microbiol 72:1110–1117PubMedCrossRefGoogle Scholar
  5. 5.
    Horii T, Suzuki Y, Kobayashi M (2002) Characterization of a holin (HolNU3–1) in methicillin-resistant Staphylococcus aureus host. FEMS Immunol Med Microbiol 34(4):307–310PubMedCrossRefGoogle Scholar
  6. 6.
    Kyogoku K, Sekiguchi J (1999) Cloning and sequencing of a new holin-encoding gene of Bacillus licheniformis. Gene 168:61–65CrossRefGoogle Scholar
  7. 7.
    Oki M, Kakikawa M, Nakamura S, Yamamura ET, Watanabe K, Sasamoto M, Taketo A, Kodaira KI (1997) Functional and structural features of the holin HOL protein of the Lactobacillus plantarum phage φg1e: analysis in Escherichia coli system. Gene 197:137–145PubMedCrossRefGoogle Scholar
  8. 8.
    Rodas P, Trombert AN, Mora GC (2011) A holin remnant protein encoded by STY1365 is involved in envelope stability of Salmonella enterica serovar Typhi. FEMS Microbiol Lett 321(1):58–66PubMedCrossRefGoogle Scholar
  9. 9.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring HarborGoogle Scholar
  10. 10.
    Schagger H, von Jagow G (1987) Tricine-sodium dodecyl sulphate-polyacrylamide gel electrophoresis for the separation of proteins in the range of 1 to 100 kDa. Anal Biochem 166:368–379PubMedCrossRefGoogle Scholar
  11. 11.
    Sheehan MM, Stanley E, Fitzgerald GF, Van Sinderen D (1999) Identification and characterization of a lysis module present in a large proportion of bacteriophages infecting Streptococcus thermophilus. Appl Environ Microbiol 65:569–577PubMedGoogle Scholar
  12. 12.
    Tan KS, Wee BY, Song KP (2001) Evidence for holin function of tcdE gene in the pathogenicity of Clostridium difficile. J Med Microbiol 50:613–619PubMedGoogle Scholar
  13. 13.
    Young RY (1992) Bacteriophage lysis: mechanism and regulation. Microbiol Rev 56:430–481PubMedGoogle Scholar
  14. 14.
    Young R, Bläsi U (1995) Holins: form and function in bacteriophage lysis. FEMS Microbiol Rev 17:191–205PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Graduate Program in Biomedical Sciences, Faculty of Allied Health SciencesThammasat UniversityPathumthaiThailand
  2. 2.Department of Biotechnology, Faculty of ScienceMahidol UniversityBangkokThailand

Personalised recommendations