Malaysian Journal of Analytical Sciences
Vol 19 No 4 (2015): 831 - 840
BACTERIOCIN ISOLATED FROM HALOMONAS
SP.: A BACTERIAL DING PROTEIN?
(Pemencilan Bakteriosin dari Halomonas
sp. : Sejenis Bakteria DING protin?)
Atirah Azemin1*, Peter
Klappa2, Mohd Shahir Shamsir Omar1
1Faculty of Biosciences and
Medical Engineering,
Universiti Teknologi Malaysia,81310 Skudai, Johor,
Malaysia
2School of Biosciences, Stacey
Building,
University of Kent, Canterbury, Kent, CT2 7NJ,United
Kingdom
*Corresponding author: atirahasyifa@yahoo.com
Received: 23
November 2014; Accepted: 27 June 2015
Abstract
A marine halophile, Halomonas sp. strain M3 was
isolated from Straits of Johor, Malaysia and produce bacteriocin CC that acts
as bacteriostatic agent. Characterisation of the bacterium showed that optimal
growth and bacteriocin production is at ambient temperature, pH of 8-8.5 in
nutrient broth medium supplemented with 2.9% w/v NaCI to mimic
saltwater conditions. The stability studies indicated that bacteriocin CC is
heat-labile (35°C-50°C) and was stable over 2 years when stored in 0.02M
Tris-HCI with 30-60% glycerol at 4°C. A loss of activity was detected after
proteolytic enzymes treatment, indicating the proteinaceous nature of the
antimicrobial compound. The amino acid sequence of bacteriocin CC was obtained
by Edman degradation and MALDI-TOF analysis, showed the characteristic sequence
of a DING protein (D-I-N-G-G-G-A-T-L-P-Q-A-L-Y-Q) in size 38.9-kDa at pI of 6.8. These proteins constitute a
conserved and widely distributed set of proteins found in all kingdoms with
ligand-binding activities and hydrolytic enzyme, suggesting a possible role in
cell signalling and bio mineralization in DING isolates. Intriguingly, DING
proteins also have been involved as an anti-tumour agent in humans. Thus,
bacteriocin CC as DING protein family members should be further studied to
investigate its potential as a novel antimicrobial agent.
Keywords: antimicrobial activity, bacteriocin,
DING protein, Halomonas sp. Strain M3
Abstrak
Sejenis halofil marin Halomonas sp. strain M3 telah
dipencilkan daripada Selat Johor, Malaysia dan menghasilkan bakteriosin CC yang
bertindak sebagai ejen bakteriostatik. Pencirian bakteria ini menunjukkan
bahawa pertumbuhan yang optima pada suhu bilik, pH 8-8.5 dalam nutrien media yang
ditambah dengan 2.9% w/v NaCI untuk meniru keadaan air masin. Kajian kestabilan menunjukkan
bahawa bakteriosin CC adalah tahan haba (35°C-50
°C) dan stabil sepanjang 2 tahun apabila disimpan dalam 0.02M Tris-HCI bersama 30-60%
gliserol pada 4°C. Kehilangan aktiviti telah dikesan selepas rawatan enzim proteolitik, menunjukkan sifat
protin dalam komposisi antimikrob. Urutan asid amino bakteriosin CC telah
diperolehi dengan menggunakan degradasi Edman dan analisis MALDI-TOF, menunjukkan
ciri protin DING (D-I-N-G-G-G-A-T-L-P-Q-A-L-Y-Q) dalam saiz 38.9-kDa pada pI
6.8. Protin ini membentuk satu set yang terpelihara dan edaran protin ini didapati
secara meluas dalam semua hidupan dengan aktiviti pengikatan-ligan dan enzim
hidrolitik, mencadangkan peranan yang mungkin ada dalam pengisyaratan sel dan
bio mineral dalam pemencilan DING yang lain. Menariknya, DING protin juga terlibat
sebagai agen anti-tumor pada manusia. Oleh itu, bakteriosin CC sebagai ahli keluarga
protin DING, perlu dikaji lagi untuk menyiasat sekiranya ia berpotensi sebagai
ejen antimikrob yang baru.
Kata kunci: aktiviti antimikrob, bakteriosin,
DING protin, Halomonas sp. Strain M3
References
1. Ngo, D-H., Vo, T-S., Ngo, D-N., Wijesekara,
I. and Kim, S-K. (2012). Biological activities and potential health benefits of
bioactive peptides derived from marine organisms. Biological Macromolecules,
51: 378-383.
2. Faulkner, D.J. (2000). Marine natural
products. Natural Product Reports, 17: 7-55.
3. Chen, L., Wang, G., Bu, T., Zhang, Y.,
Wang, Y., Liu, M. and Lin, X. (2010). Phylogenetic analysis and screening of
antimicrobial and cytotoxic activities of moderately halophilic bacteria
isolated from the Weihai Solar Saltern (China). World Microbiology Biotechnology,
26: 879-888.
4. Pašić, L., Velikonja, B.. H.
and Ulrih, N. P. (2008). Optimization of the culture conditions for the
production of a bacteriocin from halophilic archaeon Sech7a. Preparative
Biochemistry Biotechnology, 38: 229-245.
5. Ishibashi, M., Yamashita, S. and Tokunaga,
M. (2005). Characterization of halophilic alkaline phosphatase from Halomonas
sp. 593, a moderately halophilic bacterium. Biosciences Biotechnology
Biochemistry, 69 (6): 1213-1216.
6. Arahal, D., R., Russell, H., Carol, V., Litchfield,
D., Melanie, R., Mormile, B., Tindall, J., Aharon, O., Victoria, B., Quesada,
E. and Ventosa, A. (2007). Recommended minimal standards for describing new
taxa of the family Halomonadaceae. Systematic Evolutionary Microbiology,
57: 2436–2446.
7. Isnansetyo, A. and Kamei, Y. (2003).
MC21-A, a bactericidal antibiotic produced by a new marine bacterium, Pseudoalteromonas
phenolic sp. Nov. O-BC30T, against methicillin-resistant Staphylococcus
aureus. Antimicrobial and Chemotheraphy, 47 (2): 480-488.
8. Bitzer, J., Große, T., Wang, L., Lang, S.,
Beil, W. and Zeeck, A. (2006). New aminophenoxazinones from a marine Halomonas
sp.: fermentation, structure elucidation and biological activity. Antibiotics, 59 (2): 86-92.
9. Mabinya, L., V., Cosa, S., Mkwetshana, N. and
Okoh, A., I. (2011). Halomonas sp. OKOH – a marine bacterium isolated
from the bottom sediment of algoa bay – produces a polysaccharide
bioflocculant: partial characterization and biochemical analysis of its
properties. Molecules, 16: 4358-4370.
10. Ojima, T., Saburi, W., Yamamoto, T. and
Kudo, T. (2012). Characterization of Halomonas sp. strain H11
α-glucosidase activated by monovalent cations and its application for
efficient synthesis of α-D-glucosylglycerol. Applied and Environmental
Microbiology, 78 (6): 1836-1845.
11. Chen, W. and Liu, Y. (2013). Isolation and
identification of Halomonas sp. ZSCW-10: a moderately halophilic
bacteria strain with cellulose activity. Advances Materials Research, 749:
236-241.
12. Donio, M., B., S., Ronica, F., A., Viji,
V., T., Velmurugan, S., Jenifer, J., S., C., A., MichaelBabu, M., Dhar, P. and
Citarasu, T. (2013). Halomonas sp. BS4, a biosurfactant producing
halophilic bacterium isolated from solar salt works in India and their
biomedical importance. SpringerPlus, 2 (149): 1-10.
13. Juan, L., Fuchao, L., Ling, L., Peng, J. and
Zhaopu, L. (2013). Inhibitory activity of an extract from a marine bacterium Halomonas
sp. HSB07 against the red-tide microalga Gymnodinium sp. (Pyrropphyta). Chinese
Journal of Oceanology and Limnology, 31 (6): 1241-1247.
14. Peschel, A. and Sahl, H.S. (2006). The
co-evolution of host cationic antimicrobial peptides and microbial resistance. Nature
Reviews Microbiology, 4: 529-536.
15. Kanagasabhapathy, M. and Nagata, S. (2008).
Cross-species induction of antibacterial activity produced by epibiotic
bacteria isolated from Indian marine sponge Pseudoceratina purpurea. World
Microbiology Biotechnology, 24: 687–691.
16. Di Maro ,A., De Maio, A., Castellano, S.,
Parente, A., Farina, B., and Faraone-Mennella, M.R. (2009). The
ADP-ribosylating thermozyme from Sulfolobus solfataricus is a DING
protein. Biology Chemistry, S1431–S6730.
17. Ellen, A., F., Rohulya, O., V.,Fusetti, F.,
Wagner, M., Albers, S-V. and Arnold J. M. Driessen. (2011). The sulfolobicin
genes of Sulfolobus acidocaldarius encode novel antimicrobial proteins. Bacteriology,
193 (17): 4380-4387.
18. Pantazaki, A., A., Tsolkas, G., P. and
Kyriakidis, D.A. (2008). A DING phosphatase in Thermus thermophilus. Amino
Acids, 34: 437 – 448.
19. Berna, A., Bernier, F., Chabriere, E.,
Perera, T. and Scott, K. (2008). DING proteins; novel members of a prokaryotic
phosphate-binding protein superfamily which extends into the eukaryotic
kingdom. Biochemistry Cell Biology, 40: 170–175.
20. Collombet, J.,M., Elias, M., Gotthard, G.,
Four, E., Renault, F., Joffre, A., Baubichon, D., Rochu, D. and Chabričre, E.
(2010). Eukaryotic DING proteins are endogenous: an immunohistological study in
mouse tissues. PLoS ONE e9099, 5
(2): 1-7.
21. Longeon, A., Peduzzi, J., Barthe´, M. L.,
Corre, S., Nicolas, J-L. and Guyot, M. (2004). Purification and partial
identification of novel antimicrobial protein from marine bacterium Pseudoalteromonas species strain X153.
Marine Biotechnology, 6: 633–641.
22. Atirah., W.S.N. (2011). Identification and
characterization of antimicrobial peptides in Halomonas sp. from Lutjanus
erythropterus epidermis. Msc. Thesis. Universiti Teknologi Malaysia,
Skudai.
23. Azemin, A. and Omar, M., S., S. (2014).
Detection and activity of bacteriocin produced by Halomonas sp. strain
M3. Proceeding of International Postgraduate Conference on Science and
Mathematics, 2: 47-58.
24. Giek Far, C. and Rashid, N. A. (2005).
Flavin reductase from Citrobacter freundii A1. Ph.D. Thesis.
Universiti Teknologi Malaysia, Skudai.
25. Chong, K., Joshi, S., Jin, L., T. and
Shu-chien, A., C. (2005). Proteomics profiling of epidermal mucus secretion of
a cichlid (Symphysodon aequifasciata) demonstrating parental care
behaviour. Proteomics, 5: 2251-2258.
26. Mousavi, S. M., Wilson, G., Raftos, D., Mirzargar, S. S. and Omidbaigi,
R. (2011). Antibacterial activities of a new combination of essential oils
against marine bacteria. Aquaculture International, 19 (1): 205-214.
27. Bringans, S., Eriksen, S., Kendrick, T.,
Gopalakrishnakone, P., Livk, A., Lock, R. and Lipscombe, R. (2008). Proteomic
analysis of the venom of Heterometrus longimanus (Asian black
scorpion). Animal Proteomic, 8 (5): 1081-1096.
28. Lewis, A.,P. and Crowther, D. (2005). DING
proteins are from Pseudomonas. FEMS Microbiology Letters, 252:
215–222.
29. Scott, K. and Wu, L. (2005). Functional
properties of a recombinant bacterial DING protein: comparison with a
homologous human protein. Biochim Biophys Acta, 1744: 234–244.
30. Ahn, S., Moniot, S., Elias, M., Chabriere,
E. and Kim, D. (2007). Structure-function relationships in a bacterial DING
protein. FEBS Letters, 581: 3455–3460.
31. Tan, A.,S. and Worobec, E.,A. (1993).
Isolation and characterization of two immunochemically distinct alkaline
phosphatases from Pseudomonas aeruginosa. FEMS Microbiology Letters, 106:
281–286.
32. Ball, G., Durand, E., Lazdunski, A. and
Filloux, A. (2002). A novel type II secretion system in Pseudomonas
aeruginosa. Molecular Microbiology, 43: 475–485.
33. Berna, A., Scott, K., Chabriere, E. and Bernier, F.
(2009). The DING family of proteins: ubiquitous in eukaryotes, but where are
the genes?. Bioessays, 31: 570–580.
34. Perera, T., Berna, A., Scott, K., Lemaitre-Guillier, C. and Bernier, F.
(2008). Proteins related to St. John’s Wort p27SJ, a suppressor of HIV-1
expression, are ubiquitous in plants. Phytochemistry, 69: 865–872.
35. Zhang, X.,X., Scott, K., Meffin, R. and
Rainey, P.,B. (2007). Genetic characterization of psp encoding the DING protein
in Pseudomonas fluorescens SBW25. BMC Microbiology, 7: 114.
36. Belenky, M., Prasain, J., Kim, H. and
Barnes, S. (2003). DING, a genistein target in human breast cancer: a protein
without a gene. Nutrition, 133: 2497S–2501.
37. Riah, O., Dousset, J.,C., Bofill-Cardona,
E. and Courriere, P. (2000). Isolation and microsequencing of a novel cotinine
receptor. Cellular Molecule Neurobiology, 20: 653–664.