Extracellular Protein Secreted by Bacillus subtilis ATCC21332 in the Presence of Streptomycin Sulfate

Authors: Hanina M. N., Hairul Shahril M., Ismatul Nurul Asyikin I., Abdul Jalil A. K., Salina M. R., Maryam M. R., Rosfarizan M.

Abstract:

The extracellular proteins secreted by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was carried out to determine the effect of Streptomycin Sulfate in regulating extracellular proteins secreted by Bacillus subtilis ATCC21332. Results of Microdilution assay showed that the Minimum Inhibition Concentration (MIC) of Streptomycin Sulfate on B. subtilis ATCC21332 was 2.5 mg/ml. The bacteria cells were then exposed to Streptomycin Sulfate at concentration of 0.01 MIC before being further incubated for 48h to 72 h. The extracellular proteins secreted were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile revealed that three additional bands with approximate sizes of 30 kDa, 22 kDa and 23 kDa were appeared for the treated bacteria with Streptomycin Sulfate. Thus, B. subtilis ATCC21332 in stressful condition with the presence of Streptomycin Sulfate at low concentration could induce the extracellular proteins secretion.

Keywords: Bacillus subtilis ATCC21332, Streptomycin Sulfate, extracellular proteins.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1094132

References:

[1] G.K. Sims, "Nitrogen starvation promotes biodegradation of N-heterocyclic compounds in soil”, Soil Bio. & Biochem., vol. 38, pp. 2478-2480, 2006.
[2] S.M. Amanda, C-O. Florencia & B. Adriano, "Screening for antimicrobial activity among bacteria isolated from the Amazon Basin”, Braz. J. Microb., vol. 35, pp. 307-310, 2004.
[3] V.A. Shinde, V. A., S. M. More & T. A. Kadam, "Antimicrobial Activity Of Phospholipid Compound Produced by Alkaliphilic Bacillus subtilis Isolated from Lonar Lake”. Int. J. Inno. Bio-Sci., vol. 2, pp. 172-175, 2012.
[4] M.S.S. Ann, X. Jiru, E. M. John, S. B. Ian & A. M. David, "Environmental Stress and Antibiotic Resistance in Food-Related Pathogens”, Appl. & Env. Microb., vol. 73, pp. 211-217, 2007.
[5] K.J. Boor, "Bacterial Stress Responses: What Doesn’t Kill Them Can Make Them Stronger”, PLoS Bio., vol. 4, pp. 18-20, 2006.
[6] Y.R. Brenda, R. C. Franklin, B. P. Daniel, W. S. Daniel & L. L. Mark, "Effects of sub-minimum inhibitory concentration antibiotic levels and temperature on growth kinetics and outer membrane protein expression in Mannheimia haemolytica and Haemophilus somnus”, The Can. J. Vet. Res.,Vol. 69, pp.1-10, 2004.
[7] E-B. Goh, G. Yim, W. Tsui, J. McClure, M.G. Suret & J. Davies, "Transcriptional modulation of bacterial gene expression by subinhibitory concentrations of antibiotics”, Proc. Natl. Acad. Sci. USA, vol. 99(26), pp. 17025-17030, 2002.v
[8] A. Fajardo, A. & Martı´nez, "Antibiotics as signals that trigger specific bacterial responses”, Curr. Opin. Microb., vol. 11, pp. 161-167, 2008.
[9] M. Tanaka, T. Hasegawa, A. Okamoto, K. Torii. & M. Ohta, "Effect of antibiotics on group A Streptococcus exoprotein production analyzed by two-dimensional gel electrophoresis”, Antimicrob. Agents. Chemother., vol. 49(1), pp. 88-96, 2005.
[10] J.N. Eloff, "A sensitive and quick microplate method to determine the minimal inhibitory concentration of plants extract for bacteria”, Planta Med. vol. 6, pp. 711-713, 1998.
[11] S.C. Chang & L. J. Cseke, "Extraction and purification of proteins”, Handbook of Molecular and Cellular in Biology and Medicine, vol. 3, 2004, pp 48-49.
[12] D. Rifkind & G. L. Freeman, The Nobel Prize Winning Discoveries in Infectious Diseases. 2005, pp 47-50.
[13] M.M. Nakano & P. Zuber, "Anaerobic growth of a "strict aerobe” (Bacillus subtilis)”, Annu. Rev. Microb., vol. 52, pp. 165-190, 1998.
[14] M.N Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin Ismail, M.R. Salina, M.R. Maryam, A.K. Abdul Jalil & M. Rosfarizan, "Protein Produced by Bacillus subtilis ATCC21332 in the Presence of Cymbopogon flexuosus Essential oil”. Key Eng. Mat., vol. 594-595, pp. 370-377, 2014.
[15] J.G. Black, Microbiology: Principles and Explorations, 6. ed. USA: John Wiley & Sons, 2005.
[16] K. Omura, M. Hitosugi, X. Zhu, M. Ikeda, H. Maeda & S. Tokudome, "A Newly Derived Protein from Bacillus subtilis natto with Both Antithrombotic and Fibrinolytic Effects”, J. Pharm. Sci., vol. 99, pp. 247-251, 2005.
[17] L. Korsten, L. & N. Cook, "Optimizing Culturing Conditions for Bacillus Subtilis”, South African Avocado Growers Association Yearbook, vol. 19, 1996, pp. 54-58.
[18] G-H. Kwon, P. Jae-Yong, K. Jong-Sang, L. Jinkyu, P. Cheon-Seok, Y. K. Dae & H. K. Jeong, "Cloning and Expression of a bpr Gene Encoding Bacillopeptidase F from Bacillus amyloliquefaciens CH86-1”, J. Microb. & Biotech., vol. 21, pp. 515-518, 2011.
[19] X.C. Wu, W. Lee, L. Tran & S. L Wong, "Engineering a Bacillus subtilis expression-secretion system with a strain deficient in six extracellular proteases”, J. Bacteriol., vol. 173, pp. 4952-4958, 1991.