The Biological Activity of Some Extracted Proteins from Bacillus spp. Isolated from Soil Against Pathogenic Bacteria

Authors

DOI:

https://doi.org/10.23851/mjs.v30i4.685

Keywords:

Soil samples, Bacillus spp., Proteins, Antibacterial, Antibiofilm, Cytotoxcity

Abstract

Forty soil samples were collected from different regions in Baghdad city. Among 52 isolate, only 38 isolates (73.1 %) belong to Bacillus spp. All bacterial isolates have been submitted to the cultural, microscopical, biochemical examination and VITEK 2 System, Bacillus cereus was predominantly found 21 (55.3%) followed by Bacillus subtilis 11 (28.9%), then Bacillus amyloliquefaciens was 6 (15.8%). Bacillus cereus (B1) and Bacillus subtilis (B16) isolates were with the highest level with the antibacterial activity when selected. Extracellular proteins were extracted and purified with ammonium sulfate (80%) saturation, then submitted to Sepharose-6B gel filtration column. SDS-polyacrylamide gel electrophoresis was used to determine the molecular weights of proteins, and one band appeared to each of Bacillus cereus and Bacillus subtilis with molecular weight 68 and 50 KDa respectively. The results revealed that higher effect of antibacterial activity on Gram positive bacteria, and moderate on Gram negative bacteria except P.aeruginosa with no effect. The antibiofilm effect of the extract found higher effective against S.pyogens, moderate effect against each of (S.aureus, E.coli, S. marcescens, K. spp) and no effect against P.aeruginosa. The results of cytotoxicity showed that the purified extract was a higher cytotoxic effect on breast cancer MCF7 cells at 1000µg/ml concentration, while the crude extract had a higher cytotoxic effect on breast cancer MCF7 cells at 100µg/ml concentration.

Author Biography

  • Neihaya Heikmat Zaki, Department of Biology, Mustansiriyah University
    Department of Biology, Assistant Professor

References

A. Amin, M. A. Khan, M. Ehsanullah, U. Haroon, S. M. F. Azam, A. Hameed, "Production of peptide antibiotics by Bacillus spp. GU 057 indigenously isolated from saline soil", Brazilian Journal of Microbiology, vol. 43, no.4, pp. 1340-1346, 2012. [CrossRef] [PubMed]

G.M. Agg, D.G.I. Kingston, and D.J. Newman, "Anticancer agents from natural products", Boca Raton: CRC Press,2005.

R. K. Ganguly, S. K. SujoyMidya and Chakraborty, "Antioxidant and Anticancer Roles of a Novel Strain of Bacillus anthracis Isolated from Vermicompost Prepared from Paper Mill Sludge". BioMed Research International,Vol. 2018, pp.7, 2018. [CrossRef] [PubMed]

C. Avşar, H. Koyuncu, and S. Ara, "Isolation and Molecular Characterization of Bacillus spp. Isolated from Soil for Production of Industrial Enzymes", Biological and Chemical Research, pp. 72-86, 2017.

H.S. Alnahdi, "Isolation and screening of extracellular proteases produced by new Isolated Bacillus spp.", Journal of Applied Pharmaceutical Science, Vol. 2, no. 9, pp. 071-074, 2012. [CrossRef]

G. Sharma, S. Dang, S. Gupta, R. Gabrani, "the mechanism of action of bacteriocin from Bacillus subtilis GAS101", Med Princ Pract, Vol.27, pp.186-192, 2018. [CrossRef] [PubMed]

P. Singh, R. Sharma, A. K. Shukla, and R. Singh, "Isolation of Bacillus spp. from Soil for Antimicrobial Production and Antibiotic Resistance", Adv Biotech. & Micro., vol.8, no.4, pp.1-5, 2018.

J. F. Macfaddin, "Biochemical Tests for Identification of Medical Bacteria", 3th ed. lippincott Williams and Wilkins, pp.912 ,2000.

T. Vishwanatha, N.J. Spoorthi, V. Reena, B.C. Divyashree, K.G. Siddalingeshwara, J. Karthic and K.M. Sudipta, "Screening of substrates for protease production from Bacillus licheniformis", International Journal of Engineering Science and Technology, Vol. 2, no. 11, pp.6550-6554, 2010.

S. Stepanovic, D. Vukovic, V. Hola, G. Bonaventura, S. Djukic, I. Cirkovic and F. Ruzicka, "Quantification of biofilm in microtiter plates: Overview of testing Conditions and practical recommendations for assessment of biofilm production by Staphylococci", APMIS, Vol.115, pp. 891-899, 2007. [CrossRef] [PubMed]

NH. Zaki, And Z. Husain, "Investigating the effect of Biosynthesized Silver Nanoparticles as antibiofilm on Bacterial Clinical Isolates", Microbial pathogenesis, vol.116, pp.200-208, 2018. [CrossRef] [PubMed]

E. M. E. Safey, U. M. Abdul-Raouf, "Production, purification and characterization of protease enzyme from bacillus subtilis", International Conferences for Development and the Environment in the Arab World, Assiut Univ., vol.14, pp.23-25,2004.

B. W. Lash, T. H. Mysliwiec, and H. Gourama, "Detection and partial characterization of a broad-range bacteriocin produced by Lactobacillus plantarum (ATCC 8014)", Food microb. , vol.22, pp.199-204, 2005. [CrossRef]

A. Sanghi, N. Garg, V.K. Gupta, A. Mittal, R.C. Kuhad, "One step purification and charactirization of cellolase-free xylanase produced by alkalophilic Bacillus subtilis ASH" Brazilian Journal of Microbiology. Vol.41, pp. 467-476, 2010. [CrossRef] [PubMed]

V.K. Roy, N.S. Kumar, and G. Gurusubramanian, "Proteins structure, properties and their separation by SDS polyacrylamide gel electrophoresis". Sci.Vis. vol.12, pp.178-181, 2012.

G.A Costa, F. P. Rossatto, A. W. Medeiros, et al. "Evaluation antibacterial and antibiofilm activity of the antimicrobial peptide P34 against Staphylococcus aureus and Enterococcus faecalis", Anais da Academia Brasileira de Ciências , vol.90, no.1, pp. 73-84, 2018. [CrossRef] [PubMed]

R. I. Freshney, "Culture of Animal Cells. Amanual of Basic Technique and Specialized Applications". 6th ed. John Wiley and Sons, Inc,USA, 2010. [CrossRef]

D. C. Darling, and S. J. Morgan, "Animal cells: Culture and media", John Wiley and sons, Chichester, pp.90-118, 1994.

V. Nemudzivhadi and P. Masoko, "In Vitro Assessment of Cytotoxicity, Antioxidant, and Anti-Inflammatory Activities of Ricinus communis (Euphorbiaceae) Leaf Extracts", Evidence-Based Complementary and Alternative Medicine.Vol. 2014, pp.8, 2014. [CrossRef] [PubMed]

L. A. Betancur- Galvis, G. E. Morales, J.E. Forero and J. Roldan, "Cytotoxic and Antiviral Activities of Colombian Medicinal Plant Extracts of the Euphorbia genus", Mem. Inst. Oswaldo. Cruz., vol. 94, pp.541-46, 2002. [CrossRef] [PubMed]

F. J. Gutiérrez Mañero, A. Probanza, B. Ramos, et. Al., "Ecology, Genetic Diversity and Screening Strategies of Plant Growth Promoting Rhizobacteria (PGPR)", Journal of Plant Nutrition, vol.26, pp. 1101-1115,2003.

K. Santong, W. Chunglok, M. Lertcanawanichakul and P. Bangrak, "Screening and Isolation of Bacillus sp.Producing Thermotolerant Protease from Raw Milk", J Sci & Tech, vol.5, no.2, pp.151-160, 2008.

R.C. Patil and B.L. Jadhav, "Isolation and Characterization of Protease Producing Bacillus Species from Soil of Dairy Industry", Int.J.Curr.Microbiol.App.Sci, vol.6, no.6, pp. 853-860, 2017. [CrossRef]

MC. Wiener and PS. Horanyi, "How hydrophobic molecules traverse the outer membranes of Gram-negative bacteria", proc Natl Acad Sci USA, vol.108, pp. 10929 - 10 930, 2011. [CrossRef] [PubMed]

K.G. Torkar and B. B. Matijaši, "Partial characterisation of bacteriocins produced by Bacillus cereus isolates from milk and milk products",Food Technol. Biotechnol., vol.41, no.2, pp.121-129, 2003.

R. C. Lawrence, T. F. Frejer and B. Reiter, "The production and characterization of lipase from a Micrococcus and a Pseudomonas", J. Gen. Microbial., vol.48, pp. 401-418, 1967. [CrossRef] [PubMed]

J. E. Amadi and D. O. Adeniyi, "Mycotoxin production by fungi isolated from stored grains. African Journal of Biotechnology", vol.8, no.7, pp.2021 -2002, 2009.

M. N. Hanina, M. Hairul Shahril, I. Ismatul Nurul Asyikin, et. al., "Extracellular protein secreted by Bacillus subtilis ATCC21332 in the presence of streptomycin sulfate", World Academy of Science, Engineering and Technology International Journal of Bioengineering and Life Sciences, vol.8, no.8, 2014.

I. N. A. Ismail, H. M. Noor, H. S. Muhamad, et al., "Protein Secreted by Bacillus subtilis ATCC 21332 in the Presence of Allium sativum. American Journal of Biological, Chemical and Pharmaceutical Sciences", vol. 1, pp. 35 - 41, 2013.

M. Padmapriya and B. C. Williams, "Purification and characterization of neutral protease enzyme from Bacillus Subtilis", J. Microbiol. Biotech. Res., vol. 2, no.4, pp.612-618, 2012.

S. J. Todd, J. G. Arthur, A. J.G. Moir, et.al., "Genes of Bacillus cereus and Bacillus anthracis encoding proteins of the exosporium", Journal of bacteriology, pp. 3373-3378, 2003. [CrossRef] [PubMed]

A. Ghosh, K. Chakrabarti, D. Chattopadhyay, "Degradation of raw feather by a novel high molecular weightextracellular protease from newly isolated Bacillus cereus DCUW", J Ind Microbiol Biotechnol, vol.35, pp.825-834, 2008. [CrossRef] [PubMed]

C. Matz, D. McDougald, A. M. Moreno, P. Y. Yung, et.al., "Biofilm formation and phenotypic variation enhance predation-driven persistence of Vibrio cholera", Proc.Natl.Acad.Sci., USA., vol.102, pp.16819-16824, 2005. [CrossRef] [PubMed]

C. Prudêncio, M. Vieira, D.S. Teresinha, et al., "Strategies for the use of bacteriocins in gram-negative bacteria: relevance in food microbiology", J Food Sci Technol, vol.52, pp. 5408-5417, 2015. [CrossRef] [PubMed]

K. L. Nawrocki, E. K. Crispell and S.M. Mcbride, "Antimicrobial peptide resistance mechanisms of Gram-positive bacteria", Antibiotics, vol.3, pp. 461-492, 2014. [CrossRef] [PubMed]

M. C. Pedrosa, B. B. Golner, B. R. Goldin, S. Barakat, G. E. Dallal, and R. M. Russell, "Survival of yogurt-containing organisms and Lactobacillus gasseri (ADH) and their effect on bacterial enzyme activity in the gastrointestinal tract of healthy and hypochlorhydric elderly subjects", Am. J. Clin. Nutr., vol.61, pp.353-359,1995. [CrossRef] [PubMed]

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Published

15-01-2020

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Section

Biological Science

How to Cite

[1]
H. A. Ibrahim and N. H. Zaki, “The Biological Activity of Some Extracted Proteins from Bacillus spp. Isolated from Soil Against Pathogenic Bacteria”, Al-Mustansiriyah Journal of Science, vol. 30, no. 4, pp. 29–38, Jan. 2020, doi: 10.23851/mjs.v30i4.685.

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