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Purification, characterization and antibacterial spectrum of a compound produced by Bacillus cereus MTCC 10072

Archives of Microbiology volume 201, pages 1195–1205 (2019)Cite this article

Abstract

Awareness of the consumer has increased the demand of safe and chemical-free foods, and consequently it has increased the demand of antibacterial bioactive compounds. In the present study, antibacterial compound produced by a local bacterial isolate NSD MTCC 10072, showing antagonistic activity against six human pathogens, was isolated, partially purified and characterized. Maximum production of antibacterial compound was observed between 51 and 60 h after seeding. The antibacterial activity of the compound was found to be thermostable up to 80 °C for 60 min and its efficacy was very good between pH 4 and 12. Minimum inhibitory concentration (25.84 µg/µl) of the antibacterial compound was observed against Streptococcus aureus NICM 2901. GC–MS analysis of the bacterium secreted chemical compound (C11H18N2O2) was used to identify the antimicrobial compound as Pyrrolo(1,2-a) pyrazine-1,4-dione, hexahydro-3-(2-methylpropyl). In Silico studies showed that the antimicrobial compound is non-toxic, non-irritating and followed Lipinski-type properties which suggested that the compound could be used as potential drug against different human pathogens.

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References

  • Ahmad V, Iqbal AN, Haseeb M, Khan MS (2014) Antimicrobial Potential of bacteriocin producing Lysinibacillus jx16856 against foodborne bacterial and fungal pathogen, isolated from fruit and vegetable waste. Anaerobe 27:87–95

    CAS  Article  Google Scholar 

  • Ananou S, Munoz A, Martinez-Bueno M, Gonzalez-Tello P, Galvez A, Maqueda M, Valdivia E (2010) Evaluation of an enterocin AS-48 enriched bioactive powder obtained by spray drying. Food Microbiol 27:58–63

    CAS  Article  Google Scholar 

  • Anastasiadou S, Papagianni M, Filiousis G, Ambrosiadis I, Pediocin Koidis (2008) SA-1, an antimicrobial peptide from Pediococcus acidilactici NRRL B5627: production conditions, purification and characterization. Bioresour Technol 99:5384–5390. https://doi.org/10.1016/j.biortech.2007.11.015

    CAS  Article  PubMed  Google Scholar 

  • Atrih A, Rekhif N, Moir AJG, Lebrihi A, Lefebvre G (2001) Mode of action, purification and amino acid sequence of plantaricin C19, an anti-Listeria bacteriocin produced by Lactobacillus plantarum C19. Int J Food Microbiol 68:93–109

    CAS  Article  Google Scholar 

  • Bhaskar N, Sudeepa ES, Rashmi HN, Selvi AT (2007) Partial purification and characterization of protease of Bacillus proteolyticus CFR3001 isolated from fish processing waste and its antibacterial activities. Bioresour Technol 98:2758–2764

    CAS  Article  Google Scholar 

  • Castellano P, Belfiore C, Fadda S, Vignolo G (2008) GA review of bacteriocinogenic lactic acid bacteria used as bioprotective cultures in fresh meat produced in Argentina. Meat Sci 79(3):483–499. https://doi.org/10.1016/j.meatsci.2007.10.009

    CAS  Article  PubMed  Google Scholar 

  • Cetinkaya Y, Falk P, Mayhall CG (2000) Vancomycin- resistant Enterococci. Clin Microbiol Rev 13:686–707. https://doi.org/10.1128/cmr.13.4.686-707.2000

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Chen CM, Sebranek JG, Dickson JS, Mendonca AF (2004) Combining pediocin with post-packaging irradiation for control of Listeria monocytogenes on frankfurters. J Food Protect 67:1866–1875. https://doi.org/10.4315/0362-028x-67.9.1866

    CAS  Article  Google Scholar 

  • Cheng F, Li W, Zhou Y, Shen J, Wu Z, Liu G, Lee PW, Tang Y (2012) admetSAR: a comprehensive source and free tool for assessment of chemical ADMET properties. J Chem Inf Model 52(11):3099–3105. https://doi.org/10.1021/ci300367a

    CAS  Article  PubMed  Google Scholar 

  • Contreras BGL, De Vuyst L, Devreese B, Busanyova K, Raymaeckers J, Bosman F, Sablon E, Vandamme EJ (1997) Isolation, purification and amino acid sequence of lactobin A, one of the two bacteriocins produced by Lactobacillus amylovorus LMG, P-13139. Appl Environ Microbiol 63(1):13–20

    CAS  PubMed  PubMed Central  Google Scholar 

  • Dawson RMC, Elliot DC, Elliot WH, Jones KM (1969) pH, buffers and physiological media. In: Dawson RMC, Elliot DC, Elliot WH, Jones KM (eds) Data for biochemical research. Research University Press, Oxford, pp 501–502

    Google Scholar 

  • de Arauz LJD, Jozalaa AF, Mazzolab PG, Pennaa TV (2009) Nisin biotechnological production and application: a review. Trends Food Sci Technol 20:146–154. https://doi.org/10.1016/j.tifs.2009.01.056

    CAS  Article  Google Scholar 

  • Deegan LH, Cotter PD, Hill C, Ross P (2006) Bacteriocins: biological tools for bio-preservation and shelf-life extension. Int Dairy J 16:1058–1071

    CAS  Article  Google Scholar 

  • Dominguez APM, Bizani D, Olivera FC, Brandelli A (2007) Cerein 8A production in soybean protein using response surface methodology. Biochem Eng J 35:238–243

    CAS  Article  Google Scholar 

  • Ferchichi M, Frere J, Mabrouk K, Manai M (2001) Lactocin MMFII a novel class II bacteriocin produced by Lactococcus lactis MMFII, isolated from Tunisian dairy product. FEMS Microbiol Lett 205(1):49–55

    CAS  Article  Google Scholar 

  • Gillor O, Etzion A, Riley MA (2008) The dual role of bacteriocin as anti- and probiotics. Appl Microbiol Biotechnol 81:591–606. https://doi.org/10.1007/s00253-008-1726-5

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198. https://doi.org/10.1016/0003-9861(68)90654-1

    CAS  Article  PubMed  Google Scholar 

  • Hout E, Maghrous J, Barena-Gonzalez C (1996) Bacteriocin J46, a new bacteriocin produced by Lactococcus lactis subsp. cremoris J46: isolation and characterisation of the protein and its gene. Anaerobe 2(1):137–145. https://doi.org/10.1006/anae.1996.0018

    Article  Google Scholar 

  • Joana Gil-Chávez G, Villa JA, Ayala-Zavala JF, Basilio Heredia J, Sepulveda D, Yahia EM, González-Aguilar GA (2013) Technologies for extraction and production of bioactive compounds to be used as nutraceuticals and food ingredients: an overview. Compr Rev Food Sci Food Saf 12(1):5–23

    Article  Google Scholar 

  • Klaenhammer TR (1988) Bacteriocins of lactic acid bacteria. Biochimie 70:337–349

    CAS  Article  Google Scholar 

  • Kumar A, Saini S, Wray V, Nimtz M, Prakash A, John BN (2012) Characterisation of an antifungal compound produced by Bacillus sp. strain A5F that inhibits Sclerotinia sclerotiorum. J Basic Microbiol 52:1–19. https://doi.org/10.1002/jobm.201100463

    CAS  Article  Google Scholar 

  • Laemmili UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  Google Scholar 

  • Lee H, Churey J, Worobo R (2008) Purification and structural characterization of bacillomycin F produced by a bacterial honey isolate active against Byssochlamys fulva H25. J Appl Microbiol 105:663–673. https://doi.org/10.1111/j.1365-2672.2008.03797.x

    CAS  Article  PubMed  Google Scholar 

  • Lili H, Weiliang C, Yang L (2006) Production and partial characterization of bacteriocin like peptide from Bacillus licheniformis ZJU12. Microbiol Res 161(4):321–326

    Article  Google Scholar 

  • Lindgren SE, Dobrogosz WJ (1990) Antagonistic activities of lactic acid bacteria in food and feed fermentations. FEMS Microbiol Rev 87:149–164

    CAS  Article  Google Scholar 

  • Lipinski CA, Lombardo F, Dominy BW, Feeney PJ (1997) Experimental and computational approaches to estimate solubility and permeability in drug delivery and development settings. Adv Drug Deliv Rev 23:4–17. https://doi.org/10.1016/s0169-409x(96)00423-1

    Article  Google Scholar 

  • Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275. https://doi.org/10.1021/ci300367a

    CAS  Article  PubMed  Google Scholar 

  • Makroo RN, RainaV Bhatia A, Gupta R (2011) Evaluation of the red cell hemolysis in packed red cells during processing and storage. Asian J Transfus Sci 5(1):15–17

    CAS  Article  Google Scholar 

  • Millette M, Smoragiewicz W, Lacroix M (2004) Antimicrobial potential of immobilized Lactococcus lactis subsp. lactis ATCC 11454 against selected bacteria. J Food Prot 67(6):1184–1189

    CAS  Article  Google Scholar 

  • Motta AS, Lorenzini DM, Brandelli A (2007) Purification and partial characterization of an antimicrobial peptide produced by a novel Bacillus sp. isolated from the Amazon basin. Curr Microbiol 54:282–286. https://doi.org/10.1007/s00284-006-0414-x

    CAS  Article  PubMed  Google Scholar 

  • Oscariz JC, Lasa I, Pisabarro AG (1999) Detection and characterization of cerein 7, a newly bacteriocin produced by Bacillus cereus with a broad spectrum of activity. FEMS Microbiol Lett 178:337–341. https://doi.org/10.1111/j.1574-6968.1999.tb08696.x

    CAS  Article  PubMed  Google Scholar 

  • Pecci Y, Rivardo F, Martinotti MG, Allegrone G (2010) LC/ESI-MS/MS characterisation of lipopeptide biosurfactants produced by the Bacillus licheniformis V9T14 strain. J Mass Spectrom 45(7):772–778

    CAS  Article  Google Scholar 

  • Penna TCV, Jozala AF, Gentille TR, Pessoa JA, Cholewa O (2006) Detection of nisin expression by Lactococcus lactis using two susceptible bacteria to associate the effects of nisin with EDTA. Appl Biochem Biotechnol 121(124):334–346

    Article  Google Scholar 

  • Prema P, Smila D, Palavesam A, Immanuel G (2010) Production and characterization of an antifungal compound (3-phenyllactic acid) produced by Lactobacillus plantarum strain. Food Bioprocess Tech 3:379–386

    CAS  Article  Google Scholar 

  • Pyoung II, Kim JK, Sung SC, Joo H, Kim E, Yamaguchi T, Park D, Kim BG (2010) Characterization and structure identification of an antimicrobial peptide, hominicin, produced by Staphylococcus hominis MBBL 2-9. Biochem Biophys Res Commun 399:133–138

    Article  Google Scholar 

  • Raaijmakers JM, De Bruijn I, Nybroe O, Ongena M (2010) Natural functions of lipopeptides from Bacillus and Pseudomonas: more than surfactants and antibiotic. FEMS Microbiol Rev 34:1037–1062

    CAS  Article  Google Scholar 

  • Rayman K, Malik N, Hurst A (1983) Failure of nisin to inhibit outgrowth of Clostridium botulinum in model cured meat system. Appl Environ Microbiol 46:1450–1452

    CAS  PubMed  PubMed Central  Google Scholar 

  • Singh N, Rai V, Tripathi CKM (2013) Purification and chemical characterization of antimicrobial compounds from a new soil isolate Streptomyces rimosus MTCC 10792. Appl Biochem Microbiol 49(5):473–480

    CAS  Article  Google Scholar 

  • Teixeira ML, Cladera-Olivera F, Santosand JD, Brandelli A (2009) Purification and characterization of a peptide from Bacillus licheniformis showing dual antimicrobial and emulsifying activities. Food Res Int 42:63–68

    CAS  Article  Google Scholar 

  • Tichaczek PS, Nissen-Meyer J, Nes IF, Vogel RF, Hammes W (1992) Characterization of bacteriocins curvacin A from Lactobacillus curvatus LTH1174 and sakacin P from L. sake LTH673. Syst Appl Microbiol 15:460–468

    CAS  Article  Google Scholar 

  • Todorov SD, Dicks LMT (2005) Effect of growth medium on bacteriocin production by Lactobacillus plantarum ST194BZ, a strain isolated from Boza. Food Technol Biotechnol 43(2):165–173

    CAS  Google Scholar 

  • Todorov SD, Rachman C, Fourrier A, Dicks LMT, Van Reenen CA, Prevost H, Dousset X (2010) Characterization of bacteriocin produced by Lactobacillus sakei R1333 isolated from smoked salmon. Anaerobe 17(1):23–31

    Article  Google Scholar 

  • Van Reenan CA, Chikindas ML, Van-Zyl WH, Dicks LMT (2003) Characterization and heterologous expression of a class IIa bacteriocin, plantaricin 423 from Lactobacillus plantarum 423, in Saccharomyces cerevisiae. Int J Food Microbiol 81:29–40

    Article  Google Scholar 

  • Zhang B, Dong C, Shang Q, Cong Y, Kong W, Li P (2013) Purification and characterization of bacillomycin L produced by Bacillus amyloliquefaciens K103 from lemon. Appl Biochem Biotechnol 171:2262–2272

    CAS  Article  Google Scholar 

  • Zhu Z, Zhang G, Luo Y, Ran W, Shen KS (2012) Production of lipopeptides by Bacillus amyloliquefaciens in solid state fermentation using soybean flour and rice straw as the substrate. Bioresour Technol 112:254–260. https://doi.org/10.1016/j.biortech.2012.02.057

    CAS  Article  PubMed  Google Scholar 

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Acknowledgements

Authors are grateful to the University Grant Commission (UGC SAP), Department of Science and Technology (A-2/1754-57 dated 01/06/2016) and Haryana State Council of Science and Technology (HSCST 1803 dated 20/04/2016; HSCST 72/01 dated 01/01/2017) for providing the financial assistance under the MRP scheme, infrastructure facilities and fellowship, respectively.

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Affiliations

  1. Microbial Biotechnology laboratory, Department of Bio and Nano Technology, Guru Jambheshwar University of Science and Technology, Hisar, Haryana, 125001, India

    Narender kumar, Namita Singh, Rajneesh Jaryal, Chetna Bhandari, Jyoti Singh & Pallavi Thakur

  2. Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, 110054, India

    Pallavi Thakur

  3. Department of Agronomy, CCS Haryana Agricultural University, Hisar, Haryana, 125001, India

    Anil Duhan

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  1. Narender kumar
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  2. Namita Singh
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  3. Rajneesh Jaryal
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  4. Chetna Bhandari
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  5. Jyoti Singh
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  6. Pallavi Thakur
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Correspondence to Namita Singh.

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kumar, N., Singh, N., Jaryal, R. et al. Purification, characterization and antibacterial spectrum of a compound produced by Bacillus cereus MTCC 10072. Arch Microbiol 201, 1195–1205 (2019). https://doi.org/10.1007/s00203-019-01685-1

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  • DOI: https://doi.org/10.1007/s00203-019-01685-1

Keywords

  • Bacillus
  • Optimisation
  • Cytotoxicity
  • Antibacterial
  • Bio-control