Characterization of antibacterial activity produced by Bacillus spp. isolated from honey and bee-associated products against foodborne pathogens
##plugins.themes.bootstrap3.article.sidebar##
##plugins.themes.bootstrap3.article.main##
Abstract
Abstract. Magdalena S, Anggelia, Yogiara. 2020. Characterization of antibacterial activity produced by Bacillus spp. isolated from honey and bee-associated products against foodborne pathogens. Bioteknologi 17: 51-59. Four potential isolates (Bacillus velezensis Y12, Bacillus amyloliquefaciens Y21, Bacillus amyloliquefaciens Y23, and Bacillus velezensis Y33) isolated from honey, propolis, and bee pollen from West Java, Indonesia showed antifungal and enzymatic activities. This study aimed to assay antibacterial activity against foodborne pathogens, determine the optimum condition for antibacterial compounds production, observe the effect of thermal and pH treatment on the antibacterial compounds, and detect the presence of antibacterial peptide biosynthesis gene. In this research, bacterial isolates showed antibacterial activity against Bacillus cereus ATCC 14579, Salmonella enterica 51741, and Salmonella Typhimurium ATCC 14028. The growth condition for antibacterial production of isolate B. velezensis Y12 was at the range of 25-60°C and pH 7-9. Meanwhile, the other three isolates showed the same pattern, also at the range of 37-60 °C. Antibacterial compounds against B. cereus were found heat stable at 4-90°C and active over pH 3-9. Cultivation of the isolates in BHIB and TSB did not significantly increase the antibacterial activity against all pathogens as compared to an LB medium. On the contrary, the antibacterial activity against S. typhimurium of the isolates cultivated in optimized medium two (yeast extract 32.5 g/L, glucose 33.4 g/L, MnSO4 0.042 g/L, CaCl2 0.031 g/L, KH2PO4 0.5 g/L, K2HPO4 0.5 g/L, (NH4)2SO4 1.0 g/L, and MgSO4 4.0 g/L) was significantly increased. Furthermore, seven antibacterial peptide biosynthesis genes primers were amplified from the genomic DNA of isolates B. velezensis Y12 and B. velezensis Y33 by PCR analysis. These genes were fenD, srfAA, bacA, bmyB, ituC, ituD, and bmyD. Meanwhile, there was no presence of ituC gene from the other isolates. This result suggests that isolate B. velezensisY12 might be the most potential isolate for antibacterial compounds production.
##plugins.themes.bootstrap3.article.details##
Alfonso A, Piccolo SL, Conigliaro G, Ventorino V, Burruano S, Moschetti G. 2012. Antifungal peptides produced by Bacillus amyloliquefaciens AG1 active against grapevine fungal pathogens. Ann Microbiol. 62(4):1593-1599.
Arguelles-Arias A, Ongena M, Halimi B, Lara Y, Brans A, Joris B, Fickers P. 2009. Bacillus amyloliquefaciens GA1 as a source of potent antibiotics and other secondary metabolites for biocontrol of plant pathogens. Microb Cell Fact. 8(1): 63-75.
Bizani D, Brandelli A. 2002. Characterization of a bacteriocin produced by a newly isolated Bacillus sp. strain 8 A. J Appl Microbiol. 93(3):512-519.
Chen XH, Koumoutsi A, Scholz R, Schneider K, Vater J, Sussmuth R, Piel J, Borris R. 2009. Genome analysis of Bacillus amyloliquefaciens FZB42 reveals its potential for biocontrol of plant pathogens. J Biotechnol. 140(1-2):27-37.
CLSI. 2018. Performance Standards of Antimicrobial Susceptibility Testing. 28th Edition. CLSI Supplement M100. Wayne, PA: Clinical and Laboratory Standards Institute.
Coutte F, Lecouturier D, Yahia SA, Leclere V, Bechet M, Jacques P, Dhulster P. 2010. Production of surfactin and fengycin by Bacillus subtilis in a bubbleless membrane bioreactor. Appl Microbiol Biotechnol. 87(2):499-507.
Gong AD, Li HP, Yuan QS, Song XS, Yao W, He WJ, Zhang JB, Liao YC. 2015. Antagonistic mechanism of iturin A and plipastatin A from Bacillus amyloliquefaciens S76-3 from wheat spikes against Fusarium graminearum. Plos One. 10(2):e0116871.
Hsieh F, Lin T, Meng M, Kao S. 2008. Comparing methods for identifying Bacillus strains capable of producing the antifungal lipopeptide iturin A. Curr Microbiol. 56(1):1-5.
Iqbal S, Qasim M, Begum F, Rahman H, Sajid I. 2018. Screening, characterization, and optimization of antibacterial peptides, produced by Bacillus safensis strain MK-12 isolated from waste dump soil KP, Pakistan. BioRXIV. 308205.
Jin H, Li K, Niu Y, Guo M, Hu C, Chen S, Huang F. 2015. Continuous enhancement of iturin A production by Bacillus subtilis with a stepwise two-stage glucose feeding strategy. BMC Biotechnol. 15(1):53-62.
Kiranmayi MU, Sudhakar P, Sreenivasulu K, Vijayalakshmi M. 2011. Optimization of culturing conditions for improved production of bioactive metabolites by Pseudonocardia sp. VUK-10. Mycobiology. 39(3):174-81.
Korenblum E, von der Weld I, Santos ALS, Rosado AS, Sebastian GV, Coutinho CMLM, Magalhaes FCM, de Paiva MM, Seldin L. 2005. Production of antimicrobial substances by Bacillus subtilis LFE-1, B. firmus H2O-1 and B. licheniformis T6-5 isolated from an oil reservoir in Brazil. J Appl Microbiol. 98(3):667-675.
Kumar SN, Siji JV, Ramya R, Nambisan B, Mohandas C. 2012. Improvement of antimicrobial activity of compounds produced by Bacillus sp. associated with a Rhabditid sp. (entomopathogenic nematode) by changing carbon and nitrogen sources in fermentation media. J Microbiol Biotechnol Food Sci. 1(6):1424-1438.
Lee H, Churey JJ, Worobo RW. 2008. Antimicrobial activity of bacterial isolates from different floral sources of honey. Int J Food Microbiol. 126:240-244.
Lertcanawanichakul M, Sawangnop S. 2008. A comparison of two methods used for measuring the antagonistic activity of Bacillus species. Walailak J Sci Tech. 5(2):161-171.
Lisboa MP, Bonatto D, Bizani D, Henriques JAP, Brandelli A. 2006. Characterization of a bacteriocin-like substance produced by Bacillus amyloliquefaciens isolated from the Brazilian Atlantic forest. Int Microbiol. 9(2):111-118.
Li X, Zhang Y, Wei Z, Guan Z, Cai Y, Liao X. 2016. Antifungal activity of isolated Bacillus amyloliquefaciens SYBC H47 for the biocontrol of peach gummosis. Plos One. 11(9): e0162125.
Liu X, Ren B, Gao H, Liu M, Dai H, Song F, Yu Z, Wang S, Hu J, Kokare CR, Zhang L. 2012. Optimization for the production of surfactin with a new synergistic antifungal activity. Plos One. 7(5):e34430.
Maget-Dana R, Peypoux F. 1994. Iturins, a special class of pore-forming lipopeptides: biological and physicochemical properties. Toxicology. 87(3):151-74.
Martinez-Klimova E, Rodríguez-Peña K, Sánchez, S. 2017. Endophytes as sources of antibiotics. Biochem Pharmacol. 134:1-17.
Meena KR, Kanwar SS. 2015. Lipopeptides as the antifungal and antibacterial agents: applications in food safety and therapeutics. Biomed Res Int. 1(1):1-9.
Moshafi MH, Forootanfar H, Ameri A, Shakibaie M, Dehgan-Noudeh G, Razavi M. 2011. Antimicrobial activity of Bacillus sp. strain FAS1 isolated from soil. Pak J Pharm Sci. 24(3):269-275.
Mosquera S, Gonzalez-Jaramillo LM, Orduz S, Villegas-Escobar V. 2014. Multiple response optimization of Bacillus subtilis EA-CB0015 culture and identification of antifungal metabolites. Biocatal Agric Biotechnol. 3(4):378-385.
Mora I, Cabrefiga J, Montesinos E. 2011. Antimicrobial peptide genes in Bacillus strains from plant environments. Int Microbiol. 14(4):213-223.
Motta AS, Cladera-Olivera F, Brandelli A. 2004. Screening for animicrobial activity among bacteria isolated from the Amazon Basin. Braz J Microbiol. 35(4):307-310.
Muhammad N, Akbar A, Shah A, Abbas G, Hussain M, Khan TA. 2015. Isolation optimization and characterization of antimicrobial peptide producing bacteria from soil. J Anim Plant Sci. 25(4):1107-1113.
Palazzini JM, Dunlap CA, Bowman MJ, Chulze SN. 2016. Bacillus velezensis RC 218 as a biocontrol agent to reduce Fusarium head blight and deoxynivalenol accumulation: Genome sequencing and secondary metabolite cluster profiles. Microbiol Res. 192(1):30-36.
Purnawidjaja I. 2018. Isolation and detection of potential microbial activity from honey and other bee-associated products in West Java [thesis]. Atma Jaya Catholic University of Indonesia. [Indonesian]
Ramachandran R, Chalasani AG, Lai R, Roy U. 2014. A broad-spectrum antimicrobial activity of Bacillus subtilis RLID 12.1. Scientific World J. 968487.
Ramarathnam R, Bo S, Chen Y, Fernando WGD, Xuewen G, Kievit T. 2007. Molecular and biochemical detection of fengycin- and bacillomycin D-producing Bacillus spp., antagonistic to fungal pathogens of canola and wheat. Can J Microbiol. 53(7):901-911.
Ripa FA, Nikkon F, Zaman S, Khondkar P. 2009. Optimal conditions for antimicrobial metabolites production from a new Streptomyces sp. RUPA-08PR isolated from Bangladeshi soil. Mycobiology. 37(3):211-214.
Risoen PA, Ronning P, Hegna IK, Kolsto AB. 2004. Characterization of a broad range antimicrobial substance from B. cereus. J Appl Microbiol. 96(4):648-655.
Sabate DC, Carillo L, Audisio MC. 2009. Inhibition of Paenibacillus larvae and Ascosphaera apis by Bacillus subtilis isolated from honeybee gut and honey samples. Res Microbiol. 160(3):193-199.
Sabate DC, Audisio MC. Inhibitory activity of surfactin, produced by different Bacillus subtilis subsp. subtilis strains, against Listeria monocytogenes sensitive and bacteriocin-resistant strains. Microbiol Res. 168(3):125-129.
Song B, Rong YJ, Zhao MX, Chi ZM. 2013. Antifungal activity of the lipopeptides produced by Bacillus amyloliquefaciens anti-CA against Candida albicans isolated from clinic. Appl Microbiol Biotechnol. 97(16):141-150.
Sutyak KE, Wirawan RE, Aroutcheva AA, Chikindas ML. 2008. Isolation of the Bacillus subtilis antimicrobial peptide subtilosin from the dairy product?derived Bacillus amyloliquefaciens. J Appl Microbiol. 104(4):1067-1074.
Teixeira ML, Rosa AD, Brandelli A. 2013. Characterization of an antimicrobial peptide produced by Bacillus subtilis subsp. spizezinii showing inhibitory activity towards Haemophilus parasuis. Microbiol. 159(5):980-988.
Tsuge K, Akiyama T, Shoda M. 2001. Cloning, sequencing, and characterization of the iturin A operon. J Bacteriol. 183(21):6265-6273.
Walsh G. 2002. Proteins: biochemistry and biotechnology. John Wiley & Sons, US.
Zhao X, Zhou ZJ, Han Y, Wang ZZ, Fan J, Xiao HZ. 2013. Isolation and identification of antifungal peptides from Bacillus BH072, a novel bacterium isolated from honey. Microbiol Res. 168(9): 598-606.