PEMURNIAN DAN KARAKTERISASI PEDIOSIN YANG DIPRODUKSI OLEH Pediococcus pentosaceus 2A2
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Published
27 December 2019
Keywords:
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characterization, molasses, pediocin, Pediococcus pentosaceus 2A2, purification
Abstract
Pediocin production by Pediococcus pentosaceus 2A2 was done in 30% molasses based medium at 37°C for 24 hours. The culture was then centrifuged to separate the cells and yield cell-free supernatant (CFS). Purification of the CFS was carried out by stepwise ammonium sulfate precipitation to achieve a 90% concentrate, followed by dialysis using membrane with a molecular weight cut off 2.0 kDa resulting in pediocin crude extract (PCE). The PCE was purified by cation exchange chromatography using SP Se-phadex C-25 and eluted with sodium acetat buffer at pH of 5.5; 6.0; 6.5; and 7.0 successively resulting in 4 fractions (F1, F2, F3, F4). Each purification step resulted in the increase in antimicrobial activity against Listeria monocytogenes ATCC 7644. The PCE had specific antimicrobial activity up to 5 fold higher than the CFS. F4 fraction, which showed the largest inhibition zone among the other fractions, had a very high specific activity up to 435 fold higher than the CFS. The SDS-PAGE analysis suggested that the molecular weight of F4 was approximately 5.9 kDa. The minimum inhibitory concentration (MIC) of F4 to inhibit 1 log (90%) of L. monocytogenes ATCC 7644 was 143 ppm. The inhibitory phenomenon observed under scanning electron microscope (SEM) showed that L. monocytogenes’s cells exposed to F4 experience morphological changes such as cells shrinkage.
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Altuntas EG, Cosansu S, Ayhan K. 2010. Some growth parameters and antimicrobial activity of a bacteriocin-producing strain Pediococcus aci-dilactici 13. Int J Food Microbiol 141: 28-31. DOI: 10.1016/j.ijfoodmicro.2010.04.024.
Anastasiadou S, Papagianni M, Filiousis G, Ambro-siadis I, Koidis P. 2008. Rapid quantifiable assessment of nutritional parameters influen-cing pediocin production by Pediococcus acidi-lactici NRRL B5627. Bioresour Technol 99: 6646-6650. DOI: 10.1016/j.biortech.2007.11. 068.
Arief II, Jakaria, Suryati T, Wulandari Z, Andreas E. 2013. Isolation and characterization of plantari-cin produced by Lactobacillus plantarum strains (IIA-1A5, IIA-1B1, IIA-2B2). Media Peternakan 2013: 91-100. DOI: 10.5398/medpet.2013.36.2. 91.
Arief II, Budiman C, Jenie BSL, Andreas E, Yuneni A. 2015a. Plantaricin IIA-1A5 from Lactobacillus plantarum IIA-1A5 displays bactericidal activity against Staphylococcus aureus. Benef Micro-bes 6: 603-613. DOI: 10.3920/BM2014.0064.
Arief II, Jenie BSL, Astawan M, Fujiyama K, Witarto AB. 2015b. Identification and probiotic charac-teristics of lactic acid bacteria isolated from Indonesian local beef. Asian J Anim Sci 9: 25-36. DOI: 10.3923/ajas.2015.25.36.
Bhattacharya S, Das A. 2010. Study of physical and cultural parameters on the bacteriocins pro-duced by lactic acid bacteria isolated from tradi-tional Indian fermented foods. Am J Food Technol 5: 111-120. DOI: 10.3923/ajft.2010.11 1.120.
Devi SM, Halami PM. 2011. Detection and Charac-terization of Pediocin PA-1/AcH like Bacteriocin Producing Lactic Acid Bacteria. Curr Microbiol 63: 181-185. DOI: 10.1007/s00284-011-9963-8.
Devi SM, Ramaswamy AM, Halami PM. 2014. In situ production of pediocin PA-1 like bacteriocin by different genera of lactic acid bacteria in soy-milk fermentation and evaluation of sensory properties of the fermented soy curd. J Food Sci Technol 51: 3325-3332. DOI: 10.1007/s131 97-012-0870-1.
Elayaraja S, Neelamegam A, Packiyam M, Thanga-vel B. 2014. Production, purification and cha-racterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spec-trum. Asian Pac J Trop Biomed 4: 305-311. DOI: 10.12980/APJTB.4.2014C537.
Garsa AK, Kumariya R, Sood SK, Kumar A, Kapila S. 2014a. Bacteriocin production and different strategies for their recovery and purification Probiotics Antimicrob Proteins 6: 47-58. DOI: 10.1007/s12602-013-9153-z.
Garsa AK, Kumariya R, Kumar A, Lather P, Kapila S, Sood SK. 2014b. Industrial cheese whey uti-lization for enhanced production of purified pe-diocin PA-1. Food Sci Technol 59: 656-665. DOI: 10.1016/j.lwt.2014.07.008.
Hata T, Tanaka R, Ohmomo S. 2010. Isolation and characterization of plantaricin ASM1: A new bacteriocin produced by Lactobacillus planta-rum A-1. Int J Food Microbiol 137: 94-99. DOI: 10.1016/j.ijfoodmicro.2009.10.021.
Jodoin J, Hincke MT. 2018. Histone H5 is a potent antimicrobial agent and a template for novel an-timicrobial peptides. Sci Rep 8: 1-15. DOI: 10.1 038/s41598-018-20912-1.
Kashani HH, Nikzad H, Mobaseri S, Hoseini ES. 2012. Synergism effect of nisin peptide in redu-cing chemical preservatives in food industry. Life Sci J 9: 496-501.
Kaur G, Singh P, Malik RK, Bhardwaj A. 2012. Me-chanism of nisin, pediocin 34, and enterocin FH99 resistance in Listeria monocytogenes. Probiotics Antimicrob Proteins 4: 11-20. DOI: 10.1007/s12602-011-9085-4.
Khan HU. 2012. The role of Ion Exchange Chroma-tography in purification and characterization of molecules. https://www.intechopen.com/books/ ion-exchange-technologies/the-role-of-ion-ex change-chromatography-in-purification-and-cha racterization-of-molecules [02 Agustus 2018].
Kingcha Y, Tosukhowong A, Zendo T, Roytrakul S, Luxananil P, Chareonpornsook K, Valyasevi R, Sonomoto K, Visessanguan W. 2012. Antiliste-ria activity of Pediococcus pentosaceus BCC 3772 and application as starter culture for Nham, a traditional fermented pork sausage. Food Control 25: 190-196. DOI: 10.1016/j.food cont.2011.10.005.
Lowry OH, Rosebrough NJ, Farr AL, Randall R.J. 1951. Protein measurement with the Folin phe-nol reagent. J Biol Chem 193: 265-275.
Ly KCD, Gabelli SB. 2014. Salting out of proteins using ammonium sulfate precipitation. Methods Enzymol 541: 85-94. DOI: 10.1016/B978-0-12-420119-4.00007-0.
Mehla J, Sood SK. 2011a. Acidic pH enhanced acti-vity/yield of an YGNGV motif containing anti-microbial peptide isolated and purified from Pe-diococcus pentosaceus NCDC273, a dairy strain. Int J Probiotics Prebiotics 6: 81-88.
Mehla J, Sood SK. 2011b. Substantiation in Entero-coccus faecalis of dose-dependent resistance and cross-resistance to pore-forming antimicro-bial peptides by use of a polydiacetylene-based colorimetric assay. Appl Environ Microbiol 77: 786-793. DOI: 10.1128/AEM.01496-10.
Nissen-Meyer JN, Rogne P, Oppegard C, Haugen HS, Kristiansen PE. 2009. Structure-function relationships of the non-lanthionine-containing peptide (class II) bacteriocins produced by gram-positive bacteria. Curr Pharm Biotechnol 10: 19-37. DOI: 10.2174/1389201097870486 61.
Mulyani S, Jenie BSL, Kusumaningrum HD, Arief II. 2019. Characterisation of crude bacteriocin pro-duced by Pediococcus pentosaceus 2A2 in en-riched molasses medium. Int Food Res J 26: 187-192.
Papagianni M, Anastasiadou S. 2009. Pediocins: The bacteriocins of Pediococci. Sources, pro-duction, properties and applications. Microb Cell Fact 8: 1-16. DOI: 10.1186/1475-2859-8-3.
Pharmacia Biotech. 2016. Sephadex ion exchange media. http://www.chembio.uoguelph.ca/educ mat/chm357/sephadex.pdf [1 November 2016].
Rodriguez JM, Martinez M, Kok J. 2002. Pediocin PA-1, a wide-spectrum bacteriocin from lactic acid bacteria. Crit Rev Food Sci Nutr 42: 91-121. DOI: 10.1080/10408690290825475.
Simha BV, Sood SK, Kumariya R, Garsa AK. 2012. Simple and rapid purification of pediocin PA-1 from Pediococcus pentosaceous NCDC 273 suitable for industrial application. Microbiol Res 167: 544-549. DOI: 10.1016/j.micres.2012.01. 001.
Sood SK, Simha BV, Kumariya R, Garsa AK, Mehla J, Meena S, Lather P. 2013. Highly specific culture-independent detection of YGNGV motif-containing pediocin-producing strains. Probio-tics Antimicro 5: 37-42. DOI: 10.1007/s1260201 2-9114-y.
Tiwari SK, Srivastava S. 2008. Purification and cha-racterization of plantaricin LR14: a novel bacte-riocin produced by Lactobacillus plantarum LR/ 14. Appl Microbiol Biotechnol 79: 759-767. DOI: 10.1007/s00253-008-1482-6.
Todorov SD, Dicks LMT. 2009. Bacteriocin produc-tion by Pediococcus pentosaceus isolated from Marula (Scerocarya birrea). Int J Food Microbiol 132: 117-126. DOI: 10.1016/j.ijfoodmicro.2009. 04.010.
Todorov SD, Wachsman M, Tome E, Dousset X, Destro MT, Dicks LMT, Franco BDGM, Velho MV, Drider D. 2010. Characterisation of an anti-viral pediocin-like bacteriocin produced by Ente-rococcus faecium. Food Microbiol 27: 869-879. DOI: 10.1016/j.fm.2010.05.001.
Wang G, Feng G, Synder AB, Manns D, Churey JJ, Worobo RW. 2014. Bactericidal thurincin H causes unique morphological changes in Baci-llus cereus F4552 without affecting membrane permeability. FEMS Microbiol Lett 357: 69-76. DOI: 10.1111/1574-6968.12486.
Authors
TriwulandhariN. W., JenieB. S. L., LioeH. N., & AriefI. I. (2019). PEMURNIAN DAN KARAKTERISASI PEDIOSIN YANG DIPRODUKSI OLEH Pediococcus pentosaceus 2A2. Jurnal Teknologi Dan Industri Pangan, 30(2), 91-99. https://doi.org/10.6066/jtip.2019.30.2.91
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