چکیده مقاله :
شناسایی باکتری های اسید لاکتیک تولید کننده باکتریوسین مورد توجه محققین بسیاری قرار گرفته است. بررسی ها نشان داده است که باکتریوسین های باکتری های اسید لاکتیک دارای خواص ضدباکتریایی قابلتوجهی در برابر باکتری های عامل مسمومیت می باشند و می توان این باکتریوسین ها را بهعنوان نگهدارنده های طبیعی مواد غذایی بهکار برد .در این پژوهش فعالیت ضدباکتریایی مایع رویی کشت 10 سویه از باکتریهای لاکتیکی استاندارد بومی (که قبلاً شیر خام و ماست محلی جداسازی شده بود) علیه ﻟﯿﺴﺘﺮﯾﺎ اﯾﻨﻮﮐﻮا، باسیلوس سرئوس، شیگلا فلکسنری، اشریشیا کولای، استافیلوکوکوس اورئوس، یرسینیا انتروکولیتیکا، کلبسیلا پنومونیه مورد ارزیابی قرار گرفت. سپس ماهیت پروتئینی عامل ضدباکتریایی با استفاده از آنزیم تریپسین بررسی گردید. هر آزمون در سه تکرار انجام و میانگین قطر هاله عدم رشد اندازه گیری سویهها با هم مقایسه شدند. طبق نتایج مطالعه، سویه های اسید لاکتیک توان ضدباکتریایی خوبی را در مقابل باکتریهای شاخص نشان دادند. همچنین سویه 15E با قطر هاله عدم رشد 03/6 میلیمتر علیه شیگلا فلکسنری و با قطر هاله عدم رشد 167/0 میلیمتر علیه یرسینیا انتروکولیتیکا بهترتیب بیشترین و کمترین بازدارندگی را نشان دادند. اشریشیا کولای و شیگلا فلکسنری حساس ترین و باسیلوس سرئوس مقاوم ترین سویه های شاخص در برابر باکتری های اسید لاکتیک بومی شناخته شدند. همچنین نتایج تیمار آنزیمی ماهیت پروتئینی عوامل آنتاگونیستی تولید شده توسط باکتریهای لاکتیکی را تأیید کرد. بر اساس یافتههای مطالعه، سویه بومی T2 را میتوان سویه ای با پتانسیل بسیار بالا برای مهار رشد باکتری های گرم منفی نظیر شیگلا فلکسنری، اشریشیا کولای، یرسینیا انتروکولیتیکا و کلبسیلا نومونیه در نظر گرفت.
چکیده انگلیسی:
Identification of lactic acid bacteria, bacteriocin producing has been noticeable by most researchers. The surveys showed that bacteriocins of lactic acid bacteria has antibacterial effect against bacteria of poisoning agent, so these bacteriocins could be used as natural preserving of food materials. In this research, antimicrobial activities of bacterial supernatant from 10 native strain of lactic acid bacteria existing in Biotechnology Research Institute of Iran (ABRII), Branch of North-West and West Region of Iran evaluated by disk diffusion assay against seven pathogen including Listeria innocua, Bacillus cereus, flexneri Shigella, Escherichia coli, Staphylococcus aureus, Yersinia entrocolitica, Klebsiella pneumonia. Each test had three replications and the inhibition zone diameter measured in strains and compared with each other. At next step, peptide identification of antibacterial agent was detected by trypsin enzyme treatment. The results represented that lactic acid strains represented good antimicrobial potential against seven bacteria of poisoning agent. Also 15E strain with 6.03mm and 0.167mm inhibition zone diameter versus flexneri shigella and Yersinia entrocolitica showed the highest and lowest rate of suppression, respectively. Escherichia coli and Shigella flexneri the sensitive and Bacillus cereus as resistant strain was detected against bacteriocins of lactic acid bacteria. Also enzyme treatment result confirm peptide identification of antibacterial agent. T2 native strain could consider as strain with high potential versus gram-negative bacteria like Flexneri shigella, E. coli, Yersinia entrocolitica, and Klebsiella pneumonia.
منابع و مأخذ:
Akhondzade, A., Razavi, V., Misaghi, A., AbbasiFar, R., Radmehr, B. and Khalighi, F. (2003). Effect of thyme essential oils on Salmonella typhimurium in brain and heart broth. Journal of Medicinal Plants, 8: 84-91.
● Alizadeh, B. and Tarinejad, A. (2010). Application of MSTATC software in statistical analysis: Setoodeh Publication. Tabriz. [In Persian]
● Campos, C.A., Rodríguez, Ó., Calo-Mata, P., Prado, M., and Barros-Velázquez, J. (2006). Preliminary characterization of bacteriocins from Lactococcus lactis, Enterococcus faecium and Enterococcusmundtii strains isolated from turbot (Psetta maxima). Food Research International. 39(3): 356-364.
● Chen, H. and Hoover, D. (2003). Bacteriocins and their food applications. Comprehensive Reviews in Food Science and Food Safety, 2(3): 82-100.
· Cleveland, J., Montville, T.J., Nes, I.F. and Chikindas, M.L. (2001). Bacteriocins: safe, natural antimicrobials for food preservation. International Journal of Food Microbiology. 71(1): 1-20.
· Dobson, A., Cotter, P.D., Ross, R.P. and Hill, C. (2012). Bacteriocin production: a probiotic trait? Applied and Environmental Microbiology, 78(1): 1-6.
· Fernandez, B., Le Lay, C., Jean, J. and Fliss, I. (2013). Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions. Journal of Applied Microbiology, 114(3): 877-885.
· Garneau, S., Martin, N.I. and Vederas, J.C. (2002). Two-peptide bacteriocins produced by lactic acid bacteria. Biochimie. 84(5): 577-592.
· Gholamzadeh, M.A., Hejazi, M.A. and Hosseinzadeh Gharaje, N. (2017). Determination of bacteriocin encoding gene in six native strains of Lactobacillus plantarum. Journal of Food Science and Technology, 66(14): 17-25. [In Persian]
· Harris, L., Daeschel, M., Stiles, M. and Klaenhammer, T. (1989). Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes. Journal of Food Protection, 52(6): 384-387.
· Hernandez, D., Cardell, E. and Zarate, V. (2005). Antimicrobial activity of lactic acid bacteria isolated from Tenerife cheese: initial characterization of plantaricin TF711, a bacteriocin‐like substance produced by Lactobacillus plantarum TF711. Journal of Applied Microbiology, 99(1): 77-84.
· Ivanova, I., Kabadjova, P., Pantev, A., Danova, S. and Dousset, X. (2000). Detection, purification and partial characterization of a novel bacteriocin substance produced by Lactococcus lactis subsp. lactis B14 isolated from boza-Bulgarian traditional cereal beverage. Biocatalysis, 41(6): 47-53.
· Liu, W., Zhang, L., Yi, H., Shi, J., Xue, C., Li, H., et al. (2014). Qualitative detection of class IIa bacteriocinogenic lactic acid bacteria from traditional Chinese fermented food using a YGNGV-motif-based assay. Journal of Microbiological Methods, 100: 121-127.
· Marie, K.P., François, Z.N., Abbasi, A., Anwar, F., Ali, S.A., Victor, S.D., et al. (2012). Characterization of a bacteriocin produced by Lactobacillus plantarum Lp6SH isolated from" Sha'a", a maize-based traditionally fermented beverage from Cameroon. International Journal of Biology, 4(2): 149-158.
· Miller, K., Ray, P., Steinmetz, T., Hanekamp, T. and Ray, B. (2005). Gene organization and sequences of pediocin AcH/PA‐1 production operons in Pediococcus and Lactobacillus plasmids. Letters in Applied Microbiology, 40(1): 56-62.
· Narimani, T., Tarinejad, A. and Hejazi, M.A. (2013). Isolation and identification of lactic acid bacteria from traditional dairy products of Kleibar, Heris and Varzaghan. Food Hygiene, 3(3): 23-37. [In Persian]
· Pandey, N., Malik, R., Kaushik, J. and Singroha, G. (2013). Gassericin A: a circular bacteriocin produced by Lactic acid bacteria Lactobacillus gasseri. World Journal of Microbiology and Biotechnology, 29(11): 1977-1987.
· Pingitore, E.V., Salvucci, E., Sesma, F. and Nader-Macias, M.E. (2007). Different strategies for purification of antimicrobial peptides from lactic acid bacteria (LAB). Communicating Current Research and Educational Topics and Trends in Applied Microbiology, 1: 557-568.
· Sifour, M., Tayeb, I., Haddar, H.O., Namous, H. and Aissaoui, S. (2012). Production and characterization of bacteriocin of Lactobacillus plantarum F12 with inhibitory activity against Listeria monocytogenes. Online Journal of Science and Technology. 2: 55-61.
· Sip, A., Więckowicz, M., Olejnik-Schmidt, A. and Grajek, W. (2012). Anti-Listeria activity of lactic acid bacteria isolated from golka, a regional cheese produced in Poland. Food Control, 26(1): 117-124.
· Tafreshi, S.Y.H., Mirdamadi, S., Norouzian, D., Khatami, S. and Sardari, S. (2010). Effect of non-nutritional factors on nisin production. African Journal of Biotechnology, 9: 26-34.
· Van Reenen, C., Chikindas, M., Van Zyl, W. and Dicks, L. (2003). Characterization and heterologous expression of a class IIa bacteriocin, plantaricin 423 from Lactobacillus plantarum 423, in Saccharomyces cerevisiae. International Journal of Food Microbiology, 81(1): 29-40.
_||_Akhondzade, A., Razavi, V., Misaghi, A., AbbasiFar, R., Radmehr, B. and Khalighi, F. (2003). Effect of thyme essential oils on Salmonella typhimurium in brain and heart broth. Journal of Medicinal Plants, 8: 84-91.
● Alizadeh, B. and Tarinejad, A. (2010). Application of MSTATC software in statistical analysis: Setoodeh Publication. Tabriz. [In Persian]
● Campos, C.A., Rodríguez, Ó., Calo-Mata, P., Prado, M., and Barros-Velázquez, J. (2006). Preliminary characterization of bacteriocins from Lactococcus lactis, Enterococcus faecium and Enterococcusmundtii strains isolated from turbot (Psetta maxima). Food Research International. 39(3): 356-364.
● Chen, H. and Hoover, D. (2003). Bacteriocins and their food applications. Comprehensive Reviews in Food Science and Food Safety, 2(3): 82-100.
· Cleveland, J., Montville, T.J., Nes, I.F. and Chikindas, M.L. (2001). Bacteriocins: safe, natural antimicrobials for food preservation. International Journal of Food Microbiology. 71(1): 1-20.
· Dobson, A., Cotter, P.D., Ross, R.P. and Hill, C. (2012). Bacteriocin production: a probiotic trait? Applied and Environmental Microbiology, 78(1): 1-6.
· Fernandez, B., Le Lay, C., Jean, J. and Fliss, I. (2013). Growth, acid production and bacteriocin production by probiotic candidates under simulated colonic conditions. Journal of Applied Microbiology, 114(3): 877-885.
· Garneau, S., Martin, N.I. and Vederas, J.C. (2002). Two-peptide bacteriocins produced by lactic acid bacteria. Biochimie. 84(5): 577-592.
· Gholamzadeh, M.A., Hejazi, M.A. and Hosseinzadeh Gharaje, N. (2017). Determination of bacteriocin encoding gene in six native strains of Lactobacillus plantarum. Journal of Food Science and Technology, 66(14): 17-25. [In Persian]
· Harris, L., Daeschel, M., Stiles, M. and Klaenhammer, T. (1989). Antimicrobial activity of lactic acid bacteria against Listeria monocytogenes. Journal of Food Protection, 52(6): 384-387.
· Hernandez, D., Cardell, E. and Zarate, V. (2005). Antimicrobial activity of lactic acid bacteria isolated from Tenerife cheese: initial characterization of plantaricin TF711, a bacteriocin‐like substance produced by Lactobacillus plantarum TF711. Journal of Applied Microbiology, 99(1): 77-84.
· Ivanova, I., Kabadjova, P., Pantev, A., Danova, S. and Dousset, X. (2000). Detection, purification and partial characterization of a novel bacteriocin substance produced by Lactococcus lactis subsp. lactis B14 isolated from boza-Bulgarian traditional cereal beverage. Biocatalysis, 41(6): 47-53.
· Liu, W., Zhang, L., Yi, H., Shi, J., Xue, C., Li, H., et al. (2014). Qualitative detection of class IIa bacteriocinogenic lactic acid bacteria from traditional Chinese fermented food using a YGNGV-motif-based assay. Journal of Microbiological Methods, 100: 121-127.
· Marie, K.P., François, Z.N., Abbasi, A., Anwar, F., Ali, S.A., Victor, S.D., et al. (2012). Characterization of a bacteriocin produced by Lactobacillus plantarum Lp6SH isolated from" Sha'a", a maize-based traditionally fermented beverage from Cameroon. International Journal of Biology, 4(2): 149-158.
· Miller, K., Ray, P., Steinmetz, T., Hanekamp, T. and Ray, B. (2005). Gene organization and sequences of pediocin AcH/PA‐1 production operons in Pediococcus and Lactobacillus plasmids. Letters in Applied Microbiology, 40(1): 56-62.
· Narimani, T., Tarinejad, A. and Hejazi, M.A. (2013). Isolation and identification of lactic acid bacteria from traditional dairy products of Kleibar, Heris and Varzaghan. Food Hygiene, 3(3): 23-37. [In Persian]
· Pandey, N., Malik, R., Kaushik, J. and Singroha, G. (2013). Gassericin A: a circular bacteriocin produced by Lactic acid bacteria Lactobacillus gasseri. World Journal of Microbiology and Biotechnology, 29(11): 1977-1987.
· Pingitore, E.V., Salvucci, E., Sesma, F. and Nader-Macias, M.E. (2007). Different strategies for purification of antimicrobial peptides from lactic acid bacteria (LAB). Communicating Current Research and Educational Topics and Trends in Applied Microbiology, 1: 557-568.
· Sifour, M., Tayeb, I., Haddar, H.O., Namous, H. and Aissaoui, S. (2012). Production and characterization of bacteriocin of Lactobacillus plantarum F12 with inhibitory activity against Listeria monocytogenes. Online Journal of Science and Technology. 2: 55-61.
· Sip, A., Więckowicz, M., Olejnik-Schmidt, A. and Grajek, W. (2012). Anti-Listeria activity of lactic acid bacteria isolated from golka, a regional cheese produced in Poland. Food Control, 26(1): 117-124.
· Tafreshi, S.Y.H., Mirdamadi, S., Norouzian, D., Khatami, S. and Sardari, S. (2010). Effect of non-nutritional factors on nisin production. African Journal of Biotechnology, 9: 26-34.
· Van Reenen, C., Chikindas, M., Van Zyl, W. and Dicks, L. (2003). Characterization and heterologous expression of a class IIa bacteriocin, plantaricin 423 from Lactobacillus plantarum 423, in Saccharomyces cerevisiae. International Journal of Food Microbiology, 81(1): 29-40.