شناسایی مولکولی و ارزیابی خواص ضد باکتریایی پدیوکوکوس پنتازاسئوس جدا شده از خمیرترش آرد کامل جو
محورهای موضوعی : میکروب شناسی غذاییعلیرضا صادقی 1 , مجتبی رئیسی 2 , مریم ابراهیمی 3 , بلال صادقی 4
1 - استادیار، گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان
2 - استادیار، مرکز تحقیقات سلامت غلات، دانشگاه علوم پزشکی گلستان، گرگان
3 - دانشجوی دکتری تخصصی، گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان
4 - استادیار، گروه بهداشت مواد غذایی دانشکده دامپزشکی، دانشگاه شهید باهنر کرمان، کرمان
کلید واژه: خواص ضد باکتریایی, خمیرترش آرد جو, پدیوکوکوس پنتازاسئوس, پالیده کشت,
چکیده مقاله :
سابقه و هدف: همواره شناسایی و تعیین ویژگی های جدایه های لاکتیکی زیست بوم هایی که کمتر مورد مطالعه قرار گرفته اند احتمال مواجهه با باکتری های دارای قابلیت های منحصر به فرد را در پی داشته است. این مطالعه با هدف شناسایی مولکولی و ارزیابی خواص ضد باکتریایی جدایه لاکتیکی غالب خمیرترش آرد جو انجام شد. مواد و روش ها: در این مطالعه تجربی پس از تهیه خمیرترش از آرد کامل جو، باکتری اسید لاکتیک غالب آن جداسازی گردید. در ادامه، جدایه لاکتیکی با توالی یابی محصولات PCR، شناسایی شد و ویژگی های ضد باکتریایی آن و پالیده کشت خام و خنثی شده فازهای رشد لگاریتمی و سکون جدایه یاد شده به ترتیب بر اساس روش های انتشار چاهک و میکرودایلوشن در برابر برخی از شاخص های باکتریایی غذازاد مورد ارزیابی قرار گرفت. یافته ها: توالی یابی محصولات PCR، موجب شناسایی پدیوکوکوس پنتازاسئوس به عنوان جدایه لاکتیکی غالب خمیرترش آرد جو شد. این جدایه از بین شاخص های باکتریایی مورد مطالعه، به شکل معنی داری (0.05>P) دارای اثر آنتاگونیستی بیشتری نسبت به لیستریا مونوسیتوژنز بود. علاوه بر این، پالیده خام حاصل از فاز رشد لگاریتمی جدایه یاد شده نسبت به سایر پالیده ها از فعالیت باکتریوسینی و همچنین تاثیر ضد باکتریایی بیشتری بر روی شاخص های باکتریایی غذازاد برخوردار بود. نتیجه گیری: پدیوکوکوس پنتازاسئوس جدا شده از خمیرترش آرد جو و پالیده های کشت آن دارای خاصیت بازدارندگی مناسبی در برابر شاخص های باکتریایی غذازاد مورد مطالعه بود. بنابراین می توان از این جدایه به عنوان کشت آغازگر و یا کشت همراه در فرآوری محصولات غذایی تخمیری به جای نگهدارنده های شیمیایی و آنتی بیوتیک های سنتزی با هدف بهبود ماندگاری و ارتقاء سلامت این فراورده ها استفاده نمود.
Background & Objectives: Usually identification and characterization of ecosystems isolates of lactic acid bacteria (LAB) which have been rarely studied lead to obtaining LAB with unique characteristics. The aims of this study were molecular characterization and evaluation of the antibacterial properties of dominant LAB isolated from whole barley sourdough. Materials & Methods: In this experimental study, first the sourdough was prepared from the whole barley flour, and subsequently its dominant LAB was isolated. LAB isolate was identified by sequencing of PCR products. Antibacterial properties of the isolate and its cell free culture filtrate (CCF) which was obtained from logarithmic and stationary phases as crud and naturalized form were also investigated for some food-borne indicator bacteria using well diffusion and microdilution methods, respectively. Results: Sequencing results of PCR products lead to identification of Pediococcus pentosaceus as the dominant LAB in whole barley sourdough. This LAB isolate had more antagonistic effect (p<0.05) on Listeria monocytogenes than other indicator bacteria. Furthermore, crud CCF obtained from logarithmic phase of the isolate had the highest bacteriocin activity and antibacterial ability in comparison to other CCFs. Conclusion: Whole barley sourdough P. pentosaceus isolate and its CCF have proper antibacterial properties against food-borne indicator bacteria used in this study. Therefore, P. pentosaceus has high potential to be used as microbial starter or adjunct culture in processing fermented foods instead of chemical preservatives or antibiotics in order to increase shelf life and safety of these products.
1. Hammes WP, Brandt MJ, Francis KL, Rosenheim J, Seitter MFH, Vogelmann SA. Microbial ecology of cereal fermentations. Trends Food Sci Tech. 2005; 16(1-3): 4-11.
2. Corsetti A, Settanni L. Lactobacilli in sourdough fermentation. Food Res Int. 2007; 40(5): 539-558.
3. Messens W, De Vuyst L. Inhibitory substances produced by Lactobacilli isolated from sourdoughs, a review. Int J Food Microbiol. 2002; 72(1): 31-43.
4. Simsek O, Hilmi Con A, Tulumoglu S. Isolating lactic starter cultures with antimicrobial activity for sourdough processes. Food Control. 2006; 17(4): 263-270.
5. Katina K, Arendt E, Liukkonen KH, Autio K, Flander L, Poutanen K. Potential of sourdough for healthier cereal products. Trends Food Sci Tech. 2005; 16(1-3): 104-112.
6. Poutanen K, Flander L, Katina K. Sourdough and cereal fermentation in a nutritional perspective. Food Microbiol. 2009; 26(7): 693-699.
7. Todorov S, Onno B, Sorokine O, Chobert JM, Ivanova I, Dousset X. Detection and characterization of a novel antibacterial substance produced by Lactobacillus plantarum ST31 isolated from sourdough. Int J Food Microbiol. 1999; 48(3): 167-177.
8. Katina K, Sauri M, Alakomi HL, Mattila-Sandholm T. Potential of lactic acid bacteria to inhibit rope spoilage in wheat sourdough bread. Food Sci Tech. 2002; 35(1): 38-45.
9. Pepe O, Blaiotta G, Moschetti G, Greco T, Villani F. Rope producing strains of Bacillus spp. from wheat bread and strategy for their control by lactic acid bacteria. Appl Environ Microb. 2003; 69(4): 2321-2329.
10. Leroy F, De Winter T, Adriany T, Neysens P, De Vuyst L. Sugars relevant for sourdough fermentation stimulate growth of and bacteriocin production by Lactobacillus amylovorus DCE 471. Int J Food Microbiol. 2006; 112(2): 102-111.
11. Mentes O, Ercan R, Akcelik M. Inhibitor activities of two Lactobacillus strains, isolated from sourdough, against rope-forming Bacillus strains. Food Control. 2007; 18(4): 359-363.
12. Corsetti A, Settanni L, Braga TM, Silva Lopes MF, Suzzi G. An investigation of the bacteriocinogenic potential of lactic acid bacteria associated with wheat (Triticum durum) kernels and non-conventional flours. Food Sci Tech. 2008; 41(7): 1173-1182.
13. Alizadeh S, Jamalifar H, Samadi N, Eaidi A, Fazeli M. Effect of sodium chloride on the kinetics of growth and antimicrobial potential of lactobacilli isolated from Iranian traditional sourdough. Iran J Nutr Sci Food Tech. 2010; 5(3): 47-56. [In Persian]
14. Zannini E, Garofalo C, Aquilanti L, Santarelli S, Silvestri G, Clementi F. Microbiological and technological characterization of sourdoughs destined for bread-making with barley flour. Food Microbiol. 2009; 26(7): 744-753.
15. Mariotti M, Garofalo C, Aquilanti L, Osimani A, Fongaro L, Tavoletti S, Hager AS, Clementi F. Barley flour exploitation in sourdough bread-making: a technological, nutritional and sensory evaluation. Food Sci Tech. 2014; 59(2): 973-980.
16. AACC International. Approved methods of the American association of cereal chemists. 11th Ed. The St. Paul. 2010.
17. Ferchichi M, Valcheva R, vost H, Onno B, Dousset X. Molecular identification of the microbiota of French sourdough using temporal temperature gradient gel electrophoresis. Food Microbiol. 2007; 24(7-8): 678-686.
18. Gulahmadov SG, Abdullaeva NF, Guseinova NF, Kuliev AA, Ivanova IV, Dalgalarondo M, Chobert JM, Haertlee T. Isolation and characterization of bacteriocin-like inhibitory substances from lactic acid bacteria isolated from Azerbaijan cheeses. Appl Biochem Microbiol. 2009; 45(3): 266-271.
19. Yang E, Fan L, Jiang Y, Doucette C, Fillmore S. Antimicrobial activity of bacteriocin producing lactic acid bacteria isolated from cheeses and yogurts. AMB Express. 2012; 2(1): 48-59.
20. Todorov SD, Dicks LMT. Pediocin ST18, an anti-listerial bacteriocin produced by Pediococcus pentosaceus ST18 isolated from boza, a traditional cereal beverage from Bulgaria. Process Biochem. 2005; 40(1): 365-370.
21. Ahmadova A, Todorov SD, Hadji-Sfaxi I, Choiset Y, Rabesona H, Messaoudi S, Kuliyev A, de Melo Franco BDG, Chobert JM, Haertlé T. Antimicrobial and antifungal activities of Lactobacillus curvatus strain isolated from homemade Azerbaijani cheese. Anaerobe. 2013; 20: 42-49.
22. Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Med Microbiol. 2009; 49(11): 1749-1755.
23. Chavan RS, Chavan SR. Sourdough technology, a traditional way for wholesome foods: a review. Comp Rev Food Sci Food Saf. 2011; 10(3): 170-183.
24. Sadeghi A, Mortazavi SA, Bahrami AR, Shahidi F, Matin MM, Khomeiri M. Evaluating the effect of plasmid on anti-ropiness activity of Lactobacillus plantarum isolated from Iranian traditional sourdough. Iran J Nutr Sci Food Tech. 2011; 6: 22 [In Persian].
25. Ammor S, Tauveron G, Dufour E, Chevallier I. Antibacterial activity of lactic acid bacteria against spoilage and pathogenic bacteria isolated from the same meat small-scale facility. Food Control. 2006; 17(6): 454-461.
26. Gálvez A, Abriouel H, López RL, Ben Omar N. Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol. 2007; 120(1-2): 51-70.
27. Corsetti A, Settanni L, Van Sinderen D. Characterization of bacteriocin-like inhibitory substances (BLIS) from sourdough lactic acid bacteria and evaluation of their in vitro and in situ activity. J Appl Microbiol. 2004; 96(3): 521-534.
28. Reis JA, Paula AT, Casarotti SN, Penna ALB. Lactic acid bacteria antimicrobial compounds: characteristics and applications. Food Eng Rev. 2012; 4(2): 124-140.
_||_1. Hammes WP, Brandt MJ, Francis KL, Rosenheim J, Seitter MFH, Vogelmann SA. Microbial ecology of cereal fermentations. Trends Food Sci Tech. 2005; 16(1-3): 4-11.
2. Corsetti A, Settanni L. Lactobacilli in sourdough fermentation. Food Res Int. 2007; 40(5): 539-558.
3. Messens W, De Vuyst L. Inhibitory substances produced by Lactobacilli isolated from sourdoughs, a review. Int J Food Microbiol. 2002; 72(1): 31-43.
4. Simsek O, Hilmi Con A, Tulumoglu S. Isolating lactic starter cultures with antimicrobial activity for sourdough processes. Food Control. 2006; 17(4): 263-270.
5. Katina K, Arendt E, Liukkonen KH, Autio K, Flander L, Poutanen K. Potential of sourdough for healthier cereal products. Trends Food Sci Tech. 2005; 16(1-3): 104-112.
6. Poutanen K, Flander L, Katina K. Sourdough and cereal fermentation in a nutritional perspective. Food Microbiol. 2009; 26(7): 693-699.
7. Todorov S, Onno B, Sorokine O, Chobert JM, Ivanova I, Dousset X. Detection and characterization of a novel antibacterial substance produced by Lactobacillus plantarum ST31 isolated from sourdough. Int J Food Microbiol. 1999; 48(3): 167-177.
8. Katina K, Sauri M, Alakomi HL, Mattila-Sandholm T. Potential of lactic acid bacteria to inhibit rope spoilage in wheat sourdough bread. Food Sci Tech. 2002; 35(1): 38-45.
9. Pepe O, Blaiotta G, Moschetti G, Greco T, Villani F. Rope producing strains of Bacillus spp. from wheat bread and strategy for their control by lactic acid bacteria. Appl Environ Microb. 2003; 69(4): 2321-2329.
10. Leroy F, De Winter T, Adriany T, Neysens P, De Vuyst L. Sugars relevant for sourdough fermentation stimulate growth of and bacteriocin production by Lactobacillus amylovorus DCE 471. Int J Food Microbiol. 2006; 112(2): 102-111.
11. Mentes O, Ercan R, Akcelik M. Inhibitor activities of two Lactobacillus strains, isolated from sourdough, against rope-forming Bacillus strains. Food Control. 2007; 18(4): 359-363.
12. Corsetti A, Settanni L, Braga TM, Silva Lopes MF, Suzzi G. An investigation of the bacteriocinogenic potential of lactic acid bacteria associated with wheat (Triticum durum) kernels and non-conventional flours. Food Sci Tech. 2008; 41(7): 1173-1182.
13. Alizadeh S, Jamalifar H, Samadi N, Eaidi A, Fazeli M. Effect of sodium chloride on the kinetics of growth and antimicrobial potential of lactobacilli isolated from Iranian traditional sourdough. Iran J Nutr Sci Food Tech. 2010; 5(3): 47-56. [In Persian]
14. Zannini E, Garofalo C, Aquilanti L, Santarelli S, Silvestri G, Clementi F. Microbiological and technological characterization of sourdoughs destined for bread-making with barley flour. Food Microbiol. 2009; 26(7): 744-753.
15. Mariotti M, Garofalo C, Aquilanti L, Osimani A, Fongaro L, Tavoletti S, Hager AS, Clementi F. Barley flour exploitation in sourdough bread-making: a technological, nutritional and sensory evaluation. Food Sci Tech. 2014; 59(2): 973-980.
16. AACC International. Approved methods of the American association of cereal chemists. 11th Ed. The St. Paul. 2010.
17. Ferchichi M, Valcheva R, vost H, Onno B, Dousset X. Molecular identification of the microbiota of French sourdough using temporal temperature gradient gel electrophoresis. Food Microbiol. 2007; 24(7-8): 678-686.
18. Gulahmadov SG, Abdullaeva NF, Guseinova NF, Kuliev AA, Ivanova IV, Dalgalarondo M, Chobert JM, Haertlee T. Isolation and characterization of bacteriocin-like inhibitory substances from lactic acid bacteria isolated from Azerbaijan cheeses. Appl Biochem Microbiol. 2009; 45(3): 266-271.
19. Yang E, Fan L, Jiang Y, Doucette C, Fillmore S. Antimicrobial activity of bacteriocin producing lactic acid bacteria isolated from cheeses and yogurts. AMB Express. 2012; 2(1): 48-59.
20. Todorov SD, Dicks LMT. Pediocin ST18, an anti-listerial bacteriocin produced by Pediococcus pentosaceus ST18 isolated from boza, a traditional cereal beverage from Bulgaria. Process Biochem. 2005; 40(1): 365-370.
21. Ahmadova A, Todorov SD, Hadji-Sfaxi I, Choiset Y, Rabesona H, Messaoudi S, Kuliyev A, de Melo Franco BDG, Chobert JM, Haertlé T. Antimicrobial and antifungal activities of Lactobacillus curvatus strain isolated from homemade Azerbaijani cheese. Anaerobe. 2013; 20: 42-49.
22. Jorgensen JH, Ferraro MJ. Antimicrobial susceptibility testing: a review of general principles and contemporary practices. Med Microbiol. 2009; 49(11): 1749-1755.
23. Chavan RS, Chavan SR. Sourdough technology, a traditional way for wholesome foods: a review. Comp Rev Food Sci Food Saf. 2011; 10(3): 170-183.
24. Sadeghi A, Mortazavi SA, Bahrami AR, Shahidi F, Matin MM, Khomeiri M. Evaluating the effect of plasmid on anti-ropiness activity of Lactobacillus plantarum isolated from Iranian traditional sourdough. Iran J Nutr Sci Food Tech. 2011; 6: 22 [In Persian].
25. Ammor S, Tauveron G, Dufour E, Chevallier I. Antibacterial activity of lactic acid bacteria against spoilage and pathogenic bacteria isolated from the same meat small-scale facility. Food Control. 2006; 17(6): 454-461.
26. Gálvez A, Abriouel H, López RL, Ben Omar N. Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol. 2007; 120(1-2): 51-70.
27. Corsetti A, Settanni L, Van Sinderen D. Characterization of bacteriocin-like inhibitory substances (BLIS) from sourdough lactic acid bacteria and evaluation of their in vitro and in situ activity. J Appl Microbiol. 2004; 96(3): 521-534.
28. Reis JA, Paula AT, Casarotti SN, Penna ALB. Lactic acid bacteria antimicrobial compounds: characteristics and applications. Food Eng Rev. 2012; 4(2): 124-140.
