جداسازی، شناسایی مولکولی و ارزیابی ویژگی های پروبیوتیکی لاکتوباسیلوس های غالب در خمیرترش آرد کامل گندم
محورهای موضوعی : میکروب شناسی غذایی
علیرضا صادقی
1
(استادیار، گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان)
مریم ابراهیمی
2
(دانشجوی دکتری تخصصی، گروه علوم و صنایع غذایی، دانشگاه علوم کشاورزی و منابع طبیعی گرگان، گرگان)
کلید واژه: پروبیوتیک, لاکتوباسیلوس, خمیرترش,
چکیده مقاله :
سابقه و هدف: استفاده روزافزون از فرآورده‌های غذایی صنعتی به جای محصولات سنتی، موجب افزایش حذف باکتری‌های پروبیوتیک می شود. این مطالعه با هدف جداسازی، شناسایی مولکولی و ارزیابی ویژگی های پروبیوتیکی لاکتوباسیلوس های غالب در خمیرترش آرد کامل گندم انجام شد. مواد و روش ها: در این مطالعــه تجربی پس از جداسازی و شناسایی لاکتوباسیلوس های غالب خمیرترش آرد کامل گندم، ویژگی های پروبیوتیکی آنها شامل زنده مانی در شرایط شبیه سازی شده دستگاه گوارش، اثرات ضد میکروبی در برابر استافیلوکوکوس اورئوس، لیستریا منوسیتوژنز، اشریشیا کلی، قابلیت تجمعی بر ضد اشریشیا کلی به عنوان یک عامل عفونی روده و همچنین مقاومت این جدایه ها در برابر برخی از آنتی بیوتیک های رایج مورد بررسی قرار گرفت. یافته ها: توالی یابی محصولات واکنش زنجیره ای پلی مراز، منجر به شناسایی لاکتوباسیلوس های اسیدوفیلوس، پلانتاروم، ساکی و برویس به عنوان لاکتوباسیلوس های غالب خمیرترش آرد کامل گندم شد. از میان این چهار جدایه، لاکتوباسیلوس پلانتاروم دارای بیشترین زنده مانی در شرایط شبیه سازی شده دستگاه گوارش بود. همچنین قطر هاله عدم رشد هر سه باکتری بیماری زا در مجاورت لاکتوباسیلوس پلانتاروم و لاکتوباسیلوس برویس به شکل معنی داری از دو جدایه لاکتیکی دیگر بیشتر بود. مقایسه قابلیت تجمعی جدایه های لاکتیکی بر ضد اشریشیا کلی نیز نشان داد که لاکتوباسیلوس برویس و لاکتوباسیلوس پلانتاروم از توانمندی بیشتری در این خصوص برخوردار می باشند. همچنین این دو جدایه به همراه لاکتوباسیلوس اسیدوفیلوس در برابر آنتی بیوتیک های استرپتومایسین، کانامایسین، ونکومایسین و نالیدیکسیک اسید مقاومت نشان دادند. نتیجه گیری: لاکتوباسیلوس برویس و لاکتوباسیلوس پلانتاروم جدا شده از خمیرترش آرد کامل گندم از قابلیت بالایی برای استفاده به عنوان باکتری های پروبیوتیک در مصارف مختلف غذایی و دارویی برخوردار می باشند.
Background & Objectives: Highly demands of consumption of industrial food products has led to removal of probiotic bacteria from daily diet. The aim of this study was evaluating the probiotic properties of dominant Lactobacillus isolated from whole wheat sourdough. Materials & Methods: In this experimental study, after isolation of dominant Lactobacillus from whole wheat sourdough, the isolates were identified by polymerase chain reaction (PCR). Probiotic properties of these isolates, including survival in simulated conditions of gastrointestinal tract, antimicrobial effects against prototype bacteria (Staphylococcus aureus, Listeria monocytogenes and Escherichia coli), the ability of co-aggregation with E. coli as an infection agent of intestine and resistance of these isolates against some of routine antibiotics were also investigated in this study. Results: Sequencing of the PCR products led to identification of Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus sakei and Lactobacillus brevis as dominant Lactobacillus in whole wheat sourdough. Among the mentioned isolates, Lactobacillus plantarum showed the maximum survival in simulated conditions of gastrointestinal tract. Inhibition zone diameter of three pathogenic bacteria in the presence of Lactobacillus plantarum and Lactobacillus brevis were significantly more than the others. Comparison between co-aggregation ability of isolated Lactobacillus with E. coli showed that Lactobacillus brevis and Lactobacillus plantarum were more effective. Furthermore, these strains along with L. acidophilus were resistant to Streptomycin, Kanamycin, Vancomycin and Nalidixic acid.Conclusion: Lactobacillus brevis and Lactobacillus plantarum isolated from whole wheat sourdough are highly useful to be uses as probiotic bacteria in food and medicinal applications.
1. Mendez-Vilas A. Microbial pathogens and strategies for combating them: science, technology and education. 1st ed. Zurbaran. Formatex Research Center; 2013.
2. Saarela M, Mogensen G, Fondén R, Mättö J, Mattila-Sandholm T. Probiotic bacteria: safety, functional and technological properties. J Biotechnol. 2000; 84(3): 197-215.
3. Poutanen K, Flander L, Katina K. Sourdough and cereal fermentation in a nutritional perspective. Food Microbiol. 2009; 26(7): 693-699.
4. 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.
5. Leroy F, De Vuyst L. Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci Tech. 2004; 15(2): 67-78.
6. Angmo K, Kumari A, Savitri A, Bhalla TC. Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of ladakh. Food Sci Tech. 2016; 66(1): 428-432.
7. Sabir F, Beyatli Y, Cumhur C. Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir. J Food Sci. 2010; 75(9): 568-573.
8. Masuda T, Kimura M, Okada S, Yasui H. Pediococcus pentosaceus Sn26 inhibits IgE production and the occurrence of ovalbumin-induced allergic diarrhoea in mice. Biosci Biotechnol Biochem. 2010; 74(2): 329-335.
9. Fernandez M, Boris S, Barbes C. Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract. J Appl Microbiol. 2003; 94(3): 449-455.
10. Hummel AS, Hertel C, Holzapfel WH, Franz C. Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. Appl Environ Microb. 2007; 73(3): 730-739.
11. Tabatabaei Yazdi F, Vasiee A, Alizadeh Behbahani B, Mortazavi SA. Evaluation of probiotic properties of lactic acid bacteria isolated from kimchi produced in Iran. Qom Univ Med Sci J. 2015; 9(5): 11-22. [In Persian]
12. Tofangsazan F, Shahidi F, Mortazavi SA, Milani E, Eshaghi Z. Investigation of antibacterial activity of lactic acid bacteria isolated from traditional Kordish cheese in comparison with commercial strains. Iran J Med Microbiol. 2013; 7(3): 34-41. [In Persian]
13. Narimani T, Tarinejad A. Isolation, biochemical and molecular identification of probiotic bacteria from traditional buffalo milk and yogurt of Khoi city. J Food Res. 2014; 24(3): 335-349. [In Persian]
14. Farahbakhsh M, Hakimi H, Bahram Abadi R, Zolfaghari M, Doraki N. Isolation of probiotic lactobacilli from traditional yogurts produced in rural Areas of Rafsanjan and their antimicrobial effects. J Rafsanjan Univ Med Sci. 2013; 12(9): 733-746. [In Persian]
15. 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.
16. Grzeskowiak L, Collado MC, Vesterlund S, Mazurkiewicz J, Salminen S. Adhesion abilities of commensal fish bacteria by use of mucus model system. Aquaculture 2011; 318(1-2): 33-36.
17. Zhang Y, Zhang L, Du M, Yi H, Guo C, Tuo Y, Han X, Li J, Zhang L, Yang L. Antimicrobial activity against Shigella sonnei and probiotic properties of wild lactobacilli from fermented food. Microbiol Res. 2011; 167(1): 27-31.
18. Nikoskelainen S, Ouwehand AC, Bylund G, Salminen S, Lilius EM. Immune enhancement in rainbow trout (Oncorhynchus mykiss) by potential probiotic bacteria (Lactobacillus rhamnosus). Fish Shellfish Immun. 2003; 15(5): 443-452.
19. Simsek O, Hilmi Con A, Tulumoglu S. Isolating lactic starter cultures with antimicrobial activity for sourdough processes. Food Control. 2006; 17(4): 263-270.
20. Rojo-Bezares B, Saenz Y, Poeta P, Zarazaga M, Ruiz-Larrea F, Torres C. Assessment of antibiotic susceptibility within lactic acid bacteria strains isolated from wine. Int J Food Microbiol. 2006; 111(3): 234-240
21. 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]
22. Khorasanchi N, Peighambardoust SH, Golshan Tafti A, Hejazi MA, Rafat SA. Evaluating the ability of liquid sourdough containing L. plantarum and L. reuteri starters in inhibition of bread mold spoilage. J Food Res. 2011; 21(3): 391-399. [In Persian]
23. Vinderola CG, Reinheimer JA. Lactic acid starter and probiotic bacteria: a comparative in vitro study of probiotic characteristics and biological barrier resistance. Food Res Int. 2003; 36(9): 895-904.
24. Schillinger U, Guigas C, Holzapfel WH. In vitro adherence and other properties of lactobacilli used in probiotic yoghurt-like products. Int Dairy J. 2005; 15(2): 1289-1297.
25. Collado MC, Surono I, Meriluoto J, Salminen S. Indigenous dadih lactic acid bacteria: cell-surface properties and interactions with pathogens. J Food Sci. 2007; 72(3): 89-93.
26. Dzidic S, Suskovic J, Kos B. Antibiotic resistance mechanisms in bacteria: biochemical and genetic aspects. Food Technol Biotech. 2008; 46(1): 11-21.
_||_1. Mendez-Vilas A. Microbial pathogens and strategies for combating them: science, technology and education. 1st ed. Zurbaran. Formatex Research Center; 2013.
2. Saarela M, Mogensen G, Fondén R, Mättö J, Mattila-Sandholm T. Probiotic bacteria: safety, functional and technological properties. J Biotechnol. 2000; 84(3): 197-215.
3. Poutanen K, Flander L, Katina K. Sourdough and cereal fermentation in a nutritional perspective. Food Microbiol. 2009; 26(7): 693-699.
4. 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.
5. Leroy F, De Vuyst L. Lactic acid bacteria as functional starter cultures for the food fermentation industry. Trends Food Sci Tech. 2004; 15(2): 67-78.
6. Angmo K, Kumari A, Savitri A, Bhalla TC. Probiotic characterization of lactic acid bacteria isolated from fermented foods and beverage of ladakh. Food Sci Tech. 2016; 66(1): 428-432.
7. Sabir F, Beyatli Y, Cumhur C. Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir. J Food Sci. 2010; 75(9): 568-573.
8. Masuda T, Kimura M, Okada S, Yasui H. Pediococcus pentosaceus Sn26 inhibits IgE production and the occurrence of ovalbumin-induced allergic diarrhoea in mice. Biosci Biotechnol Biochem. 2010; 74(2): 329-335.
9. Fernandez M, Boris S, Barbes C. Probiotic properties of human lactobacilli strains to be used in the gastrointestinal tract. J Appl Microbiol. 2003; 94(3): 449-455.
10. Hummel AS, Hertel C, Holzapfel WH, Franz C. Antibiotic resistances of starter and probiotic strains of lactic acid bacteria. Appl Environ Microb. 2007; 73(3): 730-739.
11. Tabatabaei Yazdi F, Vasiee A, Alizadeh Behbahani B, Mortazavi SA. Evaluation of probiotic properties of lactic acid bacteria isolated from kimchi produced in Iran. Qom Univ Med Sci J. 2015; 9(5): 11-22. [In Persian]
12. Tofangsazan F, Shahidi F, Mortazavi SA, Milani E, Eshaghi Z. Investigation of antibacterial activity of lactic acid bacteria isolated from traditional Kordish cheese in comparison with commercial strains. Iran J Med Microbiol. 2013; 7(3): 34-41. [In Persian]
13. Narimani T, Tarinejad A. Isolation, biochemical and molecular identification of probiotic bacteria from traditional buffalo milk and yogurt of Khoi city. J Food Res. 2014; 24(3): 335-349. [In Persian]
14. Farahbakhsh M, Hakimi H, Bahram Abadi R, Zolfaghari M, Doraki N. Isolation of probiotic lactobacilli from traditional yogurts produced in rural Areas of Rafsanjan and their antimicrobial effects. J Rafsanjan Univ Med Sci. 2013; 12(9): 733-746. [In Persian]
15. 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.
16. Grzeskowiak L, Collado MC, Vesterlund S, Mazurkiewicz J, Salminen S. Adhesion abilities of commensal fish bacteria by use of mucus model system. Aquaculture 2011; 318(1-2): 33-36.
17. Zhang Y, Zhang L, Du M, Yi H, Guo C, Tuo Y, Han X, Li J, Zhang L, Yang L. Antimicrobial activity against Shigella sonnei and probiotic properties of wild lactobacilli from fermented food. Microbiol Res. 2011; 167(1): 27-31.
18. Nikoskelainen S, Ouwehand AC, Bylund G, Salminen S, Lilius EM. Immune enhancement in rainbow trout (Oncorhynchus mykiss) by potential probiotic bacteria (Lactobacillus rhamnosus). Fish Shellfish Immun. 2003; 15(5): 443-452.
19. Simsek O, Hilmi Con A, Tulumoglu S. Isolating lactic starter cultures with antimicrobial activity for sourdough processes. Food Control. 2006; 17(4): 263-270.
20. Rojo-Bezares B, Saenz Y, Poeta P, Zarazaga M, Ruiz-Larrea F, Torres C. Assessment of antibiotic susceptibility within lactic acid bacteria strains isolated from wine. Int J Food Microbiol. 2006; 111(3): 234-240
21. 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]
22. Khorasanchi N, Peighambardoust SH, Golshan Tafti A, Hejazi MA, Rafat SA. Evaluating the ability of liquid sourdough containing L. plantarum and L. reuteri starters in inhibition of bread mold spoilage. J Food Res. 2011; 21(3): 391-399. [In Persian]
23. Vinderola CG, Reinheimer JA. Lactic acid starter and probiotic bacteria: a comparative in vitro study of probiotic characteristics and biological barrier resistance. Food Res Int. 2003; 36(9): 895-904.
24. Schillinger U, Guigas C, Holzapfel WH. In vitro adherence and other properties of lactobacilli used in probiotic yoghurt-like products. Int Dairy J. 2005; 15(2): 1289-1297.
25. Collado MC, Surono I, Meriluoto J, Salminen S. Indigenous dadih lactic acid bacteria: cell-surface properties and interactions with pathogens. J Food Sci. 2007; 72(3): 89-93.
26. Dzidic S, Suskovic J, Kos B. Antibiotic resistance mechanisms in bacteria: biochemical and genetic aspects. Food Technol Biotech. 2008; 46(1): 11-21.