مطالعه تعیین ویژگی‌های آنزیمی باکتری‌های شدیدا نمک دوست جداسازی شده از دریاچه حوض سلطان
محورهای موضوعی : میکروب شناسی
فهیمه محمودنیا
1
(استادیار، گروه زیست‌شناسی، دانشکده علوم، دانشگاه فرهنگیان، تهران، ایران)
کلید واژه: آنزیم‌های میکروارگانیسم‌ها, جداسازی باکتری‌ها, باکتری‌های نمک دوست, دریاچه نمک,
چکیده مقاله :
هدف:از دیرباز مطالعه بر روی میکروارگانیسم‌های مناطق ویژه با مشخصات خاص از اهمیت فراوانی برخوردار بوده است. دریاچه نمک حوض سلطان یکی از دریاچه‌های شور منطقه کویر مرکزی ایران می‌باشد که از نظر وسعت و میانگین شوری مورد توجه قرار گرفته است. تحقیق حاضر در جهت جداسازی باکتری‌های نمک دوست به منظور کسب اطلاعات بیشتر درباره تنوع میکروبی این دریاچه انجام شد. مواد و روش‌‌ها: از پنج منطقه دریاچه حوض سلطان نمونه‌گیری شد. نمونه‌ها رقیق‌سازی گردید و بر روی محیط مولتن هالوئیدآگار با غلظت‌های مختلفی از نمک (%35-5) کشت داده شدند. پلیت‌ها در دمای °C37 تحت شرایط هوازی انکوبه گردیدند. خصوصیات بیوشیمیایی، استفاده از منابع کربن و تولید اگزوآنزیم‌ها مورد بررسی قرار گرفت. یافته‌‌ها: 205 سویه در محیط‌های حاوی (%15- 5) نمک، رشد کردند. از بین سویه‌های جداسازی شده، 18 سویه در محیط حاوی %35- 15 غلظت نمک رشد داشتند که می‌توان آنها را به عنوان باکتری‌های شدیدا نمک دوست مورد توجه قرار داد. نتایج بدست آمده از بررسی‌های میکروسکوپی بیانگر آن است که 178سویه باسیل، کوکسی و اشکال رشته‌ای گرم مثبت بوده و 27 سویه باسیل گرم منفی می‌باشند. در شناسایی فنوتیپی سویه‌های ایزوله شده از باکتری‌های نمک دوست افراطی، جنس‌های هالوباکتریوم (Halobacterium)، هالوآرکئولا (Haloarcula)، هالوروبروم (Halorubrum) و هالوکوکوس (Halococcus) تشخیص داده شدند. علاوه براین، با بررسی تولید آنزیم توسط این سویه‌ها مشخص شد که برخی از آنها توانایی تولید آنزیم‌های متفاوتی مانند: آمیلاز، لیپاز، دی ان آز، اوره آز، گزیلاناز و ژلاتیناز را دارند. نتیجه‌‌گیری: دریاچه حوض سلطان از تنوع میکروب‌های نمک دوست وسیعی برخوردار است. علاوه براین، این باکتری‌ها می‌توانند به عنوان منبعی جهت تولید آنزیم‌های تحمل‌کننده نمک در صنایع گوناگون کاربردهای فراوانی داشته باشند.
Introduction: The study of microorganisms in specific regions with specific characteristics has long been important. Howz Soltan is a salt lake in the central desert zone of Iran, which is considered as an area of great salinity. The present study was conducted to isolate halophilic bacteria from Howz Soltan lake in order to achieve maximum information concerning to microbial diversity of the lake.Material and methods: For this purpose, samples were collected from five regions. Then the samples were diluted and cultivated on Molten haloid agar with different salt concentrations (5-35%). The plates were incubated at 37ºC in aerobic conditions. Biochemical characterizations, utilization of carbon sources and production of exoenzymes were investigated.Results: In total 205 different colonies were grew on medium with 5-35% salt concentrations. Of all isolates 18 strains were grew on medium with 15-35% salt concentrations. These strains were considered extreme halophilic bacteria and the rest were halotolerant and moderate halophilic bacteria. The results obtained from microscopic analysis of the isolates indicated that 178 isolates were gram positive bacilli, cocci and filamentous and 27 isolates were gram negative with bacilli shape. Phenotypic identification recognized that the isolated strains of extreme halophilic bacteria were Halobacterium ,Haloarcula ,Halorubrum and Halococcus. In addition, enzyme production assay of these strains showed some of them have capability to produce different enzymes viz., amylase, lipase, protease, DNase, urease, xylanase and gelatinase. Conclusion: In general, our finding showed the huge diversity of halophilic bacteria in Howz Soltan lake. Furthermore, these bacteria could be considered as sources of halotolerant enzymes in different industries.
Fendrihan S, Legat A, Pfaffenhuemer M, Gruber C, Weidler G, Gerbl F, Stan-Lotter H. Extremely halophilic archaea and the issue of long-term microbial survival. Rev. Environ. Sci. Biotechnol. 2006; 5: 203–218.
Dave SR, Desai HB. Microbial diversity at marine salterns near Bhavnagar, Gujarat, India. Current Science. 2016; 60(4): 497-500.
DasSharma S, Arora P. Halophiles, Encyclopedia of Life Sciences. Nature Publishing Group, 2011: 31-89.
Oren A. Halophilic microorganisms and their environments. Kluwer Academic Publisher, Dordrecht/Boston/London, The Netherlands, 2003: 30–74.
Van den Burg B. Extremophiles as a source for novel enzymes. Ecol.ind. microbial. 2013; 6: 213–218.
Wikipedia, the free encyclopedia, Namak Lake. 2017.
http://en.wikipedia.org/wiki/Namak-Lake.
Ventosa A, Nieto JJ, Oren A. Biology of moderately halophilic aerobic bacteria, Microbiol. Mol. Biol. Rev. 2012; 62: 504–544.
Enache M, Itoh T, Kamekura M, Teodosiu G, Dumitru L. Haloferax prahovense sp. nov.an extremely halophilic archaeon isolated from a Romanian salt lake. Int. J. Syst. Evol. Microbiol. 2007; 57: 393–397.
Gutiérrez MC, Castillo AM, Kamekura M, Xue Y, Ma Y, Cowan DA, Jones BE, Grant WD, Ventosa A. Halopiger xanaduensisgen. nov., sp. nov., an extremely halophilic archaeon isolated from saline Lake Shangmatala in Inner Mongolia, China. Int. J. Syst. Evol. Microbiol. 2007; 57: 1402–1407.
Jiang H, Dong H, Zhang G, Yu B, Chapman LR. Microbial diversity in water and sediment of Lake Chaka an athalassohaline lake in northwestern China. Appl. Environ. Microbiol. 2016; 72: 3832–3845.
Stan-Lotter H, Radax C, McGenity TJ, Legat A, Pfaffenhuemer M, Wieland H, Gruber C, Denner EBM. From intraterrestrials to extraterrestrials –viable haloarchaea in ancient salt deposits. In: text book of Halophilic microorganisms. edited by: Ventosa A, Springer Verlag, Berlin, Heidelberg, New York, 2004: 89–102.
Grant WD, Kamekura M, McGenity TJ, Ventosa A. The order Halobacteriales. In: text book of Bergey’s Manual of Systematic Bacteriology. edited by: DR Boone and RW Castenholz, 2thedition, 2011, 1: 294–334.
Murray RGE, Doetsch RN, Robinow CF. Determination and cytological light microscopy. In: text book of Method for General and molecular bacteriology. edited by: Gerhardt P, Murray RGE, Wood WA, Krieg NR. washington. DC: America society for microbiology, 2014: 121-141.
Cowan DA. Industrial enzymes in biotechnology. edited by: The science and the Business. eds Moses V, Cape RE. Reading; Harwood Academic Publishers, 2008: 311-340.
Onishi H, Mori T, Takechi S, Tani K, Kobayashi T, Kamekura M. Halophilic nuclease of a moderately halophilic bacillus sp: production, purification and characteristics. Appl. Environ. Microbiol. 2008; (45): 24-30.
Kamekura M. Extreme halophiles, In text book of The Biology of Halophilic Bacteria. edited by: Vreeland RH, Hochstein LI, Boca Raton: CRC Press, 2003: 135–161.
Sanchez-Porro C, Martin S, Mellado E, Ventosa A. Diversity of Moderately Halophilic Bacteria Producing Extracellular Hydrolytic Enzymes. Appl.Microbiol. 2003; 94: 295-300.
Kushner DJ, Kamekura M. Physiology of halophilic Eubacteria. In: text book of Halophilic Bacteria. edited by: Rodriguez-Valera F, CRC Press, Inc. Boca Raton, Florida, 2008, 1: 109–138.
Norton CF, McGenity TJ, Grant WD. Archaeal halophiles (halobacteria) from two British salt mines. J.Gen.Microbiol. 2013; 139: 1077–1081.
Post FJ. The microbial ecology of the Great Salt Lake. Microb.Ecol. 2004; 3:143–165.
Yang Y, Cui HL, Zhou PJ, Liu SJ. Haloarcula amylolytica sp.nov., an extremely halophilic archaeon isolated from Aibi salt lake in Xin-Jiang, China. Int. J.Syst. Evol. Microbiol. 2007; 57: 103–106.
Hughes JB, Hellmann JJ, Ricketts TH, Bohannan BJM. Counting the uncountable: statistical approaches to estimating microbial diversity. Appl Environ Microbiol 2001; 67(10): 4399-4406.
Babavalian H, Amoozegar M, Pourbabaei AA. Isolation, Identification and Characterization of moderately halophilic bacteria producing Hydrolytic enzymes from Aran-Bidgol salt Lake. Iranian Journ of Biology. 2009; 22(1): 24-45.
Mehrshad M, Amoozegar M, Yakhchali B, Shahzedeh Fazeli A. Biodiversity of moderately halophilic and halotolerant bacteria in the western coastal line of Urmia lake. Biological Journal of Microorganisms. 2012; 1(2): 49-70.
Zununi Vahed S, Forouhandeh H, Hassanzadeh S, Klenk H, Hejazi MA, Hejazi MS. Isolation and characterization of halophilic bacteria from Urmia Lake in Iran. Microbiology.2011; 80(6): 834–841.
Kafilzadeh F, Javid H, Kargar M. Isolation of Halophilic and Halotolerant Microorganisms from the Bakhtegan Lake and the Effect of Physicochemical Factors on Their Frequency. Journal of water and wastewater. 2007; 18(3): 81-87.
Zarparvar P, Amoozegar M, Fallahian M. Diversity of culturable Moderate halophilic and Halotolerant Bacteria in Incheh Boroun hyper saline wetland in Iran. Iranian Journal of Cellular and Molecular Researches. 2014; 1: 44-56.
Makhdoumi-Kakhki A, Amoozegar MA, Kazemi B, Pašić L, Ventosa A. Prokaryotic diversity in Aran-Bidgol salt lake, the largest hypersaline playa in Iran. Microbes Environ. 2011; 27(1): 87-93.
Alain K, Querellou J. Cultivating the uncultured: limits, advances and future challenges. Extremophiles. 2009; 13(4): 583-594.
_||_Fendrihan S, Legat A, Pfaffenhuemer M, Gruber C, Weidler G, Gerbl F, Stan-Lotter H. Extremely halophilic archaea and the issue of long-term microbial survival. Rev. Environ. Sci. Biotechnol. 2006; 5: 203–218.
Dave SR, Desai HB. Microbial diversity at marine salterns near Bhavnagar, Gujarat, India. Current Science. 2016; 60(4): 497-500.
DasSharma S, Arora P. Halophiles, Encyclopedia of Life Sciences. Nature Publishing Group, 2011: 31-89.
Oren A. Halophilic microorganisms and their environments. Kluwer Academic Publisher, Dordrecht/Boston/London, The Netherlands, 2003: 30–74.
Van den Burg B. Extremophiles as a source for novel enzymes. Ecol.ind. microbial. 2013; 6: 213–218.
Wikipedia, the free encyclopedia, Namak Lake. 2017.
http://en.wikipedia.org/wiki/Namak-Lake.
Ventosa A, Nieto JJ, Oren A. Biology of moderately halophilic aerobic bacteria, Microbiol. Mol. Biol. Rev. 2012; 62: 504–544.
Enache M, Itoh T, Kamekura M, Teodosiu G, Dumitru L. Haloferax prahovense sp. nov.an extremely halophilic archaeon isolated from a Romanian salt lake. Int. J. Syst. Evol. Microbiol. 2007; 57: 393–397.
Gutiérrez MC, Castillo AM, Kamekura M, Xue Y, Ma Y, Cowan DA, Jones BE, Grant WD, Ventosa A. Halopiger xanaduensisgen. nov., sp. nov., an extremely halophilic archaeon isolated from saline Lake Shangmatala in Inner Mongolia, China. Int. J. Syst. Evol. Microbiol. 2007; 57: 1402–1407.
Jiang H, Dong H, Zhang G, Yu B, Chapman LR. Microbial diversity in water and sediment of Lake Chaka an athalassohaline lake in northwestern China. Appl. Environ. Microbiol. 2016; 72: 3832–3845.
Stan-Lotter H, Radax C, McGenity TJ, Legat A, Pfaffenhuemer M, Wieland H, Gruber C, Denner EBM. From intraterrestrials to extraterrestrials –viable haloarchaea in ancient salt deposits. In: text book of Halophilic microorganisms. edited by: Ventosa A, Springer Verlag, Berlin, Heidelberg, New York, 2004: 89–102.
Grant WD, Kamekura M, McGenity TJ, Ventosa A. The order Halobacteriales. In: text book of Bergey’s Manual of Systematic Bacteriology. edited by: DR Boone and RW Castenholz, 2thedition, 2011, 1: 294–334.
Murray RGE, Doetsch RN, Robinow CF. Determination and cytological light microscopy. In: text book of Method for General and molecular bacteriology. edited by: Gerhardt P, Murray RGE, Wood WA, Krieg NR. washington. DC: America society for microbiology, 2014: 121-141.
Cowan DA. Industrial enzymes in biotechnology. edited by: The science and the Business. eds Moses V, Cape RE. Reading; Harwood Academic Publishers, 2008: 311-340.
Onishi H, Mori T, Takechi S, Tani K, Kobayashi T, Kamekura M. Halophilic nuclease of a moderately halophilic bacillus sp: production, purification and characteristics. Appl. Environ. Microbiol. 2008; (45): 24-30.
Kamekura M. Extreme halophiles, In text book of The Biology of Halophilic Bacteria. edited by: Vreeland RH, Hochstein LI, Boca Raton: CRC Press, 2003: 135–161.
Sanchez-Porro C, Martin S, Mellado E, Ventosa A. Diversity of Moderately Halophilic Bacteria Producing Extracellular Hydrolytic Enzymes. Appl.Microbiol. 2003; 94: 295-300.
Kushner DJ, Kamekura M. Physiology of halophilic Eubacteria. In: text book of Halophilic Bacteria. edited by: Rodriguez-Valera F, CRC Press, Inc. Boca Raton, Florida, 2008, 1: 109–138.
Norton CF, McGenity TJ, Grant WD. Archaeal halophiles (halobacteria) from two British salt mines. J.Gen.Microbiol. 2013; 139: 1077–1081.
Post FJ. The microbial ecology of the Great Salt Lake. Microb.Ecol. 2004; 3:143–165.
Yang Y, Cui HL, Zhou PJ, Liu SJ. Haloarcula amylolytica sp.nov., an extremely halophilic archaeon isolated from Aibi salt lake in Xin-Jiang, China. Int. J.Syst. Evol. Microbiol. 2007; 57: 103–106.
Hughes JB, Hellmann JJ, Ricketts TH, Bohannan BJM. Counting the uncountable: statistical approaches to estimating microbial diversity. Appl Environ Microbiol 2001; 67(10): 4399-4406.
Babavalian H, Amoozegar M, Pourbabaei AA. Isolation, Identification and Characterization of moderately halophilic bacteria producing Hydrolytic enzymes from Aran-Bidgol salt Lake. Iranian Journ of Biology. 2009; 22(1): 24-45.
Mehrshad M, Amoozegar M, Yakhchali B, Shahzedeh Fazeli A. Biodiversity of moderately halophilic and halotolerant bacteria in the western coastal line of Urmia lake. Biological Journal of Microorganisms. 2012; 1(2): 49-70.
Zununi Vahed S, Forouhandeh H, Hassanzadeh S, Klenk H, Hejazi MA, Hejazi MS. Isolation and characterization of halophilic bacteria from Urmia Lake in Iran. Microbiology.2011; 80(6): 834–841.
Kafilzadeh F, Javid H, Kargar M. Isolation of Halophilic and Halotolerant Microorganisms from the Bakhtegan Lake and the Effect of Physicochemical Factors on Their Frequency. Journal of water and wastewater. 2007; 18(3): 81-87.
Zarparvar P, Amoozegar M, Fallahian M. Diversity of culturable Moderate halophilic and Halotolerant Bacteria in Incheh Boroun hyper saline wetland in Iran. Iranian Journal of Cellular and Molecular Researches. 2014; 1: 44-56.
Makhdoumi-Kakhki A, Amoozegar MA, Kazemi B, Pašić L, Ventosa A. Prokaryotic diversity in Aran-Bidgol salt lake, the largest hypersaline playa in Iran. Microbes Environ. 2011; 27(1): 87-93.
Alain K, Querellou J. Cultivating the uncultured: limits, advances and future challenges. Extremophiles. 2009; 13(4): 583-594.
