| تعداد نشریات | 282 |
| تعداد نشریات سازمان | 80 |
| تعداد شمارهها | 3,208 |
| تعداد مقالات | 35,560 |
| تعداد مشاهده مقاله | 53,002,435 |
| تعداد دریافت فایل اصل مقاله | 92,287,874 |
افزایش تولید سورفکتین در باکتری Bacillus subtilis UTB96 با استفاده از بهینهسازی فرایند فرمانتاسیون | ||
| مهار زیستی در گیاهپزشکی | ||
| مقاله 2، دوره 5، شماره 2 - شماره پیاپی 10، اسفند 1396، صفحه 13-26 اصل مقاله (341.18 K) | ||
| نوع مقاله: مقاله پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/bcpp.2018.117885 | ||
| نویسندگان | ||
| سحر کشاورزی1؛ مسعود احمدزاده1؛ سعید میرزایی2؛ کیوان بهبودی1؛ مژگان بندهپور بندهپور3 | ||
| 1گروه گیاهپزشکی، پردیس کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران | ||
| 2گروه بیوتکنولوژی کشاورزی، دانشگاه تحصیلات تکمیلی صنعتی و فناوری پیشرفته کرمان، کرمان، ایران | ||
| 3دانشکده فناوریهای نوین پزشکی، دانشگاه علوم پزشکی شهید بهشتی، تهران، ایران | ||
| چکیده | ||
| در میان باکتریهای زیست مهارگر، باکتری Bacillus subtilis در تولید طیف وسیعی از ترکیبات ضدمیکروبی مطرح میباشد که از این میان، بیوسورفکتانتها از جمله سورفکتین از اهمیت ویژهای برخورد است. سورفکتانتهایی که بهوسیلۀ میکروارگانیسمها تولید میشوند بهدلیل سمیّت پایین و خواص منحصر به فرد، کاربردهای متعددی دارند اما بهدلیل هزینه بالای تولید و خالصسازی، تولید در بازده بالا هنوز میسر نشده است. در این تحقیق بهمنظور ارزیابی تولید سورفکتین در جدایه مذکور، شناسایی ژن تولید کننده آن (srf) و تعیین مقدار سورفکتین تولید شده بهترتیب از روشهای متلاشی شدن قطره (Drop collapse)، روش مولکولی واکنش زنجیرهای پلیمراز (PCR) و آنالیز کروماتوگرافی مایع با کارایی بالا (HPLC) استفاده شد. بررسی مولکولی با آغازگر اختصاصی نشان داد که این جدایه دارای ژنsrf میباشد. برای بهینهسازی شرایط عملیاتی، اثر شرایط فیزیک وشیمیایی و القا کنندههای مختلف بر میزان زیست توده و سورفکتین خام تولیدی بررسی شد. شرایط بهینه فیزیکی و شیمیایی بهدست آمده شامل: مدّت زمان نگهداری کشت 96 ساعت، دمای 30 درجۀ سلسیوس، اسیدیته 7 و دور شیکر rpm 250 میباشد. از میان القاکنندهها که شامل منابع کربن، منابع نیتروژن و اسیدهای آمینه، بیشترین مقدار تولید سورفکتین پس از تیمار محیط کشت با گلوکز بهمقدار 243/1 گرم بر لیتر محیط کشت میباشد. پس از آن تأثیر عناصر القاکنندهای که بیشترین تأثیر را در افزایش تولید سورفکتین داشتهاند، بهصورت توأم بررسی شد. نتایج نشان داد بیشترین مقدار تولید سورفکتین mg/l 055/5 در حضور یونهای آهن، منیزیم و منگنز بهصورت توأم میباشد. همچنین مقدار و نوع منابع غذایی تأثیر بهسزایی در بازده تولید سورفکتین دارد. بنابر این تغییر در مقدار و ترکیب القا کنندهها روشی اقتصادی برای تولید تجاری این ماده محسوب میشود. | ||
| کلیدواژهها | ||
| سورفکتانت؛ فرمانتاسیون؛ ژن srf؛ کروماتوگرافی مایع با کارایی بالا | ||
| عنوان مقاله [English] | ||
| Enhancing surfactant production in Bacillus subtilis UTB96 by fermentation optimization | ||
| نویسندگان [English] | ||
| Sahar Keshavarzi1؛ Masoud Ahmad zadeh1؛ Saeid Mirzaei2؛ Keivan Behboudi1؛ Mojgan bandeh pour3 | ||
| 2Department of Biotechnology, Graduate University of Advanced Technology, Kerman, Iran | ||
| 3Biotechnology Department, School of Advacned Technologies in Medicine, Shahid Beheshti,University of Medical Sciences, Tehran, Iran | ||
| چکیده [English] | ||
| A biocontrol bacterium, Bacillus subtilis is well known for producing wide spectrum of antimicrobial compounds. Among these products, biosurfactant compounds such as surfactin have high importance. Biosurfactants that are produced by the microorganisms are applied in the various aspects of industry, medicine and agriculture because of their low toxicity and individual properties. However, these compounds are not produced in high quantity due to their low production yield and high purification cost. In the current study for investigating the production of surfactin in the isolate (UTB96), recognition of srf gene and determination of the amount of surfactin production, we used Drop collapse, PCR and HPLC methods. Molecular analysis with specific primer indicated that the isolate had srf gene. To optimize the fermentation process, influence of various inducers and physiochemical factors were studied on the biomass and surfactin production. The results indicated that maximum surfactin production occurred after amending the culture medium with glucose (1.243 g/l) solution. After that influence of the combined inducer elements were studied in increasing the amount of surfactin production. Results revealed that maximum production (5.055 g/l) was done in the presence of Fe2+, Mn2+ and Mg2+ mixture in the culture medium. The results also showed that the amount and the type of the food sources had remarkable impact on the surfactin production efficiency. The changes in the amount and composition of the food sources therefore, could be economically considered for the commercial production of this compound. | ||
| کلیدواژهها [English] | ||
| biosurfactant, fermentation, srf gene, HPLC | ||
| مراجع | ||
|
Abdel-Mawgod, A., Aboulwafa, M. & Hassouna, N. 2008. Characterizations of surfactin produced by Bacillus subtilis isolate BS5. BiotechnologyandApplied Biochemistry, 150: 289–303. Akpa, E., Jacques P., Wathelet, B., Paquot, M., Fuchs, R., Budzikiewicz, H. & Thonart, P. 2001. Influence of culture conditions on lipopeptide production by Bacillus subtilis. Applied Biochemistryand Biotechnology, 93: 551–561. Al-Ajlani, M.M., Sheikh, M.A., Ahmad, Z. & Hasnain, S. 2007. Production of surfactin from Bacillus subtilis MZ-7 grown on pharmamedia commercial medium. Microbial Cell Factories, 6: 17. Arima, K., Kakinuma, A. & Tamura, G. 1968. Surfactin, a crystalline peptidelipid surfactant produced by Bacillus subtilis: isolation, characterization and its inhibition of fibrin clot formation. Biochemicaland Biophysical Research Communications, 31: 488–494. Askari, N., Salehi Jouzani, Gh., Mousivand, M., Foroutan, A., Hagh Nazari, A., Abbasalizadeh, S., Soheilivand, S. & Mardi, M. 2011. Evaluation of Anti-Phytoplasma Properties of Surfactin and Tetracycline Towards Lime Witches’ Broom Disease Using Real-Time PCR. Journal of MicrobiologyandBiotechnology, 21: 81–88. Assie, L.K., Deleu, M., Arnaud, L., Paquot, M., Thonart, P., Gaspar, Ch. & Haubruge, E. 2002. Insecticide activity of surfactins and iturins from a biopesticide Bacillus subtilisCohn (S499 strain). Meded. Rijksuniv. Gent. Fak. Landbouwkd. Toegep. Biol. Wet., 67: 647–655. Bais, H.P., Fall, R. & Vivanco, J.M. 2004. Biocontrol of Bacillus subtilis against infection of Arabidopsis roots by Pseudomonas syringae is facilitated by biofilm formation and surfactin. Plant Physiology, 134: 307–319. Cooper, D.G., Macdonald, C.R., Duff, S.J. & Kosaric, N. 1981. Enhanced production of surfactin from Bacillus subtilis by continuous product removal and metal cation additions. AppliedandEnvironmental Microbiology, 42: 408–412. Cosby, W.M., Vollenbroich, D., Lee, O.H. & Zuber, P. 1998. Altered srf expression in Bacillus subtilis resulting from changes in culture pH is dependent on the Spo0K oligopeptide permease and theComQX system of extracellular control. Journal of Bacteriology, 180: 1438–1445. Coutte, F., Lecle`re, V., Be´chet, M., Guez, J.S., Lecouturier, D., Chollet Imbert, M., Dhulster, P. & Jacques, P. 2010. Effect of pps disruption and constitutive expression of srfA on surfactin productivity, spreading and antagonistic properties of Bacillus subtilis 168 derivatives. Journal of Applied Microbiology, 109: 480–491. Debois, D., Hamze, K., Gue´rineau, V., Le Cae¨r, J.P., Holland, I.B., Lopes, P., Ouazzani, J., Se´ror, S.J., Brunelle, A. & Lapre´vote, O. 2008. In situ localisation and quantification of surfactins in a Bacillus subtilis swarming community by imaging mass spectrometry. Proteomics, 8: 3682–3691. Gancel, F., Montastruc, L., Liu, T., Zhao, L. & Nikov, I. 2009. Lipopeptide overproduction by cell immobilization on iron enriched light polymer particles. Process Biochemistry, 44: 975–978. Gevers, W., Kleinkauf, H. & Lipmann, F. 1968. The activation of amino acids for biosynthesis of gramicidin S. National Academy Sciences, 60: 269–276. Hsieh, F.C., Li, M.C., Lin, T.C. & Kao, S.S. 2004. Rapid detection and characterization of surfactinproducing Bacillus subtilis and closely related species based on PCR. Current Microbiology, 49: 186–191. Jacques, P., Hbid, C., Destain, J., Razafindralambo, H., Paquot, M., De Pauw, E. & Thonart, P. 1999. Optimization of biosurfactant lipopeptide production from Bacillus subtilis S499 by Plackett Burman design. Appleid Biochemistryand Biotechnology, 77: 223–233. Kim, H.S., Yoon, B.D., Lee, C.H., Suh, H.H., Oh, H.M., Katsuragi T. & Tani Y. 1997. Production and properties of a lipopeptide biosurfactant from Bacillus subtilis C9. Journal ofFermentationTechnology, 84: 41–46. Kinsinger, R.F., Shirk, M.C. & Fall, R. 2003. Rapid surface motility in Bacillus subtilis is dependent on extracellular surfactin and potassium ion. Journal of Bacteriology, 185: 5627–5631. Kinsinger, R.F., Kearns, DB., Hale, M. & Fall, R. 2005. Genetic requirements for potassium ion-dependent colony spreading in Bacillus subtilis. Journal of Bacteriology, 187: 8462–8469. Konz, D., Klens, A., Schorgendorfer, K. & Marahiel, M.A. 1997. The bacitracin biosynthesis operon of Bacillus licheniformis ATCC 10716: molecular characterization of three multi-modular peptide synthetases. Chemical Biology, 4: 927–937. Kracht, M., Rokos, H., Ozel, M., Kowall, M., Pauli, G. & Vater, J. 1999. Antiviral and hemolytic activities of surfactin isoforms and their methyl ester derivatives. Journal of Antibiotics, 52: 613–619. Landy, M., Warren, G.H., Rosenman, S.B. & Colio, L.G. 1948. Bacillomycin an antibiotic from Bacillus subtilis active against pathogenic fungi. Proceedings of the Societyfor Experimental Biologyand Medicine, 67: 530–541. Mohammadipour, M., Mousivand, M., Salehi Jouzani, Gh. & Abbasalizadeh, S. 2009. Molecular and biochemical characterization of Iranian surfactin-producing Bacillus subtilis isolates and evaluation of their biocontrol potential against Aspergillus flavus and Colletotrichum gloeosporioides.Canadian Journal of Microbiology, 55: 395–404. Mootz, H.D., Kessler, N., Linne, U., Eppelmann, K., Schwarzer, D. & Marahiel, M.A. 2002. Decreasing the ring size of a cyclic no ribosomal peptide antibiotic by in frame module deletion in the biosynthetic genes. Journal of the American Chemical Society, 124: 10980–10981. Mukherjee, S., Das, P. & Sen, R. 2006. Towards commercial production of microbial surfactants.Trends in Biotechnology, 24: 509–515. Muthusamy, K., Gopalakrishnan, S., Ravi, T.K. & Sivachidambaram, P. 2008. Biosurfactants: properties, commercial production and application. Current Science, 94: 736–747. Nagai, S., Okimura, K., Kaizawa, N., Ohki, K. & Kanatomo, S. 1996. Study on surfactin, a cyclic depsipeptide. II. Synthesis of surfactin B2 produced by Bacillus natto KMD 2311. Chemical andPharmaceuticalBulletin, 44: 5–10. Ohno, A., Ano, T. & Shoda, M. 1992. Production of a lipopeptide antibiotic surfactin with recombinant Bacillus subtilis. Biotechnology Letters, 14: 1165–1168. Ohno, A., Ano, T. & Shoda, M. 1995. Production of a lipopeptide antibiotic, surfactin, by recombinant Bacillus subtilis in solid state fermentation. Biotechnology and Bioengineering, 47: 209–214. Ongena, M., Jacques, P., Toure´, Y., Destain, J., Jabrane, A. & Thonart, P. 2005. Involvement of fengycin type lipopeptides in the multifaceted biocontrol potential of Bacillus subtilis. Applied Microbiology andBiotechnology, 69: 29–38. Ongena, M. & Jacques, P. 2008. Bacillus lipopeptides: versatile weapons for plant disease biocontrol. Trends inMicrobiology, 16: 115–125. Ongena, M., Jourdan, E., Adam, A., Paquot, M., Brans, A., Joris, B., Arpigny, J.L. & Thonart, P. 2007. Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants. Environmental Microbiology, 9: 1084–1090. Peypoux, F., Bonmatin, J.M. & Wallach, J. 1999. Recent trends in the biochemistry of surfactin. AppliedMicrobiology andBiotechnology, 51: 553–563. Rashedi, ,H.R., Mazaheri Assadi, M., Jamshidi, E. & Bonakdarpour, B. 2006. Optimization of the production of biosurfactant by Pseudomonas aeruginosa HR isolated from an Iranian Southern Oil well. Iranian Journal of Chemistry and Chemical Engineering, 25: 25–30. Ritter, S.K. 2004. Green innovations. Chemical & Engineering News, 82: 25–30. Sandrin, C., Peypoux, F. & Michel, G. 1990. Coproduction of surfactin and iturin A, lipopeptides with surfactant and antifungal properties, by Bacillus subtilis. Biotechnology and Applied Biochemistry, 12: 370–375. Sen, R. & Swaminathan, T. 1997. Application of response-surface methodology to evaluate the optimum environmental conditions for the enhanced production of surfactin. Applied Microbiology andBiotechnology,47:358–363. Shaligram, N.S. & Singhal, R.S. 2010. Surfactin: A review on biosynthesis, fermentation, purification and applications. Food Technology and Biotechnology, 48: 119–134. Sheppard, J.D. & Cooper, D.G. 1991. The response of Bacillus subtilis ATCC 21332 to manganese during continuous phased growth. Applied Microbiology and Biotechnology, 35: 72–76. Sun, H., Bie, X., Lu, F., Lu, Y., Wu, Y. & Lu, Z. 2009. Enhancement of surfactin production of Bacillus subtilis fmbR by replacement of the native promoter with the Pspac promoter. Canadian Journal of Microbiology,5: 1003–1006. Thomas, D.W. & Ito. T. 1969. The revised structure of the peptide antibiotic esperin, established by mass spectrometry. Tetrahedron, 25: 1985–1990. Tugrul, T. & Cansunar, E. 2005. Detecting surfactant-producing microorganisms by the drop-collapse test. Journal of Microbiology and Biotechnology, 21: 851–853. Vollenbroich, D., Pauli, G., Ozel, M. & Vater, J. 1997. Antimycoplasma properties and application in cell culture of surfactin, a lipopeptide antibiotic from Bacillus subtilis. Applied and Environmental Microbiology, 63: 44–49. Wei, Y.H. & Chu, I.M. 2002. Mn2+ improves surfactin production by Bacillus subtilis. Biotechnology Letters, 24: 479–482. Wei, Y.H., Lai, C.C. & Chang, J.S. 2007. Using Taguchi experimental design methods to optimize trace element composition for enhanced surfactin production by Bacillus subtilis ATCC 21332. Process Biochemistry, 42: 40–45. Wei, Y.H., Wang, L.F. & Chang, J.S. 2004. Optimizing iron supplement strategies for enhanced surfactin production with Bacillus subtilis. Biotechnology Progress, 20: 979–983. Yeh, M.S., Wei, Y.H. & Chang, J.S. 2005. Enhanced production of surfactin from Bacillus subtilis by addition of solid carriers. Biotechnology Progress, 21:1329–1334. | ||
|
آمار تعداد مشاهده مقاله: 1,151 تعداد دریافت فایل اصل مقاله: 667 |
||