بررسی تولید روغن تک یاخته در مراحل مختلف رشد در مخمر مولد چربی ردوترولا موسیلوژینوزا سویه UIMC35 با تکنیک فلوسایتومتری

نوع مقاله: مقاله پژوهشی

نویسندگان

1 استادیار ، بخش میکروبیولوژی، گروه زیست شناسی، دانشکده علوم، دانشگاه اصفهان

2 استاد، بخش میکروبیولوژی، گروه زیست شناسی، دانشکده علوم، دانشگاه اصفهان

چکیده

در تحقیق حاضر مخمر بومی مولد چربی به نام Rhodotorula mucilaginosa سویه UIMC35 جهت تولید روغن تک یاخته جداسازی و شناسایی گردید. تکنیک فلوسایتومتری جهت بررسی میزان شدت فلورسنت سلولی (در کانال FL2)، اندازه سلولی (با پارامتر FSC) و تراکم درون سلولی (با پارامتر SSC) به کار گرفته شد. رابطه بین شدت فلورسنت و محتوای لیپیدی، جهت محاسبه سریع و آسان میزان محتوای لیپیدی ارائه گردید. تغییرات فاکتورهای مختلف همچون میزان تولید لیپید و بیومس، محتوای لیپیدی، کارایی تولید لیپید و بیومس، میزان قند، میزان ازت، نسبت کربن به ازت و pH در مراحل مختلف رشد مخمر در کشت بسته مورد بررسی قرار گرفت. بیشترین میزان تولید لیپید، محتوای لیپیدی و کارایی تولید لیپید در مخمر مورد بررسی به ترتیب 57/6 گرم در لیتر، 04/67 درصد و 13/21 درصد بدست آمد. کاهش نسبت C/N در مراحل انتهایی کشت از عوامل تاثیر گذار در کاهش میزان تولید لیپید به شمار می رود.

کلیدواژه‌ها

عنوان مقاله [English]

Monitoring lipid production in growth phases of new isolated oleaginous yeast Rhodotorula mucilaginosa strain UIMC35 by flow cytometry

نویسندگان [English]

  • Hossein Ghanavati 1
  • Iraj Nahvi 2

چکیده [English]

In present study, oleaginous yeast Rhodotorula mucilaginosa strain UIMC35 was isolated and identified for SCO (single Cell Oil) production. Flow cytometric technique was employed for the monitoring of fluorescence intensity of the cells (channel FL2), cell size (by FSC parameter) and the cell granularity (by SSC parameter). The relationship between fluorescence intensity and lipid content in order to quick and easy calculation of the amount of lipid content was presented. Changes in various factors such as lipid and biomass production, lipid content, lipid and biomass production efficiency, the amount of sugar, the amount of nitrogen, C/N ratio and pH were investigated at different stages of yeast strain batch culture. The highest rates of lipid production, lipid content and lipid production efficiency by this yeast strain were achieved 6.57 g/l, 67.04 % and 21.13 %, respectively. Decreasing of C/N ratio in the later stages of yeast growth caused the reduction of the lipid production rate.

کلیدواژه‌ها [English]

  • Oleaginous yeast
  • Flow cytometry
  • Fluorescent intensity
  • Lipid content
  • Subpopulation

 

Andrade, R. Lea, R. Roseiro, J. Reis, A. Lopes da Silva, T. (2012)  Monitoring Rhodosporidium toruloides NCYC 921 batch fermentations growing under carbon and nitrogen limitation by flow cytometry. World Journal of Microbiology & Biotechnology  28:1175-1184.

Clesceri, L.S. Greenberg, A.E. Eaton, A.D. (1999) Standard methods for the examination of water and wastewater, 20th ed., American Public Health Association: Washington DC.

Dai, C. Tao, J. Xie, F. Dai, Y. Zhao, M. (2007) Biodiesel generation from oleaginous yeast Rhodotorula glutinis with xylose assimilating capacity. African Journal of Biotechnology 6: 2130-2134.

He, M. Hu, Q. Gou, X. Liu, X. Li, Q. Pan, K. Zhu, Q. Wu, J. (2010) Screening of oleaginous yeast with xylose assimilating capacity for lipid and bio-ethanol production. African Journal of Biotechnology 9(49): 8392-8397.

Kurtzman, C.P. Fell, J.W. (2000) The yeasts: a taxonomic study, 4th ed., Elsevier, Amsterdam.  

Li, Q. Du, W. Liu, D. (2008) Perspectives of microbial oils for biodiesel production. Applied Microbiology & Biotechnology 80:749-756.

Li, Y. Zhao, Z. Bai, F. (2007) High-density cultivation of oleaginous yeast Rhodosporidium toruloides Y4 in fed-batch culture. Enzyme Microbial Technology 41: 312-317.

Lopes da Silva, T. Carlos Roseiro, J. Reis, A. (2012) Applications and perspectives of multi-parameter flow cytometry to microbial biofuels production processes. Trends in Biotechnology 30(4): 225-232.

Lopes da Silva, T. Feijão, D. Carlos Roseiro, J. Reis, A. (2011) Monitoring Rhodotorula glutinis CCMI 145 physiological response and oil production growing on xylose and glucose using multi-parameter flow cytometry. Bioresource Technology 102: 2998-3006.

Lopes da Silva, T. Feijão, D. Reis, A. (2010) Using multi-parameter flow cytometry to monitor the yeast Rhodotorula glutinis CCMI 145 batch growth and oil production towards biodiesel. Applied Biochemistry & Biotechnology 162: 2166-2176.

Miller, G.L. (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugars. Annals of Chemistry 31: 426-428.

Pan, L. Yang, D. Shao, L. Li, W. Chan, G. Liang, Z. (2009) Isolation of oleaginous yeasts from the soil as studies of their lipid-producing capacities. Food Technology & Biotechnology 47: 215-220.

Papanikolaou, S. Aggelis, G. (2011) Lipids of oleaginous yeasts. Part I: Biochemistry of single cell oil production. European Journal of Lipid Science & Technology 113, 1031–1051.

Raschke, D. Knorr, D. (2009) Rapid monitoring of cell size, vitality and lipid droplet development in the oleaginous yeast Waltomyces lipofer. Journal of Microbiological Methods 79: 178-183.

Shakeri, S. Roghanian, R. Emtiazi, G. (2011) Surveillance of single-cell behavior in different subpopulations of Ralstonia pickettii AR1 during growth and polyhydroxybutyrate production phases by flow cytometry. Journal of Basic Microbiology 51: 1–10.

Shakeri, S. Roghanian, R. Emtiazi, G. (2011) Comparison of intracellular polyhydroxybutyrate granules formation between different bacterial cell subpopulations by flow cytometry. Jundishapur Journal of Microbiology 4(4): 229-238.

White, T.J. Bruns, T. Lee, S. (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. p 315-322. in: M.A. Innis, D.H. Gelfand, J.J. Sninsky, T.J. White, (eds). PCR protocols: a guide to methods and applications, New York: Academic Press Inc.

Yamada, Y. Makimura, K. Mirhendi, H. Ueda, K. Nishiyama, Y. Yamaguchi, H. Osumi, M. (2002) Comparison of different methods for extraction of mitochondrial DNA from human pathogenic yeasts. Japanese Journal of Infection Disease 55:122-125.

Zhao, X. Kong, X. Hua, Y. Feng, B. Zhao, Z. (2008) Medium optimization for lipid production through co-fermentation of glucose and xylose by the oleaginous yeast Lipomyces starkeyi. European Journal of Lipid Science & Technology 110: 405-412.