Isolation and Screening of Oleaginous Yeasts Capable of Using Glycerol as a Carbon Source

Main Article Content

Patcharanan Amornrattanapa
Panisara Thongthep


Oleaginous yeasts are potential lipid producers that could be used for biodiesel production. In this present study, leaf and soil samples from a mangrove forest in Chonburi Province were collected and used for the isolation of yeasts in YEPG broth containing 2% glycerol as a carbon source in shaking flasks. The total of 26 isolates of yeast were obtained and they were further investigated for their ability to accumulate lipid by Sudan Black B staining. The result showed that 7 out of 26 yeast isolates; 1A 2C 3E 5B 6A 8A and 9A had intracellular lipid droplets. All of these isolates were estimated for their approximate amount of lipid accumulation by Nile Red fluorescence assay and only one isolate namely 1A that was isolated from leaves of Avicennia officinalis expressed the strongest potential for high level of lipid accumulation. After kinetic studies of biomass and lipid production of isolate 1A at 240 hours of growth in GMY broth, 24.24 ± 0.06 g/L of biomass yield, 3.63 ± 0.04 g/L of lipid yield, and lipid content of 14.98% of dry biomass were significantly achieved (p < 0.05). Strategies for improvement on lipid production from this isolate is needed to be further investigated prior to its utilization in the future.


Download data is not yet available.

Article Details

How to Cite
Amornrattanapa, P., & Thongthep, P. (2019). Isolation and Screening of Oleaginous Yeasts Capable of Using Glycerol as a Carbon Source. Ramkhamhaeng Research Journal of Sciences and Technology, 22(2), 61–70. Retrieved from
Author Biographies

Patcharanan Amornrattanapa, Faculty of Science, Burapha University

Lecturer at Department of Microbiology, Faculty of Science, Burapha University

Panisara Thongthep, Faculty of Science, Burapha University

Student at Department of Microbiology, Faculty of Science, Burapha University


Amaretti, A., Raimondi, S., Sala, M., Roncaglia, L., De Lucia, M., Leonardi, A. and Rossi, M. 2010. Single cell oils of the cold-adapted oleaginous yeast Rhodotorula glacialis DBVPG 4785. Microb. Cell Fact. 9: 73.
Burdon, K. 1946. Fatty material in bacteria and fungi revealed by staining dried, fixed slide preparations. J. Bacteriol. 52(6): 665-678.
Castanha, R. F., Morais, L. A. S. D., Mariano, A. P. and Monteiro, R. T. R. 2013. Comparison of two lipid extraction methods production methods produced by yeast in cheese whey. Braz. Arch. Biol. Technol. 56(4): 629-636.
Castrillón, M. R., Garcia, V. P. J., Rosa, P. D., Landell, M. F., Vu, D., Fabricio, M., Ayub, M. A. Z., Robert, V., Henriques, J. A. P. and Valente, P. 2017. The oleaginous yeast Meyerozyma guilliermondii BI281A as a new potential biodiesel feedstock: Selection and lipid production optimization. Front. Microbiol. 1776(8): 1-11.
Cuimin, H., Xin, Z., Jin, Z., Siguo, W. and Zongbao, K. Z. 2009. Effects of biomass hydrolysis by-products on oleaginous yeast Rhodosporidium toruloides. Bioresour. Technol. 100: 4843-4847.
Eamcharoen, A. and Rungrojchaipon, P. 2016. The conversion of glycerol to fuel additives. J. Sci. Ladkrabang. 25(2): 65-83.
Evan, C. T. and Ratledge, C. 1984. Influence of nitrogen metabolism on lipid accumulation in oleaginous yeasts. J. Gen. Microbiol. 130: 1693-1704.
Gientka, I., Gadaszewska, M., Blazejak, S., Kieliszek, M., Wrobel, B, A., Rozanska, S. L. and Kot, M. A. 2017. Evaluation of lipid biosynthesis ability by Rhodotorula and Sporobolomyces strains in medium with glycerol. Eur. Food Res. Technol. 243: 275-286.
Jape, A., Harsulkar, A. and Sapre, V. R. 2014. Modified Sudan Black B staining method for rapid screening of oleaginous marine yeasts. Int. J. Curr. Microbiol. Appl. Sci. 3(9): 41-46.
Jiru, T. M., Abate, D., Kiggundu, N., Pohl, C. and Groenewald, M. 2016. Oleaginous yeasts from Ethiopia. AMB Express. 6(1):78.
Kitcha, S. and Cheirsilp, B. 2011. Screening of oleaginous yeasts and optimization for lipid production using crude glycerol as a carbon source. Energy Procedia. 9: 274-282.
Kunthipun, S., Chokreansukchai, P., Hondee, P., Tanasupawat, S. and Savarajara, A. 2018. Diversity and characterization of cultivable oleaginous yeasts isolated from mangrove forests. World J. Microbiol. Biotechnol. 34: 125.
Li, Q., Du, W. and Liu, D. 2008. Perspectives of microbial oils for biodiesel production. Appl. Microbiol. Biotechnol. 80: 749-756.
Munch, G., Sestric, R., Sparling, R., Levin, B. D. and Cicek, N. 2015. Lipid production in the undercharacterized oleaginous yeasts, Rhodosporidium babjevae and Rhodosporidium diobovatum, from biodiesel-derived waste glycerol. Bioresour. Technol. 185: 49-55.
Papanikolaou, S. and Aggelis, G. 2011. Lipids of oleaginous yeasts. Part I: Biochemistry of single cell oil production. Eur. J. Lipid Sci. Tech. 113: 1031-1051.
Rakicka, M., Lazar, Z., Dulermo, T., Fickers, P. and Nicaud, J. M. 2015. Lipid production by the oleaginous yeast Yarrowia lipolytica using industrial by-products under different culture conditions. Biotechnol. Biofuels. 8: 104.
Santibañez, C., Varnero, M. T. and Bustamante, M. 2011. Residual glycerol from biodiesel manufacturing, waste or potential source of bioenergy: A review. Chil. J. Agr. Res. 71(3): 469-475.
Sitepu, I. R., Garay, L. A., Sestric, R., Levin, D., Block D. E., German, J. B. and Boundy-Mills, K. L. 2014. Oleaginous yeasts for biodiesel: Current and future trends in biology and production. Biotechnol. Adv. 32(7): 1336-1360.
Sitepu, I. R., lgnatia, L., Franz, A. K., Wong, D. M., Faulina, S. A., Tsui, M., Kanti, A. and Boundy-Mills, K. 2012. An improved high-throughput Nile red fluorescence assay for estimating intracellular lipids in a variety of yeast species. J. Microbiol. Meth. 91: 321-328.
Tapia, E. V., Anschau, A. A., Coradini, A. L., and Deckmann, A. C. 2012. Optimization of lipid production by the oleaginous yeast Lipomyces starkeyi by random mutagenesis coupled to cerulenin screening. AMB Express. 2: 64.