* Corresponding Author
UV Mutagenesis as a Strategy to Enhance Growth and Lipid Productivity of Chlorella sp. 042
https://doi.org/10.22146/jtbb.56862 Rike Rachmayati(1*), Eva Agustriana(2), Delicia Yunita Rahman(3)
(1) Research Center for Biotechnology, Indonesian Institute of Science
(2) Research Center for Biotechnology, Indonesian Institute of Science
(3) Research Center for Biotechnology, Indonesian Institute of Science
(*) Corresponding Author
Abstract
Microalgae appeared to be an alternative feedstock for renewable biodiesel production due to their capability to accumulate considerable amounts of lipids. In this study, mutagenesis using UVC light with different periods was applied to Chlorella sp. 042 to produce a microalgae strain with high lipid productivity of 45, 60, and 75 min. The Nile red fluorescence method was conducted to select a Chlorella sp. mutant with high neutral lipid and generated one mutant from every UV mutation period, M45-06, M60-02, and M75-21. All of the mutants have higher growth rates than the wild type. Chlorella sp. 042 M60-02 achieved the highest lipid productivity, with 34 mg L-1 day-1. Furthermore, as other major biochemical components, carbohydrate and protein contents were determined. Our results showed that all the mutants enhance their carbohydrate and protein contents compared to the wild type. However, mutations for more than 60 min do not intensely change the protein content of mutant microalgae. Gas chromatography-mass spectrophotometry analysis revealed that M60-02 mutant has similar FAME profiles with the wild type, which contain palmitic acid (C16:0), stearic acid (C 18:0), oleic acid (C18:1), and linoleic acid (C18:2). These results demonstrate that the UV mutation of Chlorella sp. 042 for 60 min is suitable as a source of biodiesel production.
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Abbaszaadeh, A., Ghobadian, B., Omidkhah, M. R. and Najafi, G., 2012, Current biodiesel production technologies: a comparative review, Energy Conversion and Management 63, 138–148.
Abu Sepian, N. R., Mat Yasin, N. H., Zainol, N., Rushan, N.H. and Ahmad, A.L., 2017, Fatty acid profile from immobilised Chlorella vulgaris cells in different matrices, Environmental Technology 40(9), 1110–1117.
Ahmad, A. L., Yasin, N. H. M., Derek, C. J. C. and Lim, J. K., 2011, Microalgae as a sustainable energy source for biodiesel production: a review. Renewable and Sustainable Energy Reviews 15, 584–593.
Alves, L. P. S., Almeida, A.T., Cruz, L.M., Pedrosa, F.O., de Souza, E.M., Cubatsu, L.S., Müller-Santos, M. and Valdameri, G., 2017, A simple and efficient method for poly-3-hydroxybutyrate quantification in diazotrophic bacteria within 5 minutes using flow cytometry, Brazilian Journal of Medical and Biological Research 50(1), 1–10.
Aransiola, E. F., Ojumu, T. V., Oyekola, O. O., Madzimbamuto, T. F. & Ikhu–Omoregbe, D. I. O., 2014, A review of current technology for biodiesel production: state of the art, Biomass and Bioenergy 61, 276–297.
Balat, M., 2011, Potential alternatives to edible oils for biodiesel production–a review of current work, Energy Conversion and Management 52(2), 1479–1492.
Behrens, P. W. & Kyle, D. J., 1996, Microalgae as a source of fatty acids, Journal of Food Lipids 3, 259–272.
Bradford, M. M., 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Analytical Biochemistry 72, 248–254.
Chen, W., Zhang, C., Song, L., Sommerfeld, M. and Hu, Q., 2009, A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae, Journal of Microbiological Methods 77, 41–47.
Chen, J., Li, J., Dong, W., Zhang, X., Tyagi, R. D., Drogul, P. and Surampalli, R. Y., 2018, The potential of microalgae in biodiesel production, Renewable and Sustainable Energy Reviews 90, 336–346.
Chisti, Y., 2007, Biodiesel from microalgae, Biotechnology Advances 25, 294–306.
Courchesne, N. M. D., Parisien, A., Wang, B. and Lan, C. Q., 2009, Enhancement of lipid production using biochemical, genetic and transcription factor engineering approaches, Journal of Biotechnology 14(1), 31–41.
Davis, M. S., Solbiati, J. and Cronan, Jr. J. E., 2000, Overproduction of acetyl-CoA carboxylase activity increases the rate of fatty acid biosynthesis in Escherichia coli, Journal of Biological Chemistry 275(37), 28593–28598.
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. & Smith, F., 1956, Colorimetric method for determination of sugars and related substances, Analytical Chemistry 28(3), 350–356.
Fan, J., Cui, Y., Wan, M., Wang, W. & Li, Y., 2014, Lipid accumulation and biosynthesis genes response of the oleaginous Chlorella pyrenoidosa under three nutrition stressors, Biotechnology for Biofuels 7(17).
Fang, M., Jin, L., Zhang, C., Tan, Y., Jiang, P., Ge, N., Li, H. & Xing, X., 2013, Rapid mutation of Spirulina platensis by a new mutagenesis system of atmospheric and room temperature plasmas (ARTP) and generation of a mutant library with diverse phenotypes, PLoS ONE 8(10), 1–12.
Faried, M., Samer, M., Abdelsalam, E., Yousef, R. S., Attia, Y. A. & Ali, A. S., 2017, Biodiesel production from microalgae: Processes, technologies and recent advancements, Renewable and Sustainable Energy Reviews 79, 893–913.
Feng, Y., Li C. & Zhang, D., 2011, Lipid production of Chlorella vulgaris cultured in artificial wastewater medium, Bioresource Technology 102(1), 969–975.
Goh, B. H. H., Ong, H. C., Cheah, M. Y., Chen, W., Yu, K. L. & Mahlia, T. M. I., 2019, Sustainability of direct biodiesel synthesis from microalgae biomass: A critical review, Renewable and Sustainable Energy Reviews 107, 59–74.
Halim, R. & Webley, P. A., 2015, Nile Red Staining for Oil Determination in Microalgal Cells: A New Insight through Statistical Modelling, International Journal of Chemical Engineering.
Huang, G. H., Chen, G. & Chen, F., 2009, Rapid screening method for lipid production in alga based on Nile red fluorescence, Biomass and Bioenergy 33(10), 1386–1392.
Hosseini Tafreshi, A. & Shariari, M., 2009, Dunaliella biotechnology: methods and applications, Journal of Applied Microbiology 107(1), 14–35.
Kim, K. H., 1997, Regulation of mammalian acetyl-coenzyme A carboxylase, Annual Review of Nutrition, 77–99.
Knothe, G., 2009, Improving biodiesel fuel properties by modifying fatty ester composition, Energy and Environmental Science 2(7), 759–766.
Knothe, G., 2010, Biodiesel and renewable diesel: A comparison, Progress in Energy and Combustion Science 36(3), 364–373.
Li, S. J. & Cronan, Jr. J. E., 1993, Growth rate regulation of Escherichia coli acetyl coenzyme A carboxylase, which catalyzes the first committed step of lipid biosynthesis, Journal of Bacteriology 175(2), 332–340.
Liu, S., Zhao, Y., Liu, L., Ao, X., Ma, L., Wu, M. & Ma, F., 2015, Improving cell growth and lipid accumulation in green microalgae Chlorella sp. via UV irradiation, Applied Biochemistry and Biotechnology 175, 3507–3518.
Nascimento, I. A., Marquez, S.S.I., Cabanelas, I.T.D., Pereira, S.A., Druzian, J.I., de Souza, C.O., Vich, D.V., de Carvalho, G.C. & Nascimento, M.A., 2013, Screening microalgae strains for biodiesel production: Lipid productivity and estimation of fuel quality based on fatty acids profiles as selective criteria, Bioenergy Research 6(1), 1–13.
Rahman D Y., Rachmayati R., Widyaningrum D. N. & Susilaningsih D., 2020, Enhancement of lipid production of Chlorella sp. 042 by mutagenesis, IOP Conference Series: Earth and Environmental Sciences (EES) 439, 1-6.
Rawat, I., Ranjith Kumar, R., Mutanda, T. & Bux, F., 2013, Biodiesel from microalgae: A critical evaluation from laboratory to large scale production, Applied Energy 103, 444–467.
Rostron, K. A., Rolph, C. E. & Lawrence, C. L., 2015, Nile red fluorescence screening facilitating neutral lipid phenotype determination in budding yeast, Saccharomyces cerevisiae, and the fission yeast Schizosaccharomyces pombe, Antonie van Leeuwenhoek, International Journal of General and Molecular Microbiology 108(1), 97–106.
Ryckebosch, E., Muylaert, K. & Foubert, I., 2012, Optimization of an analytical procedure for extraction of lipids from microalgae, Journal of the American Oil Chemists’s Socíecty’ 89, 189–198.
Sarayloo, E., Tardu, M., Unlu, Y. S., Simsek, S., Cehavir, G., Erkey, C. & Kavakli, I. H., 2017, Understanding lipid metabolism in high–lipid–producing Chlorella vulgaris mutants at the genome–wide level, Algal Research 28, 244–252.
Sarayloo, E., Simsek, S., Unlu, Y. S., Cevahir, G., Erkey, C. & Kavakli, I. H., 2018, Enhancement of the lipid productivity and fatty acid methyl ester profile of Chlorella vulgaris by two rounds of mutagenesis, Bioresource Technology 250, 764–769.
Satpati, G. G. & Pal, R., 2014, Rapid detection of neutral lipid in green microalgae by flow cytometry in combination with Nile red staining an improved technique, Annals of Microbiology 65(2), 937–949.
Sendl, A., Schliack, M., Loser, R., Stanislaus, F. & Wagner, H., 1992, Inhibition of cholesterol synthesis in vitro by extracts and isolated compounds prepared from garlic and wild garlic, Atherosclerosis 94(1), 79–85.
Sharma, K., Li, Y. & Schenk, P. M., 2014, UV-C-mediated lipid induction and settling, a step change towards economical microalgal biodiesel production, Green Chemistry, 16(7), 3539–3548.
Sitepu, I. R., Jin, M., Fernandez, J.E., da Costa Sousa, L., Balan, V. & Boundy-Mills, K.L., 2014, Identification of oleaginous yeast strains able to accumulate high intracellular lipids when cultivated in alkaline pretreated corn stover, Applied Microbiology and Biotechnology 98(17), 7645–7657.
Sivaramakrishnan, R. & Incharoensakdi, A., 2017, Enhancement of lipid production in Scenedesmus sp. by UV mutagenesis and hydrogen peroxide treatment, Bioresource Technology 235, 366–370.
Tan, X., Uemura, Y., Lim, J. W., Wong, C. Y. & Lee, K. T., 2017, Cultivation of microalgae for biodiesel production: A review on upstream and downstream processing, Chinese Journal of Chemical Engineering 26(1), 17–30.
Tapia, E. V., Anschau, A., Coradini, A.LV., Franco, T.T. & 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), 1–8.
Trentacoste, E. M., Shrestha, R.P., Smith, S.R., Gle, C., Hartmann, A.C., Hildebrand, M. & Gerwick, W.H., 2013, Metabolic engineering of lipid catabolism increases microalgal lipid accumulation without compromising growth, Proceedings of the National Academy of Sciences of the United States of America 110(49), 19748–19753.
Van Vooren, G., Le Grand, F., Legrand, J., Cuine, S., Peltier, G. & Pruvost, J., 2012, Investigation of fatty acids accumulation in Nannochloropsis oculata for biodiesel application, Bioresource Technology 124, 421–432.
Vigeolas, H., Duby, F., Kaymak, E., Niessen, G., Motte, P., Franck, F. & Remacle, C., 2012, Isolation and partial characterization of mutants with elevated lipid content in Chlorella sorokiniana and Scenedesmus obliquus, Journal of Biotechnology 162, 3–12.
Yoo C., Jun S. Y., Lee J. Y. Ahn C. Y. & Oh H. M., 2010, Selection of microalgae for lipid production under high levels carbon dioxide, Bioresource Technology, 101(Suppl 1), pp. S71–S74.
Yusuf, N. N. A. N., Kamarudin, S. K. & Yaakub, Z., 2010, Overview on the current trends in biodiesel production, Energy Conversion and Management 52, 2741–2751.
Zhang, X. Z., Hu, Q., Sommerfeld, M., Puruhito, E. & Chen, Y.S., 2010, Harvesting algal biomass for biofuels using ultrafiltration membranes, Bioresource Technology 101, 5297–5304.
DOI: https://doi.org/10.22146/jtbb.56862
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