Abstract
In this study the thermo-resistant green alga Micractinium sp. ME05 was cultivated in media containing molasses as a carbon source. Shake flask experiments and 2-L bioreactor experiments were conducted at different inoculum ratios, aeration rates, and agitation speeds. The experimental condition which resulted in the highest biomass concentration (3.73 ± 0.45 g L−1) with 10% inoculum in 500-mL flasks was scaled up to 2-L flasks at two aeration rates (0.25 and 0.5 L min−1). An increase in biomass concentration from 2.35 ± 0.53 to 3.06 ± 0.21 g L−1 was observed with an increase of aeration rate from 0.25 to 0.50 L min−1, which demonstrated significant effect of aeration rate on biomass concentration (p = 0.000 < 0.05). In 2-L bioreactor experiments, highest biomass productivity (0.53 ± 0.076 g L−1 day−1) and lipid productivity (7.7 ± 1.6 g L−1 day−1) were obtained with 5% (v/v) inoculum and 50 rpm agitation speed. The principal fatty acids were palmitic acid (C16:0) and linoleic acid (C18:2) comprising 30.2 ± 1.01 and 45.2 ± 1.32% of the total fatty acid content, respectively. Thus, the present study highlights the possibility of using molasses for biomass and lipid production with Micractinium sp. ME05 under different cultivation conditions. Using low cost feedstock such as molasses would be valuable in terms of evaluating waste materials for further biodiesel production.
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References
Abou-Shanab RA, Raghavulu SV, Hassanin NMA, Kim s KYJ, Oh SU, Jeon B-H (2012) Manipulating nutrient composition of microalgal growth media to improve biomass yield and lipid content of Micractinium pusillum. Afr J Biotechnol 11:16270–16276
Azma M, Mohamed MS, Mohamad R, Rahmin RA, Ariff AB (2011) Improvement of medium composition for heterotrophic cultivation of green microalgae, Tetraselmis suecica, using response surface methodology. Biochem Eng J 53:187–195
Barbosa MJ, Albrecht M, Wijffels RH (2003) Hydrodynamic stress and lethal events in sparged microalgae cultures. Biotechnol Bioeng 83:112–120
Borowitzka MA (2013) High-value products from microalgae—their development and commercialisation. J Appl Phycol 25:743–756
Borowitzka MA, Moheimani NR (2013) Open pond culture systems. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 133–152
Borowitzka MA, Vonshak A (2017) Scaling up microalgal cultures to commercial scale. Eur J Phycol 52:407–418
Bouarab L, Dauta A, Loudiki M (2004) Heterotrophic and mixotrophic growth of Micractinium pusillum Fresenius in the presence of acetate and glucose: effect of light and acetate gradient concentration. Water Res 38:2706–2712
Carvalho AP, Meireles LA, Malcata FX (2006) Microalgal reactors: a review of enclosed system designs and performances. Biotechnol Prog 22:1490–1506
Chen F (1996) High cell density culture of microalgae in heterotrophic growth. Trends Biotechnol 14:421–426
Chen G-Q, Chen F (2006) Growing phototrophic cells without light. Biotechnol Lett 28:607–616
Cheng Y, Lu Y, Gao C, Wu Q (2009a) Alga-based biodiesel production and optimization using sugar cane as the feedstock. Energy Fuel 23:4166–4173
Cheng Y, Zhou W, Gao C, Lan K, Gao Y, Wu Q (2009b) Biodiesel production from Jerusalem artichoke (Helianthus Tuberosus L. ) tuber by heterotrophic microalgae Chlorella protothecoides. J Chem Technol Biotechnol 84:777–781
Chisti Y (2007) Biodiesel from microalgae. Biotechnol Adv 25:294–306
Chiu SY, Kao CY, Chen CH, Kuan TC, Ong SC, Lin CS (2008) Reduction of CO2 by a high-density culture of Chlorella sp. in a semicontinuous photobioreactor. Bioresour Technol 99:3389–3396
Contin A, Van Der Heijden R, Ten Hoopen HJG, Verpoorte R (1998) The inoculum size triggers tryptamine or secologanin biosynthesis in a Catharanthus roseus cell culture. Plant Sci 139:205–211
De Swaaf ME, Sijtsma L, Pronk JT (2003) High-cell-density fed-batch cultivation of the docosahexaenoic acid producing marine alga Crypthecodinium cohnii. Biotechnol Bioeng 81:666–672
Miller GL (1959) Use of dinitrosalicyclic acid reagent for determination of reducing sugar. Anal Chem 31:426–428
Gao C, Zhai Y, Ding Y, Wu Q (2010) Application of sweet sorghum for biodiesel production by heterotrophic microalga Chlorella protothecoides. Appl Energy 87:756–761
Gaurav K, Srivastava R, Sharma JG, Singh R, Singh V (2015) Molasses based growth and lipid production by Chlorella pyrenoidosa: a potential feedstock for biodiesel. Int J Green Energy 13:320–327
Gorman DS, Levine RP (1965) Cytochrome f and plastocyanin: their sequence in the photosynthetic electron transport chain of Chlamydomonas reinhardi. Proc Natl Acad Sci U S A 54:1665–1669
Graverholt OS, Eriksen NT (2007) Heterotrophic high-cell-density fed-batch and continuous-flow cultures of Galdieria sulphuraria and production of phycocyanin. Appl Microbiol Biotechnol 77:69–75
Griffiths MJ, Harrison STL (2009) Lipid productivity as a key characteristic for choosing algal species for biodiesel production. J Appl Phycol 21:493–507
Gurkok S, Cekmecelioglu D, Ogel ZB (2011) Optimization of culture conditions for Aspergillus sojae expressing an Aspergillus fumigatus α-galactosidase. Bioresour Technol 102:4925–4929
Heidari M, Kariminia HR, Shayegan J (2016) Effect of culture age and initial inoculum size on lipid accumulation and productivity in a hybrid cultivation system of Chlorella vulgaris. Process Saf Environ Prot 104:111–122
Heredia-Arroyo T, Wei W, Hu B (2010) Oil accumulation via heterotrophic/mixotrophic Chlorella protothecoides. Appl Biochem Biotechnol 162:1978–1995
Huang G, Chen F, Wei D, Zhang X, Chen G (2010) Biodiesel production by microalgal biotechnology. Appl Energy 87:38–46
Huang J, Li Y, Wan M, Yan Y, Feng F, Qu X, Wang J, Shen G, Li W, Fan J, Wang W (2014) Novel flat-plate photobioreactors for microalgae cultivation with special mixers to promote mixing along the light gradient. Bioresour Technol 159:8–16
Ji MK, Abou-Shanab RAI, Kim SH, Salama E, Lee SH, Kabra AN, Lee Y-S, Hong S, Jeon B-H (2013) Cultivation of microalgae species in tertiary municipal wastewater supplemented with CO2 for nutrient removal and biomass production. Ecol Eng 58:142–148
Karpagam R, Raj KJ, Ashokkumar B, Varalakshmi P (2015) Characterization and fatty acid profiling in two fresh water microalgae for biodiesel production: lipid enhancement methods and media optimization using response surface methodology. Bioresour Technol 188:177–184
Knothe G (2009) Improving biodiesel fuel properties by modifying fatty ester composition. Energy Environ Sci 2:759–766
Knothe G (2013) Production and properties of biodiesel from algal oils. In: Borowitzka MA, Moheimani NR (eds) Algae for biofuels and energy. Springer, Dordrecht, pp 207–221
Kose Engin I, Cekmecelioglu D, Yücel AM, Oktem HA (2018) Enhancement of heterotrophic biomass production by Micractinium sp.ME05. Waste Biomass Valorization 9:811–820
Kumar V, Muthuraj M, Palabhanvi B, Ghoshal AK, Das D (2014) High cell density lipid rich cultivation of a novel microalgal isolate Chlorella sorokiniana FC6 IITG in a single-stage fed-batch mode under mixotrophic condition. Bioresour Technol 170:115–124
Liang Y, Sarkany N, Cui Y (2009) Biomass and lipid productivities of Chlorella vulgaris under autotrophic, heterotrophic and mixotrophic growth conditions. Biotechnol Lett 31:1043–1049
Lin Z, Raya A, Ju LK (2014) Microalga Ochromonas danica fermentation and lipid production from waste organics such as ketchup. Process Biochem 49:1383–1392
Liu J, Huang J, Jiang Y, Chen F (2012) Molasses-based growth and production of oil and astaxanthin by Chlorella zofingiensis. Bioresour Technol 107:393–398
Liu J, Sun Z, Zhong Y, Gerken H, Huang J, Chen F (2013) Utilization of cane molasses towards cost-saving astaxanthin production by a Chlorella zofingiensis mutant. J Appl Phycol 25:1447–1456
Lu L, Wang J, Yang G, Zhu B, Pan K (2017) Heterotrophic growth and nutrient productivities of Tetraselmis chuii using glucose as a carbon source under different C/N ratios. J Appl Phycol 29:15–21
Ma Q, Wang J, Lu S, Lv Y, Yuan Y (2013) Quantitative proteomic profiling reveals photosynthesis responsible for inoculum size dependent variation in Chlorella sorokiniana. Biotechnol Bioeng 110:773–784
Moheimani NR, Isdepsky A, Lisec J, Raes E, Borowitzka MA (2011) Coccolithophorid algae culture in closed photobioreactors. Biotechnol Bioeng 108:2078–2087
Morales-Sánchez D, Tinoco-Valencia R, Kyndt J, Martinez A (2013) Heterotrophic growth of Neochloris oleoabundans using glucose as a carbon source. Biotechnol Biofuels 6:100
Najafpour GD, Poi Shan C (2003) Enzymatic hydrolysis of molasses. Bioresour Technol 86:91–94
Onay M, Sonmez C, Oktem HA, Yucel AM (2014) Thermo-resistant green microalgae for effective biodiesel production: isolation and characterization of unialgal species from geothermal flora of central Anatolia. Bioresour Technol 169:62–71
Onay M, Sonmez C, Oktem HA, Yucel M (2016) Evaluation of various extraction techniques for efficient lipid recovery from thermo-resistant microalgae, Hindakia, Scenedesmus and Micractinium species. Am J Anal Chem 7:141–150
Pahl SL, Lewis DM, Chen F, King KD (2010) Growth dynamics and the proximate biochemical composition and fatty acid profile of the heterotrophically grown diatom Cyclotella cryptica. J Appl Phycol 22:165–171
Park KC, Whitney C, McNichol JC, Dickinson KE, MacQuarrie S, Skrupski BP, Zou J, Wilson KE, O'Leary JB, McGinn PJ (2012) Mixotrophic and photoautotrophic cultivation of 14 microalgae isolates from Saskatchewan, Canada: potential applications for wastewater remediation for biofuel production. J Appl Phycol 24:339–348
Perez-Garcia O, Escalante FME, de-Bashan LE, Bashan Y (2011) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res 45:11–36
Piasecka A, Krzemińska I, Tys J (2017) Enrichment of Parachlorella kessleri biomass with bioproducts: oil and protein by utilization of beet molasses. J Appl Phycol 29:1735–1743
Posten C (2009) Design principles of photo-bioreactors for cultivation of microalgae. Eng Life Sci 9:165–177
Anderson RA (2005) Photobioreactors and fermentors: the light and dark sides of growing algae. In: Andersen Robert (ed) Algal culturing techniques. Elsevier Academic Press, NY pp 189–203
Singhasuwan S, Choorit W, Sirisansaneeyakul S, Kokkaew N, Chisti Y (2015) Carbon-to-nitrogen ratio affects the biomass composition and the fatty acid profile of heterotrophically grown Chlorella sp. TISTR 8990 for biodiesel production. J Biotechnol 216:169–177
Smith RT, Bangert K, Wilkinson SJ, Gilmour DJ (2015) Synergistic carbon metabolism in a fast growing mixotrophic freshwater microalgal species Micractinium inermum. Biomass Bioenergy 82:73–86
Sobczuk TM, Camacho FG, Grima EM, Chisti Y (2006) Effects of agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. Bioprocess Biosyst Eng 28:243–250
Sonmez C, Elcin E, Akin D, Oktem HA, Yucel M (2016) Evaluation of novel thermo-resistant Micractinium and Scenedesmus sp. for efficient biomass and lipid production under different temperature and nutrient regimes. Bioresour Technol 211:422–428
Stephenson AL, Dennis JS, Howe CJ, Scott SA, Smith AG (2010) Influence of nitrogen-limitation regime on the production of Chlorella vulgaris of lipids for biodiesel feedstocks. Biofuels 1:47–58
Wei A, Zhang X, Wei D, Chen G, Wu Q, Yang ST (2009) Effects of cassava starch hydrolysate on cell growth and lipid accumulation of the heterotrophic microalgae Chlorella protothecoides. J Ind Microbiol Biotechnol 36:1383–1389
Yan D, Lu Y, Chen YF, Wu Q (2011) Waste molasses alone displaces glucose-based medium for microalgal fermentation towards cost-saving biodiesel production. Bioresour Technol 102:6487–6493
Yeh KL, Chang JS (2012) Effects of cultivation conditions and media composition on cell growth and lipid productivity of indigenous microalga Chlorella vulgaris ESP-31. Bioresour Technol 105:120–127
Yen HW, Chang JT (2015) Growth of oleaginous Rhodotorula glutinis in an internal-loop airlift bioreactor by using lignocellulosic biomass hydrolysate as the carbon source. J Biosci Bioeng 119:580–584
Acknowledgements
This study was carried out at the Middle East Technical University (METU) Biology Department Plant Biotechnology Laboratory and METU Food Engineering Department Bioprocess Laboratory. We would like to thank Asst. Prof. Dr. Melih Onay for his isolation and characterization of microalgal species used in this study.
Funding
This study was funded by the Scientific and Technological Research Council of Turkey (TUBITAK) Project Number 114Z487).
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Engin, I.K., Cekmecelioglu, D., Yücel, A.M. et al. Heterotrophic growth and oil production from Micractinium sp. ME05 using molasses. J Appl Phycol 30, 3483–3492 (2018). https://doi.org/10.1007/s10811-018-1486-2
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DOI: https://doi.org/10.1007/s10811-018-1486-2