Abstract
Many studies have demonstrated that heterotrophic and mixotrophic growth for various microalgae species yields greater biomass and lipid content as compared to photoautotrophic cultivation. This review explores the possibility of leveraging the natural ability of the microorganisms to metabolize carbon heterotrophically and mixotrophically in agricultural wastewaters. This has the potential advantage of improving the overall economics for the production of biodiesel and value-added biomolecules from microalgae, mitigating an existing waste stream and minimizing water requirements. However, there are a number of challenges and gaps in scientific knowledge that suggest a need for ongoing research in the area. In this review, specific focus is dedicated to the metabolic mechanisms, reported performances, and practical challenges that contribute to the uncertainty of employing agricultural wastewaters for heterotrophic and mixotrophic microalgae cultures.
Similar content being viewed by others
References
Ahn JH, Do TH, Kim SD, Hwang S (2006) The effect of calcium on the anaerobic digestion treating swine wastewater. Biochem Eng J 30:33–38
Amer L, Adhikari B, Pellegrino J (2011) Technoeconomic analysis of five microalgae-to-biofuels processes of varying complexity. Bioresour Technol 102:9350–9359
Anderson RA (ed) (2005) Algae Culturing Techniques. Elsevier Academic Press, NY
Appels L, Baeyens J, Degrève J, Dewil R (2008) Principles and potential of the anaerobic digestion of waste-activated sludge. Prog Energ Combust 34:755–781
Arbeli Z, Brenner A, Abeliovich A (2006) Treatment of high-strength dairy wastewater in an anaerobic deep reservoir: analysis of the methanogenic fermentation pathway and the rate-limiting step. Water Res 40:3653–3659
Barsanti L, Gualtieri P (2006) Algae: anatomy, biochemistry, and biotechnology. CRC Press, Boca Raton
Ben W, Qiang Z, Adams C, Zhang H, Chen L (2008) Simultaneous determination of sulfonamides, tetracyclines and tiamulin in swine wastewater by solid-phase extraction and liquid chromatography-mass spectrometry. J Chromatogr 1202:173–180
Bhatnagar A, Chinnasamy S, Singh M, Das KC (2011) Renewable biomass production by mixotrophic algae in the presence of various carbon sources and wastewaters. Appl Energ 88:3425–3431
Bohutskyi P, Kula T, Kessler BA, Hong Y, Bouwer EJ, Betenbaugh MJ, Allnutt FCT (2014) Mixed trophic state production process for microalgal biomass with high lipid content for generating biodiesel and biogas. Bioenerg Res 1–12
Borowitzka MA (1999) Commercial production of microalgae: ponds, tanks, tubes and fermenters. J Biotechnol 70:313–321
Borowitzka MA (2013) High-value products from microalgae—their development and commercialisation. J Appl Phycol 25:743–756
Brennan L, Owende P (2010) Biofuels from microalgae—a review of technologies for production, processing, and extractions of biofuels and co-products. Ren Sust Energ Rev 14:557–577
Bumbak F, Cook S, Zachleder V, Hauser S, Kovar K (2011) Best practices in heterotrophic high-cell-density microalgal processes: achievements, potential and possible limitations. Appl Microbiol Biotechnol 91:31–46
Burkholder JM, Glibert PM, Skelton HM (2008) Mixotrophy, a major mode of nutrition for harmful algal species in eutrophic waters. Harmful Algae 8:77–93
Campbell PK, Beer T, Batten D (2011) Life cycle assessment of biodiesel production from microalgae in ponds. Bioresour Technol 102:50–56
Cañizares-Villanueva RO, Domínguez AR, Cruz MS, Ríos-Leal E (1995) Chemical composition of cyanobacteria grown in diluted, aerated swine wastewater. Bioresour Technol 51:111–116
Chang KJL, Rye L, Dunstan GA, Grant T, Koutoulis A, Nichols PD, Blackburn SI (2014) Life cycle assessment: Heterotrophic cultivation of thraustochytrids for biodiesel production. J Appl Phycol. doi:10.1007/s10811-014-0364-9
Chen F (1996) High cell density culture of microalgae in heterotrophic growth. Trends Biotech 14:421–426
Chen F, Johns MR (1996) Heterotrophic growth of Chlamydomonas reinhardtii on acetate in chemostat culture. Process Biochem 31:601–604
Chen T, Wang Y (2013) Optimized astaxanthin production in Chlorella zofingiensis under dark condition by response surface methodology. Food Sci Biotechnol 1–8
Chen F, Zhang Y (1997) High cell density mixotrophic culture of Spirulina platensis on glucose for phycocyanin production using a fed-batch system. Enzyme Microb Technol 20:221–224
Chen F, Chen H, Gong X (1997) Mixotrophic and heterotrophic growth of Haematococcus lacustris and rheological behaviour of the cell suspensions. Bioresour Technol 62:19–24
Chen C-Y, Yeh K-L, Aisyah R, Lee DJ, Chang JS (2011) Cultivation, photobioreactor design and harvesting of microalgae for biodiesel production: a critical review. Bioresour Technol 102:71–81
Chisti Y (2007) Biodiesel from microalgae. Biotehnol Adv 25:294–306
Chisti Y (2013) Constraints to commercialization of algal fuels. J Biotechnol 167:201–214
Cole KM, Sheath RG (eds) (1990) Biology of the Red Algae. Cambridge University Press, Cambridge
De Bashan LE, Bashan Y (2004) Recent advances in removing phosphorus from wastewater and its future use as fertilizer (1997–2003). Water Res 38:4222–4246
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
Del Campo J, García-González M, Guerrero M (2007) Outdoor cultivation of microalgae for carotenoid production: current state and perspectives. Appl Microbiol Biot 74:1163–1174
Droop MR (1974) Heterotrophy of Carbon. In: Stewart WDP (ed) Algal Physiology and Biochemistry. Blackwell Scientific, Oxford, pp 530–559
Fang X, Wei C, Zhao-Ling C, Fan O (2004) Effects of organic carbon sources on cell growth and eicosapentaenoic acid content of Nannochloropsis sp. J Appl Phycol 16:499–503
Feng Y, Li C, Zhang D (2011) Lipid production of Chlorella vulgaris cultured in artificial wastewater medium. Bioresour Technol 102:101–105
Ferguson JL (2002) Characterizing the process of composting mink manure and pelted mink carcasses. Master’s Thesis, Nova Scotia Agricultural College, Truro, Canada
Fu R, Wang H, Pei G (2012) A study on lipid production of the mixotrophic microalgae Phaeodactylum tricornutum on various carbon sources. Afr J Microbiol Res 6:1041–1047
Gao C, Zhai Y, Ding Y, Wu Q (2010) Application of sweet sorghum for biodiesel production by heterotrophic microalga Chlorella protothecoides. Appl Energ 87:756–761
García-Fernández JM, De Marsac NT, Diez J (2004) Streamlined regulation and gene loss as adaptive mechanisms in Prochlorococcus for optimized nitrogen utilization in oligotrophic environments. Microbiol Mol Biol Rev 68:630–638
Ghaly AE, Kamal M, Mahmoud NS (2005) Phytoremediation of aquaculture wastewater for water recycling and production of fish feed. Environ Int 31:1–13
González C, Marciniak J, Villaverde S et al (2008) Microalgae-based processes for the biodegradation of pretreated piggery wastewaters. Appl Microbiol Biotechnol 80:891–898
Guillen-Jimenez E, Alvarez-Mateos P, Romero-Guzman F, Pereda-Marin J (2000) Bio-mineralization of organic matter in dairy wastewater, as affected by pH. The evolution of ammonium and phosphates. Water Res 34:1215–1224
Hammouda O, Gaber A, Abdelraouf N (1995) Microalgae and wastewater treatment. Ecotox Env Safety 31:205–210
Hunt PG, Szogi AA, Humenik FJ et al (2002) Constructed wetlands for the treatment of swine wastewater from an anaerobic lagoon. Trans ASAE 45:639–647
Ip PF, Chen F (2005) Production of astaxanthin by the green microalga Chlorella zofingiensis in the dark. Process Biochem 40:733–738
Ip P-F, Wong K-H, Chen F (2004) Enhanced production of astaxanthin by the green microalga Chlorella zofingiensis in mixotrophic culture. Process Biochem 39:1761–1766
Kim MK, Choi K-M, Yin C-R, Lee K-Y, Im W-T, Lim JH, Lee S-T (2004) Odorous swine wastewater treatment by purple non-sulfur bacteria, Rhodopseudomonas palustris, isolated from eutrophicated ponds. Biotechnol Lett 26:819–822
Kim MK, Park JW, Park CS, Kim SJ, Jeune KH, Chang MU, Acreman J (2007) Enhanced production of Scenedesmus spp. (green microalgae) using a new medium containing fermented swine wastewater. Bioresour Technol 98:2220–2228
Kong Q, Li L, Martinez B, Chen P, Ruan R (2010) Culture of microalgae Chlamydomonas reinhardtii in wastewater for biomass feedstock production. Appl Biochem Biotechnol 160:9–18
Lee RE (1980) Phycology, 1st edn. Press, Cambridge University
Lee Y-K (2001) Microalgal mass culture systems and methods: their limitation and potential. J Appl Phycol 13:307–315
Lee YK, Ding SY, Hoe CH, Low CS (1996) Mixotrophic growth of Chlorella sorokiniana in outdoor enclosed photobioreactor. J Appl Phycol 8:163–169
Leyva LA, Bashan Y, Mendoza A, de Bashan LE (2014) Accumulation fatty acids of in Chlorella vulgaris under heterotrophic conditions in relation to activity of acetyl-CoA carboxylase, temperature, and co-immobilization with Azospirillum brasilense. Naturwissenschaften 101:819–830
Li X, Xu H, Wu Q (2007) Large‐scale biodiesel production from microalga Chlorella protothecoides through heterotrophic cultivation in bioreactors. Biotechnol Bioeng 98:764–771
Li Y, Horsman M, Wu N, Lan CQ, Dubois-Calero N (2008) Biofuels from microalgae. Biotechnol Prog 24:815–820
Li P, Miao X, Li R, Zhong J (2011) In situ biodiesel production from fast-growing and high oil content Chlorella pyrenoidosa in rice straw hydrolysate. J Biomed Biotechnol 2011:1–8
Li Y, Zhou W, Hu B, Min M, Chen P, Ruan RR (2012) Effect of light intensity on algal biomass accumulation and biodiesel production for mixotrophic strains Chlorella kessleri and Chlorella protothecoide cultivated in highly concentrated municipal wastewater. Biotechnol Bioeng 109:2222–2229
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 Y-F, Jing S-R, Lee D-Y, Wang T-W (2002) Nutrient removal from aquaculture wastewater using a constructed wetlands system. Aquaculture 209:169–184
Lowrey J, Yildiz I (2011) Seawater/wastewater production of microalgae-based biofuels in closed loop tubular photobioreactors. Master Thesis California Polytechnic State University, San Luis Obispo, USA, 141 pp
Lowrey J, Yildiz I (2014) Investigation of heterotrophic cultivation potential of Chlorella vulgaris and Tetraselmis chuii in controlled environment wastewater growth media from dairy, poultry and aquaculture industries. Acta Horticult 1037:1109–1114
Marquez FJ (1995) Enhancement of biomass and pigment production during growth of Spirulina platensis in mixotrophic culture. J Chem Technol Biot 62:159–164
Mata TM, Martins AA, Caetano NS (2010) Microalgae for biodiesel production and other applications: a review. Renew Sust Energ Rev 14:217–232
McGinn PJ, Dickinson KE, Bhatti S, Frigon JC, Guiot SR, O’Leary SJ (2011) Integration of microalgae cultivation with industrial waste remediation for biofuel and bioenergy production: opportunities and limitations. Photosynth Res 109:231–247
McGinn PJ, Dickinson KE, Park KC, Whitney CG, MacQuarrie SP, Black FJ, Frigon J-C, Guiot SR, O’Leary SJB (2012) Assessment of the bioenergy and bioremediation potentials of the microalga Scenedesmus sp. AMDD cultivated in municipal wastewater effluent in batch and continuous mode. Algal Res 1:155–165
Mendes A, Reis A, Vasconcelos R, Guerra P, Lopes da Silva T (2009) Crypthecodinium cohnii with emphasis on DHA production: a review. J Appl Phycol 21:199–214
Miao X, Wu Q (2006) Biodiesel production from heterotrophic microalgal oil. Bioresour Technol 97:841–846
Morris I (1974) Nitrogen assimilation and protein synthesis. In: Stewart WDP (ed) Algal Physiology and Biochemistry. Blackwell Scientific Publs, Oxford, pp 583–609
Mulbry W, Kondrad S, Buyer J, Luthria DL (2009) Optimization of an oil extraction process for algae from the treatment of manure effluent. J Am Oil Chem Soc 86:909–915
Obaja D, Macé S, Costa J, Sans C, Mata-Alvarez J (2003) Nitrification, denitrification and biological phosphorus removal in piggery wastewater using a sequencing batch reactor. Bioresour Technol 87:103–111
Ogawa T, Aiba S (1981) Bioenergetic analysis of mixotrophic growth in Chlorella vulgaris and Scenedesmus acutus. Biotechnol Bioeng 23:1121–1132
Ogbonna JC, Tanaka H (1998) Cyclic autotrophic/heterotrophic cultivation of photosynthetic cells: a method of achieving continuous cell growth under light/dark cycles. Bioresour Technol 65:65–72
Ogbonna JC, Tanaka H (2000) Light requirement and photosynthetic cell cultivation—development of processes for efficient light utilization in photobioreactors. J Appl Phycol 12:207–218
Ogbonna JC, Yoshizawa H, Tanaka H (2000) Treatment of high strength organic wastewater by a mixed culture of photosynthetic microorganisms. J Appl Phycol 12:277–284
Okubo Y, Futamata H, Hiraishi A (2006) Characterization of phototrophic purple nonsulfur bacteria forming colored microbial mats in a swine wastewater ditch. Appl Environ Microbiol 72:6225–6233
Olguín EJ (2003) Phycoremediation: key issues for cost-effective nutrient removal processes. Biotechnol Adv 22:81–91
Passell H, Dhaliwal H, Reno M, Wu B, Ben Amotz A, Ivry E, Gay M, Czartoski T, Laurin L, Ayer N (2013) Algae biodiesel life cycle assessment using current commercial data. J Environ Manag 129:103–111
Pate R, Klise G, Wu B (2011) Resource demand implications for US algae biofuels production scale-up. Appl Energ 88:3377–3388
Paul JW, Beauchamp EG (1989) Effect of carbon constituents in manure on denitrification in soil. Can J Soil Sci 69:49–61
Perez-Garcia O, De-Bashan LE, Hernandez J-P, Bashan Y (2010) Efficiency of growth and nutrient uptake from wastewater by heterotrophic, autotrophic, and mixotrophic cultivation of Chlorella vulgaris immobilized with Azospirillum brasilense. J Phycol 46:800–812
Perez-Garcia O, Bashan Y, Esther Puente M (2011a) Organic carbon supplementation of sterilized municipal wastewater is essential for heterotrophic growth and removing ammonium by the microalga Chlorella vulgaris. J Phycol 47:190–199
Perez-Garcia O, Escalante FME, de Bashan LE, Bashan Y (2011b) Heterotrophic cultures of microalgae: metabolism and potential products. Water Res 45:11–36
Prathima Devi M, Venkata Subhash G, Venkata Mohan S (2012) Heterotrophic cultivation of mixed microalgae for lipid accumulation and wastewater treatment during sequential growth and starvation phases: effect of nutrient supplementation. Renew Energ 43:276–283
Queiroz MI, Hornes MO, da Silva G, Manetti A, Zepka LQ, Jacob-Lopes E (2013) Fish processing wastewater as a platform of the microalgal biorefineries. Biosyst Eng 115:195–202
Ratledge C, Kanagachandran K, Anderson AJ, Grantham D, Stephenson J (2001) Production of docosahexaenoic acid by Crypthecodinium cohnii grown in a pH-auxostat culture with acetic acid as principal carbon source. Lipids 36:1241–1246
Rawat I, Ranjith Kumar R, Mutanda T, Bux F (2011) Dual role of microalgae: phycoremediation of domestic wastewater and biomass production for sustainable biofuels production. Appl Energ 88:3411–3424
Redfield AC (1934) On the proportions of organic derivatives in sea water and their relation to the composition of plankton. In: Daniel RJ (ed) James Johnstone Memorial Volume. Liverpool University Press, Liverpool, pp 176–192
Roessler PG (1988) Changes in the activities of various lipid and carbohydrate biosynthetic enzymes in the diatom Cyclotella cryptica in response to silicon deficiency. Arch Biochem Biophys 267:521–528
Salminen EA, Rintala JA (2002) Semi-continuous anaerobic digestion of solid poultry slaughterhouse waste: effect of hydraulic retention time and loading. Water Res 36:3175–3182
Sánchez S, Martínez ME, Espejo MT, Pacheco R, Espinola F, Hodaifa G (2001) Mixotrophic culture of Chlorella pyrenoidosa with olive-mill wastewater as the nutrient medium. J Appl Phycol 13:443–449
Severin BF (1980) Disinfection of municipal wastewater effluents with ultraviolet light. J Water Pollut Contr Fedn 52:2007–2018
Sijtsma L, Anderson AJ, Ratledge C (2005) Alternative carbon sources for heterotrophic production of docosahexaenoic acid by the marine alga Crypthecodinium cohnii. In: Ratledge C, Cohen Z (eds) Single Cell Oils. AOCS Press, Champaign, pp 107–123
Singh A, Nigam PS, Murphy JD (2011) Mechanism and challenges in commercialisation of algal biofuels. Bioresour Technol 102:26–34
Sloth JK, Wiebe MG, Eriksen NT (2006) Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria. Enzyme Microb Tech 38:168–175
Sooknah RD, Wilkie AC (2004) Nutrient removal by floating aquatic macrophytes cultured in anaerobically digested flushed dairy manure wastewater. Ecol Eng 22:27–42
Sun A, Davis R, Starbuck M, Ben-Amotz A, Pate R, Pienkos P (2011) Comparative cost analysis of algal oil production for biofuels. Energy 36:5169–5179
Syrett PJ, Morris I (1963) The inhibition of nitrate assimilation by ammonium in Chlorella. Biochim Biophys Acta 67:566–575
Tabernero A, Martín del Valle EM, Galán MA (2012) Evaluating the industrial potential of biodiesel from a microalgae heterotrophic culture: scale-up and economics. Biochem Eng J 63:104–115
Tam NF, Wong Y (2000) Effect of immobilized microalgal bead concentrations on wastewater nutrient removal. Environ Pollut 107:145–151
Tanoi T, Kawachi M, Watanabe MM (2010) Effects of carbon source on growth and morphology of Botryococcus braunii. J Appl Phycol 23:25–33
Tett P, Droop MR, Heaney SI (1985) The Redfield Ratio and phytoplankton growth rate. J Mar Biol Assoc UK 65:487–504
Tripathi U, Sarada R, Ravishankar GA (2002) Effect of culture conditions on growth of green alga—Haematococcus pluvialis and astaxanthin production. Acta Physiol Plant 24:323–329
Wan M, Liu P, Xia J, Rosenberg JN, Oyler GA, Betenbaugh MJ, Nie Z, Qiu G (2011) The effect of mixotrophy on microalgal growth, lipid content, and expression levels of three pathway genes in Chlorella sorokiniana. Appl Microbiol Biotechnol 91:835–844
Wang L, Min M, Li Y, Chen Y, Liu Y, Wang Y, Ruan R (2009) Cultivation of green algae Chlorella sp. in different wastewaters from municipal wastewater treatment plant. Appl Biochem Biotechnol 162:1174–1186
Wang L, Li Y, Chen P, Min M, Chen Y, Zhu J, Ruan RR (2010) Anaerobic digested dairy manure as a nutrient supplement for cultivation of oil-rich green microalgae Chlorella sp. Bioresour Technol 101:2623–2628
Wang H, Xiong H, Hui Z, Zeng X (2012) Mixotrophic cultivation of Chlorella pyrenoidosa with diluted primary piggery wastewater to produce lipids. Bioresour Technol 104:215–220
Wang J, Yang H, Wang F (2014) Mixotrophic cultivation of microalgae for biodiesel production: status and prospects. Appl Biochem Biotechnol 172:3307–3329
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
Weyer KM, Bush DR, Darzins A, Willson BD (2010) Theoretical maximum algal oil production. Bioenerg Res 3:204–213
Wigmosta MS, Coleman AM, Skaggs RJ, Huesemann MH, Lane LJ (2011) National microalgae biofuel production potential and resource demand. Water Resour Res 47: W00H04.
Wilkie AC, Mulbry WW (2002) Recovery of dairy manure nutrients by benthic freshwater algae. Bioresour Technol 84:81–91
Williams PJB L, Laurens LML (2010) Microalgae as biodiesel & biomass feedstocks: Review & analysis of the biochemistry, energetics & economics. Energ Environ Sci 3:554–590
Xu H, Miao X, Wu Q (2006) High quality biodiesel production from a microalga Chlorella protothecoides by heterotrophic growth in fermenters. J Biotechnol 126:499–507
Yamane Y, Utsunomiya T, Watanabe M, Sasaki K (2001) Biomass production in mixotrophic culture of Euglena gracilis under acidic condition and its growth energetics. Biotechnol Lett 23:1223–1228
Yang C, Hua Q, Shimizu K (2000) Energetics and carbon metabolism during growth of microalgal cells under photoautotrophic, mixotrophic and cyclic light-autotrophic/dark-heterotrophic conditions. Biochem Eng J 6:87–102
Yang C, Ding Z, Zhang K (2008) Growth of Chlorella pyrenoidosa in wastewater from cassava ethanol fermentation. World J Microbiol Biotechnol 24:2919–2925
Yetilmezsoy K, Sakar S (2008) Improvement of COD and color removal from UASB treated poultry manure wastewater using Fenton’s oxidation. J Hazard Mater 151:547–558
Yetilmezsoy K, Sapci-Zengin Z (2009) Recovery of ammonium nitrogen from the effluent of UASB treating poultry manure wastewater by MAP precipitation as a slow release fertilizer. J Hazard Mater 166:260–269
Zhang H, Wang W, Li Y, Yang W, Shen G (2011) Mixotrophic cultivation of Botryococcus braunii. Biomass Bioenergy 35:1710–1715
Zhou W, Min M, Li Y, Hu B, Ma X, Cheng Y, Liu Y, Chen P, Ruan R (2012) A hetero-photoautotrophic two-stage cultivation process to improve wastewater nutrient removal and enhance algal lipid accumulation. Bioresour Technol 110:448–455
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lowrey, J., Brooks, M.S. & McGinn, P.J. Heterotrophic and mixotrophic cultivation of microalgae for biodiesel production in agricultural wastewaters and associated challenges—a critical review. J Appl Phycol 27, 1485–1498 (2015). https://doi.org/10.1007/s10811-014-0459-3
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-014-0459-3