Microalgae Flocculation

Biotechnology Advances 20 (2003) 491 – 515 www.elsevier.com/locate/biotechadv Recovery of microalgal biomass and metabolites: process options and economics
´ ´ E. Molina Grimaa, E.-H. Belarbia, F.G. Acien Fernandeza, a b,* A. Robles Medina , Yusuf Chisti b ´ ´ Department of Chemical Engineering, University of Almerıa, E-04071 Almerıa, Spain Institute of Technology and Engineering PN456, Massey University, Private Bag 11 222, Palmerston North, New Zealand

a

Received 27 May 2002; received in revised form 15 October 2002; accepted 15 October 2002

Abstract Commercial production of intracellular microalgal metabolites requires the following: (1) largescale monoseptic production of the appropriate microalgal biomass; (2) recovery of the biomass from a relatively dilute broth; (3) extraction of the metabolite from the biomass; and (4) purification of the crude extract. This review examines the options available for recovery of the biomass and the intracellular metabolites from the biomass. Economics of monoseptic production of microalgae in photobioreactors and the downstream recovery of metabolites are discussed using eicosapentaenoic acid (EPA) recovery as a representative case study. D 2002 Elsevier Science Inc. All rights reserved.
Keywords: Microalgae; Phaeodactylum tricornutum; Eicosapentaenoic acid; Process economics; Flocculation; Biomass recovery

1. Introduction Microalgae can be used to produce numerous high-value bioactives (Borowitzka, 1986; Bubrick, 1991; Pulz et al., 2001; Li et al., 2001; Banerjee et al., 2002). Production of
* Corresponding author. Tel.: [phone]; fax: [phone]. E-mail address: [e-mail] (Y. Chisti). 0734-9750/02/$ – see front matter D 2002 Elsevier Science Inc. All rights reserved. PII: S 0 7 3 4 - 9 7 5 0 ( 0 2 ) 0 0 0 5 0 - 2

492

E. Molina Grima et al. / Biotechnology Advances 20 (2003) 491–515

microalgae-derived metabolites requires processes for culturing the alga (Ben-Amotz and

References: ´ ´ ´ ´ ´ Acien Fernandez FG, Garcıa F, Sanchez JA, Fernandez JM, Molina E. Modelling of biomass productivity in tubular photobioreactors for microalgal cultures: effects of dilution rate, tube diameter and solar irradiance. Biotechnol Bioeng 1998;58:605 – 16. ´ ´ ´ ´ ´ Acien Fernandez FG, Fernandez Sevilla JM, Sanchez Perez JA, Molina Grima E, Chisti Y. Airlift-driven external-loop tubular photobioreactors for outdoor production of microalgae: assessment of design and performance. Chem Eng Sci 2001;56: 2721 – 32. Banerjee A, Sharma R, Chisti Y, Banerjee UC. Botryococcus braunii: a renewable source of hydrocarbons and other chemicals. Crit Rev Biotechnol 2002;22:245 – 79. Belarbi H, Molina E, Chisti Y. A process for high and scaleable recovery of high purity eicosapentaenoic acid esters from microalgae and fish oil. Enzyme Microb Technol 2000;26:516 – 29. Belter PA, Cussler EL, Hu WS. Bioseparations: Downstream Processing for Biotechnology. New York: Wiley; 1988. Ben-Amotz A, Avron M. The biotechnology of mass culturing of Dunaliella for products of commercial interest. In: Cresswell RC, Rees TAV, Shah N, editors. Algal and cyanobacterial technology. London: Longman; 1987. p. 90 – 114. Benemann JR, Kopman BL, Weissman DE, Eisenberg DE, Goebel RP. Development of microalgae harvesting and high rate pond technologies in California. In: Shelef G, Soeder CJ, editors. Algal biomass. Amsterdam: Elsevier; 1980. p. 457. ´ Bermejo Roman R, Talavera EM, Alvarez-Pez JM. Chromatographic purification and characterization of b-phycoerythrin from Porphyridium cruentum. Semipreparative HPLC separation and characterization of its subunits. J Chromatogr A 2001;917:135 – 45. ´ ´ ´ ´ Bermejo Roman R, Alvarez-Pez JM, Acien Fernandez FG, Molina Grima E. Recovery of pure b-phycoerythrin from the microalga Porphyridium cruentum. J Biotechnol 2002;93:73 – 85. Bilanovic D, Shelef G, Sukenik A. Flocculation of microalgae with cationic polymers—effects of medium salinity. Biomass 1988;17:65 – 76. E. Molina Grima et al. / Biotechnology Advances 20 (2003) 491–515 513 Blanchemain A, Grizeau D. Increased production of eicosapentaenoic acid by Skeletonema costatum cells after decantation at low temperature. Biotechnol Tech 1999;13:497 – 501. Borowitzka MA. Microalgae as sources of fine chemicals. Microbiol Sci 1986;3:372 – 5. Borowitzka MA. Closed algal photobioreactors: design considerations for large-scale systems. J Mar Biotechnol 1996;4: 185 – 91. Borowitzka MA. Microalgae for aquaculture: opportunities and constraints. J Appl Phycol 1997;9:393 – 401. Borowitzka MA. Commercial production of microalgae: ponds, tanks, tubes and fermenters. J Biotechnol 1999;70:313 – 21. Bubrick P. Production of astaxanthin from Haematococcus. Biores Technol 1991;38:237 – 9. Cannell RJP. Algal biotechnology. Appl Biochem Biotechnol 1990;26:85 – 105. ´ ´˜ Cartens M, Molina E, Robles A, Gimenez A, Ibanez MJ. Eicosapentaenoic acid (20:4n-3) from the marine microalga Phaeodactylum tricornutum. J Am Oil Chem Soc 1996;73:1025 – 31. Chisti Y. Strategies in downstream processing. In: Subramanian G, editor. Bioseparation and bioprocessing: a handbook vol. 2. New York: Wiley-VCH; 1998. p. 3 – 30. Chisti Y. Shear sensitivity. In: Flickinger MC, Drew SW, editors. Encyclopedia of bioprocess technology: fermentation, biocatalysis, and bioseparation, vol. 5. New York: Wiley; 1999. p. 2379 – 406. Chisti Y, Moo-Young M. Disruption of microbial cells for intracellular products. Enzyme Microb Technol 1986;8:194 – 204. Chisti Y, Moo-Young M. Fermentation technology, bioprocessing, scale-up and manufacture. In: Moses M, Cape RE, editors. Biotechnology: the science and the business. New York: Harwood Academic Publishers; 1991. p. 167 – 209. Cohen JM, Rourke GA, Woodward RL. Natural and synthetic polyelectrolytes as coagulants and coagulant aids. Bull U S Dept Health, Educ Welf [Robert A Taft Sanitary Eng Center, Cinncinati, OH, December]. Dodd JC. Algae production and harvesting from animal wastewaters. Agric Wastes 1979;1:23 – 37. D’Souza FML, Lecossois D, Heasman MP, Diemar JA, Jackson CJ, Pendrey RC. Evaluation of centrifuged microalgae concentrates as diets for Penaeus monodon Fabricius larvae. Aquacult Res 2000;31:661 – 70. Dunstan GA, Vokman JK, Jeffrey SW, Barret SM. Biochemical composition of microalgae from the green algal classes Chlorophyceae and Parasinophyceae: 2. Lipid classes and fatty acid. J Exp Mar Biol Ecol 1992;161:115 – 34. Elmaleh S, Coma J, Grasmick A, Bourgade L. Magnesium induced algal flocculation in a fluidized bed. Water Sci Technol 1991;23:1695 – 702. ´ ´ ´˜ ´ Gimenez Gimenez A, Ibanez MJ, Robles A, Molina E, Garcıa S, Esteban L. Downstream processing and purification of eicosapentaenoic (20:5n-3) and arachidonic acids (20:4n-6) from the microalga Porphyridium cruentum. Bioseparation 1998;7:89 – 99. Golueke CG, Oswald WJ. Harvesting and processing sewage grown planktonic algae. J Water Pollut Control Fed 1965;37: 471 – 98. Gudin C, Chaumont D. Cell fragility, the key problem of microalgae mass production in closed photobioreactors. Bioresour Technol 1991;38:141 – 51. Gudin C, Therpenier C. Bioconversion of solar energy into organic chemicals by microalgae. Adv Biotechnol Processes 1986;6:73 – 110. Heasman M, Diemar J, O’ Connor W, Sushames T, Foulkes L, Nell JA. Development of extended shelf-life microalgae concentrate diets harvested by centrifugation for bivalve molluscs—a summary. Aquacult Res 2000;31(8 – 9):637 – 59. ´ Hedenskog G, Enebo L, Vendlova J, Prokes BTI. Investigation of some methods for increasing the digestibility in vitro of microalgae. Biotechnol Bioeng 1969;11:37 – 51. Hee-Mock O, Lee SJ, Park MH, Kim HS, Kim HC, Yoon JH, et al. Harvesting of Chlorella vulgaris using a bioflocculant from Paenibacillus sp. AM49. Biotechnol Lett 2001;23:1229 – 34. Humphreys K. Jelen’s cost and optimization engineering. 3rd ed. New York: McGraw-Hill; 1991. ´˜ ´ ´ ´ Ibanez Gonzalez MJ, Robles Medina A, Molina Grima E, Gimenez Gimenez A, Carstens M, Esteban Cerdan L. Optimization of fatty acid extraction from Phaeodactylum tricornutum UTEX 640 biomass. J Am Oil Chem Soc 1998;75:1735 – 40. Jiang J.-Q., Graham NJD, Harward C. Comparison of polyferric sulphate with other coagulants for the removal of algae and algae-derived organic matter. Water Sci Technol 1993;27:221 – 30. Kaya VM, Picard G. Stability of chitosan gel as entrapment matrix of viable Scenedesmus bicellularis cells immobilized on screens for tertiary treatment of wastewater. Bioresour Technol 1996;56:147 – 55. Koopman B, Lincoln EP. Autoflotation harvesting of algae from high rate pond effluents. Agric Wastes 1983;5:231 – 46. Lavoie A, de la Noue J. Harvesting microalgae with chitosan. J World Maricult Soc 1983;14:685 – 94. Lee SJ, Kim SB, Kim JE, Kwon GS, Yoon BD, Oh HM. Effects of harvesting method and growth stage on the flocculation of the green alga Botryococcus braunii. Lett Appl Microbiol 1998;27:14 – 8. Li HB, Chen F, Zhang TY, Yang FQ, Xu GQ. Preparative isolation and purification of lutein from the microalga Chlorella vulgaris by high-speed counter-current chromatography. J Chromatogr A 2001;905:151 – 5. 514 E. Molina Grima et al. / Biotechnology Advances 20 (2003) 491–515 Lim GB, Lee SY, Lee EK, Haam SJ, Kim WS. Separation of astaxanthin from red yeast Phaffia rhodozyma by supercritical carbon dioxide extraction. Biochem Eng J 2002;11:181 – 7. Lincoln EP. Resource recovery with microalgae. Arch Hydrobiol 1985;20:25 – 34. Lubian LM. Concentrating cultured marine microalgae with chitosan. Aquacult Eng 1989;8:257 – 65. MacKay D, Salusbury T. Choosing between centrifugation and crossflow microfiltration. Chem Eng (Lond) 1988;477: 45 – 50. McCausland MA, Brown MR, Barrett SM, Diemar JA, Heasman MP. Evaluation of live microalgae and microbial pastes as supplementary food for juvenile Pacific oyster (Crassostrea gigas). Aquaculture 1999;174:323 – 42. McGarry MG. Alga flocculation with aluminium sulfate and polyelectrolytes. J Water Pollut Control Fed 1970;42:R191 – 201. Mendes-Pinto MM, Raposo MFJ, Bowen J, Young AJ, Morais R. Evaluation of different cell disruption processes on encysted cells of Haematococcus pluvialis: effects on astaxanthin recovery and implications for bio-availability. J Appl Phycol 2001;13:19 – 24. Middelberg APJ. The release of intracellular bioproducts. In: Subramanian G, editor. Bioseparation and bioprocessing: a handbook, vol. 2. Weinheim: Wiley-VCH; 1994. p. 131 – 64. Mohn FH. Improved technologies for the harvesting and processing of microalgae and their impact on production costs. Arch Hydrobiol, Beih Ergeb Limnol 1978;1:228 – 53. Mohn FH. Experiences and strategies in the recovery of biomass from mass cultures of microalgae. In: Shelef G, Soeder CJ, editors. Algae biomass. Amsterdam: Elsevier; 1980. p. 547 – 71. Molina Grima E. Microalgae, mass culture methods. In: Flickinger MC, Drew SW, editors. Encyclopedia of bioprocess technology: fermentation, biocatalysis and bioseparation, vol. 3. New York: Wiley; 1999. p. 1753 – 69. ´ ´ ´ ´ ´ ´ ´ Molina Grima E, Robles Medina A, Gimenez Gimenez A, Sanchez Perez JA, Garcıa Camacho F, Garcıa Sanchez JL. Comparison between extraction of lipids and fatty acids from microalgal biomass. J Am Oil Chem Soc 1994;71:955 – 9. ´ ´ ´ ˜ Molina Grima E, Robles Medina A, Gimenez Gimenez A, Ibanez Gonzalez MJ. Gram-scale purification of eicosapentaenoic acid (EPA 20:5n3) from wet Phaeodactylum tricornutum UTEX 640 biomass. J Appl Phycol 1996;8:359 – 67. ´ ´ Molina Grima E, Acien Fernandez FG, Garcia Camacho F, Chisti Y. Photobioreactors: light regime, mass transfer, and scale-up. J Biotechnol 1999;70:231 – 48. Moraine R, Shelef G, Sandbank F, Bar-Moshe Z, Shvartzbard I. Recovery of sewage borne algae: flocculation and centrifugation technique. In: Shelef G, Soeder CJ, editors. Algae biomass. Amsterdam: Elsevier; 1980. p. 531 – 46. Morales J, de la Noue J, Picard G. Harvesting marine microalgae species by chitosan flocculation. Aquaculture Eng 1985;4: 257 – 70. Nigam BP, Ramanathan PK, Venkataraman LV. Application of chitosan as a flocculant for the cultures of the green alga: Scenedesmus acutus. Arch Hydrobiol 1980;88:378 – 87. Peet M, Brind J, Ramchand CN, Shah S, Vankar GK. Two double-blind placebo-controlled pilot studies of eicosapentaenoic acid in the treatment of schizophrenia. Schizophr Res 2001;49:243 – 51. Peet M, Horrobin DFEPA Multicentre Study Group. A dose-ranging exploratory study of the effects of ethyl-eicosapentaenoate in patients with persistent schizophrenic symptoms. J Psychiatr Res 2002;36:7 – 18. Peters MS, Timmerhaus KD. Plant design and economics for chemical engineers. 4th ed. New York: McGraw-Hill; 1991. Petrusevski B, Bolier G, Van Bremen AN, Alaerts GJ. Tangential flow filtration: a method to concentrate freshwater algae. Water Res 1995;29:1419 – 24. Pulz O. Photobioreactors: production systems for phototrophic microorganisms. Appl Microbiol Biotechnol 2001;57:287 – 93. Pulz O, Scheinbenbogen K. Photobioreactors: design and performance with respect to light energy input. Adv Biochem Eng Biotechnol 1998;59:123 – 52. Pulz O, Scheibenbogen K, Gross W. Biotechnology with cyanobacteria and microalgae. In: Rehm H.-J., Reed G, editors. Biotechnology, vol. 10. Weinheim: Wiley-VCH; 2001. p. 105 – 36. Pushparaj B, Pelosi E, Torzillo G, Materassi R. Microbial biomass recovery using a synthetic cationic polymer. Bioresour Technol 1993;43:59 – 62. Richmond A. Microalgae of economic potential. In: Richmond A, editor. CRC handbook of microalgal mass culture. Boca Raton: CRC Press; 1986. p. 199 – 243. Richmond A, Becker EW. Technological aspects of mass cultivation, a general outline. In: Richmond A, editor. CRC handbook of microalgal mass culture. Boca Raton: CRC Press; 1986. p. 245 – 63. ´ ´ ´ ´ ´ ´ Robles Medina A, Gimenez Gimenez A, Garcıa Camacho F, Sanchez Perez JA, Molina Grima E, Contreras Gomez A. Concentration and purification of stearidonic, eicosapentaenoic, and docosahexaenoic acids from cod liver oil and the marine microalga Isochrysis galbana. J Am Oil Chem Soc 1995;72:575 – 83. ´ ´ ´ ´ Rodrıguez-Ruiz J, Belarbi E.-H., Garcıa Sanchez JL, Lopez Alonso D. Rapid simultaneous lipids extraction and transesterification for fatty acid analysis. Biotechnol Tech 1998;12:689 – 91. E. Molina Grima et al. / Biotechnology Advances 20 (2003) 491–515 515 Rossignol N, Vandanjon L, Jaouen P, Quemeneur F. Membrane technology for the continuous separation microalgae/culture medium: compared performances of cross-flow microfiltration and ultrafiltration. Aquacult Eng 1999;20:191 – 208. Ruane, M. Extraction of caroteniferous materials from algae. Australian patent No. 7,239,574 (1977). ´ ´ ´ ´ Sanchez Miron A, Contreras Gomez A, Garcıa Camacho F, Molina Grima E, Chisti Y. Comparative evaluation of compact photobioreactors for large-scale monoculture of microalgae. J Biotechnol 1999;70:249 – 70. Shelef G. Photosynthetic biomass production from sewage. Arch Hydrobiol Beih 1978;11:3 – 14. Suh HH, Kwon GS, Lee CH, Kim HS, Oh HM, Yoon BD. Characterization of bioflocculant produced by Bacillus sp. DP-152. J Ferment Bioeng 1997;84:108 – 12. Tenney MW, Echelberger WF, Schuessler RG, Pavoni JL. Algal flocculation with synthetic organic polyelectrolytes. Appl Bacteriol 1969;18:965 – 71. Terry KL, Raymond LP. System design for the autotrophic production of microalgae. Enzyme Microb Technol 1985;7:474 – 87. Tilton RC, Murphy J, Dixon JK. The flocculation of algae with synthetic polymeric flocculants. Water Res 1972;6:155 – 64. Tredici MR. Bioreactors, photo. In: Flickinger MC, Drew SW, editors. Encyclopedia of bioprocess technology: fermentation, biocatalysis and bioseparation, vol. 1. New York: Wiley; 1999. p. 395 – 419. Venkataraman LV. New possibility for microalgae production and utilisation in India. Arch Hydrobiol Beih 1978;11:199 – 210. Yahi H, Elmaleh S, Coma J. Algal flocculation – sedimentation by pH increase in a continuous reactor. Water Sci Technol 1994;30:259 – 67.