Abstract
The freshwater microalga Chlorella vulgaris was grown heterotrophically in fed-batch 50–600-L fermenters at 36°C, on aerated and mixed nutrient solution with urea as a nitrogen and glucose as a carbon and energy source. Cell density increased from the initial value 6.25 to 117.18 g DW L−1 in 32 h in the fermenter 50 L at a mean growth rate 3.52 g DW L−1 h−1. The DW increase in the fermenter 200 L was from 7.25 to 94.82 g DW L−1 in 26.5 h at a mean growth rate 3.37 g DW L−1 h−1. Mean specific growth rate μ was about 0.1 h−1 in the both fermenters, if nutrients and oxygen were adequately supplied. The DW increase in the fermenter 600 L was from 0.8 to 81.6 g DW L−1 in 66.5 h at a mean growth rate 1.22 g DW L−1 h−1 and μ = 0.07 h−1. A limitation of the cell growth rate in 600 L fermenter caused by a low dissolved oxygen concentration above cell densities higher than 10 g DW L−1) occurred. Specific growth rate decreased approximately linearly with increasing glucose concentration (25–80 g glucose L−1) at the beginning of cultivation and decreased with the time of cultivation. The cell yield was 0.55–0.69 g DW (g glucose)−1. The content of proteins, β-carotene, and chlorophylls in the cells steadily increased and starch content decreased, by keeping aerated and mixed culture another 12 h in fermenter after the cell growth was stopped due to glucose deficiency.
Similar content being viewed by others
References
Borowitzka MA (1992) Algal biotechnology products and processes-matching science and economics. J Appl Phycol 4:267–279
Borowitzka MA (1999) Commercial production of microalgae: ponds, tanks, tubes and fermentors. J Biotechnol 70:313–321
Chen F (1996) High cell density culture of microalgae in heterotrophic growth. Trends Biotechnol 14:421–426
Chen GQ, Chen F (2006) Growing phototrophic cells without light. Biotechnol Lett 28:607–616
Doucha J, Lívanský K (2001) Method of controlled cultivation of algae in heterotrophic mode of nutrition. Czech Patent 288638
Doucha J, Lívanský K (2006) Productivity, CO2/O2 exchange and hydraulics in outdoor open high density microalgal (Chlorella sp.) photobioreactors operated in Middle and Southern European climate. J Appl Phycol 18:811–826
Doucha J, Lívanský K (2008) Chlorella vulgaris BEIJ. strain Doucha et Lívanský 1996/H 14. Czech Patent 299352, 2008
Doucha J, Lívanský K, Kotrbáček V, Zachleder V (2009) Production of Chlorella biomass enriched by selenium and its use in animal nutrition: a review. Appl Microbiol Biotechnol 83:1001–1008
El-Sheekh MM, Fathy AA (2009) Variation of some nutritional constituents and fatty acids profiles of Chlorella vulgaris Beijerinck grown under auto and heterotrophic conditions. Internat J Botany 5:153–159
Endo H, Shirota M (1972) Studies on the heterotrophic growth of Chlorella in a mass culture. In: Proc IV IFS: Ferment Technol Today 533–541
Endo H, Nakajima K, Chino R, Shirota M (1974) Growth characteristics and cellular composition of Chlorella regularis, heterotrophic fast growing strain. Agr Biol Chem 38:9–18
Eriksen NT (2008) The technology of microalgal culturing. Biotechnol Lett 30:1525–1536
Harel M, Place AR (2004) Heterotrophic production of marine algae for aquaculture. In: Richmond A (ed) Handbook of microalgal culture: biotechnology and applied phycology. Blackwell, New York, pp 513–524
Lee YK (1997) Commercial production of microalgae in the Asia-Pacific rim. J Appl Phycol 9:403–411
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
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
Lowry OH, Rosenbrough NS, Farr AL, Randall RJ (1951) Protein measurement with the folin-phenol reagent. J Biol Chem 193:265–275
Materassi R, Paoletti C, Balloni W, Florenzano G (1980) Some considerations on the production of lipid substances by microalgae and cyanobacteria. In: Shelef G, Soeder CJ (eds) Algae biomass production and use. Elsevier, Amsterdam, pp 619–626
McCready RM, Guggolz J, Silviera V, Owens HS (1950) Determination of starch and amylose in vegetables. Anal Chem 22:1156–1158
Pulz O (2001) Photobioreactors: production systems for phototrophic microorganisms. Appl Microbiol Biotechnol 57:287–293
Sansawa H, Endo H (2004) Production of intracellular phytochemicals in Chlorella under heterotrophic conditions. J Biosci Bioeng 98:437–444
Shi XM, Chen F (2008) High-yield production of lutein by the green microalga Chlorella protothecoides in heterotrophic fed-batch culture. Biotechnol Prog 18:723–728
Shi XM, Chen F, Yuan JP, Chen H (1997) Heterotrophic production of lutein by selected Chlorella strains. J Appl Phycol 9:445–450
Shi XM, Zhang XW, Chen F (2000) Heterotrophic production of biomass and lutein by Chlorella protothecoides on various nitrogen sources. Enzyme Microb Technol 27:312–318
Sun N, Wang Y, Li YT, Huang JC, Chen F (2008) Sugar-based growth, astaxanthin accumulation and carotenogenic transcription of heterophic Chlorella zofingiensis (Chlorophyta). Process Biochem 43:1288–1292
Tredici MR (2004) Mass production of microalgae: photobioreactors. In: Richmond A (ed) Handbook of microalgal culture. Blackwell, Oxford, pp 178–214
Wu Z, Shi X (2007) Optimization for high-density cultivation of heterotrophic Chlorella based on a hybrid neural network model. Lett Appl Microbiol 44:13–18
Wu ZY, Shi CL, Shi XM (2007) Modeling of lutein production by heterotrophic Chlorella in batch and fed-batch cultures. World J Microbiol Biotechnol 23:1233–1238
Xiong W, Li X, Xiang J, Wu Q (2008) High-density fermentation of microalga Chlorella protothecoides in bioreactor for microbio-diesel production. Appl Microbiol Biotechnol 78:29–36
Zbíral J (2009) Unified working procedures for testing of feeds: determination of β-carotene content by HPLC method. (Norm 240, in Czech). ÚKZÚZ, Prague. p. 1–6
Acknowledgments
This work was partially supported by the EUREKA project Alganol OE 09025 of the Ministry of Education, Youth and Sports of the Czech Republic. The authors thank for the technical cooperation to the team of the Center of Biotechnologies, Institute of Microbiology, Acad. Sci. of the Czech Republic, Prague, where part of the growth experiments were realized.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Doucha, J., Lívanský, K. Production of high-density Chlorella culture grown in fermenters. J Appl Phycol 24, 35–43 (2012). https://doi.org/10.1007/s10811-010-9643-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10811-010-9643-2