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Long-term storage of microalgae: determination of optimum cryopreservation conditions

Published online by Cambridge University Press:  01 August 2022

Irem Deniz*
Affiliation:
Bioengineering Department, Faculty of Engineering, Manisa Celal Bayar University, 45119, Yunusemre/Manisa, Turkey
Zeliha Demirel
Affiliation:
Bioengineering Department, Faculty of Engineering, Ege University, 35100, Bornova/Izmir, Turkey
Esra Imamoglu
Affiliation:
Bioengineering Department, Faculty of Engineering, Ege University, 35100, Bornova/Izmir, Turkey
Meltem Conk-Dalay
Affiliation:
Bioengineering Department, Faculty of Engineering, Ege University, 35100, Bornova/Izmir, Turkey
*
Author for correspondence: Irem Deniz, E-mail: [email protected]

Abstract

Maintenance of eukaryotic microalgae strains for the long term is generally carried out using serial subculture techniques which require labour, time and cost. Cryopreservation techniques provide long-term storage of up to years for numerous microorganism strains and cell cultures. Ssu930ijn vbvbhnn8;l,n is related to a successfully designed mass and heat transfer balance throughout the cell. In this study, optimization of the cryopreservation process was carried out for two commercially used microalgal strains. The parameters to be optimized were DMSO percentage (0–25%), incubation time (1–15 min) and cryopreservation term (7–180 days) using a central composite design (CCD). Long-term storage up to 123.17 and 111.44 days corresponding to high cell viabilities was achieved for Chlorella vulgaris and Neochloris texensis, respectively. Generated models were found to be in good agreement with experimental results. The study also revealed holistic results for storage of microalgal strains in a stable state for industrial applications.

Type
Research Article
Copyright
Copyright © The Author(s), 2022. Published by Cambridge University Press on behalf of Marine Biological Association of the United Kingdom

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References

Ammar, EE, Aioub, AA, Elesawy, AE, Karkour, AM, Mouhamed, MS, Amer, AA and El-Shershaby, NA (2022) Algae as bo-fertilizers: between current situation and future prospective. Saudi Journal of Biological Sciences 29, 30833096.Google ScholarPubMed
Apt, KE and Behrens, PW (1999) Commercial developments in microalgal biotechnology. Journal of Phycology 35, 215226.Google Scholar
Bezerra, MA, Santelli, RE, Oliveira, EP, Villar, LS and Escaleira, LA (2008) Response surface methodology (RSM) as a tool for optimization in analytical chemistry. Talanta 76, 965977.CrossRefGoogle ScholarPubMed
Bligh, EG and Dyer, WJ (1959) A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology 37, 911917.CrossRefGoogle ScholarPubMed
Bradford, MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72, 248254.Google ScholarPubMed
Bui, TV, Ross, IL, Jakob, G and Hankamer, B (2013) Impact of procedural steps and cryopreservation agents in the cryopreservation of chlorophyte microalgae. PLoS ONE 8, e78668.CrossRefGoogle ScholarPubMed
Day, JG (2007) Cryopreservation of microalgae and cyanobacteria. In Cryopreservation and Freeze-Drying Protocols. New York, NY: Springer, pp. 141151.CrossRefGoogle Scholar
Day, J and Fleck, R (2015) Cryo-injury and the implications this has to the conservation of biological resources. Microalgae Biotechnology 1, 111.CrossRefGoogle Scholar
Day, JG and Stacey, G (2007) Cryopreservation and Freeze-Drying Protocols. New York, NY: Springer Science & Business Media.CrossRefGoogle Scholar
Day, J, Watanabe, M, Morris, GJ, Fleck, R and McLellan, M (1997) Long-term viability of preserved eukaryotic algae. Journal of Applied Phycology 9, 121127.CrossRefGoogle Scholar
Day, J, Benson, EE, Harding, K, Knowles, B, Idowu, M, Bremner, D, Santos, F, Friedl, T, Lorenz, M, Lukesova, A, Elster, J, Lukavsky, J, Herdman, M, Rippka, R and Hall, T (2005) Cryopreservation and conservation of microalgae: the development of a pan-European scientific and biotechnological resource (the COBRA project). CryoLetters 26, 231238.Google Scholar
Ernst, A, Deicher, M, Herman, PM and Wollenzien, UI (2005) Nitrate and phosphate affect cultivability of cyanobacteria from environments with low nutrient levels. Applied and Environmental Microbiology 71, 33793383.CrossRefGoogle ScholarPubMed
Fernandes, MS, Calsing, LC, Nascimento, RC, Santana, H, Morais, PB, de Capdeville, G and Brasil, BS (2019) Customized cryopreservation protocols for chlorophytes based on cell morphology. Algal Research 38, 101402.CrossRefGoogle Scholar
Gaget, V, Chiu, Y-T, Lau, M and Humpage, AR (2017) From an environmental sample to a long-lasting culture: the steps to better isolate and preserve cyanobacterial strains. Journal of Applied Phycology 29, 309321.CrossRefGoogle Scholar
Grima, EM, Pérez, JS, Camacho, FG, Fernández, FA, Alonso, DL and Del Castillo, CS (1994) Preservation of the marine microalga, Isochrysis galbana: influence on the fatty acid profile. Aquaculture 123, 377385.CrossRefGoogle Scholar
Guermazi, W, Sellami-Kammoun, A, Elloumi, J, Drira, Z, Aleya, L, Marangoni, R, Ayadi, H and Maalej, S (2010) Microalgal cryo-preservation using dimethyl sulfoxide (Me2SO) coupled with two freezing protocols: influence on the fatty acid profile. Journal of Thermal Biology 35, 175181.CrossRefGoogle Scholar
Guler, BA, Deniz, I, Demirel, Z, Oncel, SS and Imamoglu, E (2020) Computational fluid dynamics modelling of stirred tank photobioreactor for Haematococcus pluvialis production: hydrodynamics and mixing conditions. Algal Research 47, 101854.CrossRefGoogle Scholar
Harding, K (2010) Plant and algal cryopreservation: issues in genetic integrity, concepts in cryobionomics and current applications in cryobiology. Asia-Pacific Journal of Molecular Biology and Biotechnology 18, 151154.Google Scholar
Imamoglu, E, Demirel, Z and Conk Dalay, M (2015) Process optimization and modeling for the cultivation of Nannochloropsis sp. and Tetraselmis striata via response surface methodology. Journal of Phycology 51, 442453.CrossRefGoogle ScholarPubMed
Isleten-Hosoglu, M, Ayyıldız-Tamis, D, Zengin, G and Elibol, M (2013) Enhanced growth and lipid accumulation by a new Ettlia texensis isolate under optimized photoheterotrophic condition. Bioresource Technology 131, 258265.Google ScholarPubMed
Kapoore, RV, Huete-Ortega, M, Day, JG, Okurowska, K, Slocombe, SP, Stanley, MS and Vaidyanathan, S (2019) Effects of cryopreservation on viability and functional stability of an industrially relevant alga. Scientific Reports 9, 112.CrossRefGoogle ScholarPubMed
Kim, M, Kim, D, Cho, JM, Nam, K, Lee, H, Nayak, M, Han, J-I, Oh, H-M and Chang, YK (2021) Hydrodynamic cavitation for bacterial disinfection and medium recycling for sustainable Ettlia sp. cultivation. Journal of Environmental Chemical Engineering 9, 105411.Google Scholar
Konar, N, Durmaz, Y, Genc Polat, D and Mert, B (2022) Optimization of spray drying for Chlorella vulgaris by using RSM methodology and maltodextrin. Journal of Food Processing and Preservation 46, e16594.Google Scholar
Kruus, M (2017) Purification, Biomass Production and Cryopreservation of Aero-terrestrial Microalgae and Cyanobacteria. Bachelor's thesis, Helsinki Metropolia University of Applied Sciences, p. 47.Google Scholar
Longworth, J, Wu, D, Huete-Ortega, M, Wright, PC and Vaidyanathan, S (2016) Proteome response of Phaeodactylum tricornutum, during lipid accumulation induced by nitrogen depletion. Algal Research 18, 213224.CrossRefGoogle ScholarPubMed
McLellan, MR, Cowling, AJ, Turner, MF and Day, JG (1991) Maintenance of algae and protozoa. In Kirsop, B and Doyle, A (eds), Maintenance of Microorganisms and Cultured Cells. London: Academic Press, pp. 183208.Google Scholar
Mori, F, Erata, M and Watanabe, MM (2002) Cryopreservation of cyanobacteria and green algae. Microbial Culture Collection 18, 4555.Google Scholar
Morris, G (1976) Interactions of rate of cooling, protective additive and warming rate. Archives of Microbiology 107, 5762.CrossRefGoogle ScholarPubMed
Morris, G (1981) Cryopreservation: An Introduction to Cryopreservation in Culture Collections. Cambridge: Institute of Terrestrial Ecology.Google Scholar
Nakanishi, K, Deuchi, K and Kuwano, K (2012) Cryopreservation of four valuable strains of microalgae, including viability and characteristics during 15 years of cryostorage. Journal of Applied Phycology 24, 13811385.CrossRefGoogle Scholar
Odintsova, N and Boroda, A (2012) Cryopreservation of the cells and larvae of marine organisms. Russian Journal of Marine Biology 38, 101111.Google Scholar
Poncet, J-M (2003) Cryopreservation of the unicellular marine alga, Nannochloropsis oculata. Biotechnology Letters 25, 20172022.CrossRefGoogle ScholarPubMed
Racharaks, R and Peccia, J (2019) Cryopreservation of Synechococcus elongatus UTEX 2973. Journal of Applied Phycology 31, 22672276.CrossRefGoogle Scholar
Rhodes, L, Smith, J, Tervit, R, Roberts, R, Adamson, J, Adams, S and Decker, M (2006) Cryopreservation of economically valuable marine micro-algae in the classes Bacillariophyceae, Chlorophyceae, Cyanophyceae, Dinophyceae, Haptophyceae, Prasinophyceae, and Rhodophyceae. Cryobiology 52, 152156.CrossRefGoogle ScholarPubMed
Saadaoui, I, Al Emadi, M, Bounnit, T, Schipper, K and Al Jabri, H (2016) Cryopreservation of microalgae from desert environments of Qatar. Journal of Applied Phycology 28, 22332240.CrossRefGoogle Scholar
Salas-Leiva, JS and Dupré, E (2011) Criopreservación de las microalgas Chaetoceros calcitrans (Paulsen): análisis del efecto de la temperatura de DMSO y régimen de luz durante diferentes períodos de equilibrio. Latin American Journal of Aquatic Research 39, 271279.CrossRefGoogle Scholar
Schmollinger, S, Mühlhaus, T, Boyle, NR, Blaby, IK, Casero, D, Mettler, T, Moseley, JL, Sommer, F, Strenkert, D, Hemme, D, Pellegrini, M, Grossman, AR, Stitt, M, Schroda, M and Merchant, SS (2014) Nitrogen-sparing mechanisms in Chlamydomonas affect the transcriptome, the proteome, and photosynthetic metabolism. The Plant Cell 26, 14101435.CrossRefGoogle ScholarPubMed
Tanniou, A, Turpin, V and Lebeau, T (2012) Comparison of cryopreservation methods for the long term storage of the marine diatom Haslea ostrearia (Simonsen). Cryobiology 65, 4550.CrossRefGoogle Scholar
Taylor, R and Fletcher, RL (1998) Cryopreservation of eukaryotic algae – a review of methodologies. Journal of Applied Phycology 10, 481501.CrossRefGoogle Scholar
Tzovenis, I, Triantaphyllidis, G, Naihong, X, Chatzinikolaou, E, Papadopoulou, K, Xouri, G and Tafas, T (2004) Cryopreservation of marine microalgae and potential toxicity of cryoprotectants to the primary steps of the aquacultural food chain. Aquaculture 230, 457473.CrossRefGoogle Scholar
Xie, D, Ji, X, Zhou, Y, Dai, Y, He, Y, Sun, H, Guo, Z, Yang, Y, Zheng, X and Chen, B (2022) Chlorella vulgaris cultivation in pilot-scale to treat real swine wastewater and mitigate carbon dioxide for sustainable biodiesel production by direct enzymatic transesterification. Bioresource Technology 349, 12688.Google ScholarPubMed