Abstract
The effect of pulsed electric field (PEF) treatments of different intensities on the electroporation of the cytoplasmatic membrane of Chlorella vulgaris, and on the extraction of carotenoids and chlorophylls were investigated. Staining the cells with propidium iodide before and after the PEF treatment revealed the existence of reversible and irreversible electroporation. Application of PEF treatments in the range of 20–25 kV cm−1 caused most of the population of C. vulgaris to be irreversibly electroporated even at short treatment times (5 pulses of 3 µs). However, at lower electric field strengths (10 kV cm−1), cells that were reversibly electroporated were observed even after 50 pulses of 3 µs. The electroporation of C. vulgaris cells by PEF higher than 15 kV cm−1 and duration is higher than 15 µs increased significantly the extraction yield of intracellular components of C. vulgaris. The application of a 20 kV cm−1 for 75 μs increased the extraction yield just after the PEF treatment of the carotenoids, and chlorophylls a and b 0.5, 0.7, and 0.8 times, respectively. However, further increments in electric field strength and treatment time did not cause significant increments in the extraction yield. The extraction of carotenoids from PEF-treated C. vulgaris cells after 1 h of the application of the treatment significantly increased the extraction yield in comparison to the yield obtained from the cells extracted just after the PEF treatment. After PEF treatment at 20 kV cm−1 for 75 µs, extraction yield for carotenoids, and chlorophylls a and b increased 1.2, 1.6, and 2.1 times, respectively. A high correlation was observed between irreversible electroporation and percentage of yield increase when the extraction was conducted after 1 h of the application of PEF treatment (R: 0.93), but not when the extraction was conducted just after PEF treatment (R: 0.67).
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Aronsson K, Ronner U, Borch E (2005) Inactivation of Escherichia coli, Listeria innocua and Saccharomyces cerevisiae in relation to membrane permeabilization and subsequent leakage of intracellular compounds due to pulsed electric field processing. Int J Food Microbiol 99:19–32
Balasundaram B, Harrison S, Bracewell DG (2009) Advances in product release strategies and impact on bioprocess design. Trends Biotechnol 27:477–485
Boussetta N, Grimi N, Lebovka NI, Vorobiev E (2013) “Cold” electroporation in potato tissue induced by pulsed electric field. J Food Eng 115:232–236
Ceron MC, Campos I, Sanchez JF, Acien FG, Molina E, Fernandez-Sevilla JM (2008) Recovery of lutein from microalgae biomass: development of a process for Scenedesmus almeriensis biomass. J Agric Food Chem 56:11761–11766
Cheng C-H, Du T-B, Pi H-C, Jang S-M, Lin Y-H, Lee H-T (2011) Comparative study of lipid extraction from microalgae by organic solvent and supercritical CO2. Bioresour Technol 102:10151–10153
Cooney M, Young G, Nagle N (2009) Extraction of bio-oils from microalgae. Sep Purif Rev 38:291–325
Coustets M, Al-Karablieh N, Thomsen C, Teissie J (2013) Flow process for electroextraction of total proteins from microalgae. J Membr Biol 246:751–760
Del Campo JA, Garcia-Gonzalez M, Guerrero MG (2007) Outdoor cultivation of microalgae for carotenoid production: current state and perspectives. Appl Microbiol Biotechnol 74:1163–1174
Donsi F, Ferrari G, Pataro G (2010) Applications of pulsed electric field treatments for the enhancement of mass transfer from vegetable tissue. Food Eng Rev 2:109–130
Gao Y, Yang M, Wang C (2013) Nutrient deprivation enhances lipid content in marine microalgae. Bioresour Technol 147:484–491
Garcia D, Gomez N, Manas P, Raso J, Pagan R (2007) Pulsed electric fields cause bacterial envelopes permeabilization depending on the treatment intensity, the treatment medium pH and the microorganism investigated. Int J Food Microbiol 113:219–227
Gassel S, Breitenbach J, Sandmann G (2014) Genetic engineering of the complete carotenoid pathway towards enhanced astaxanthin formation in Xanthophyllomyces dendrorhous starting from a high-yield mutant. Appl Microbiol Biotechnol 98:345–350
Goettel M, Eing C, Gusbeth C, Straessner R, Frey W (2013) Pulsed electric field assisted extraction of intracellular valuables from microalgae. Algal Res 2:401–408
Gouveia L, Veloso V, Reis A, Fernandes H, Novais J, Empis J (1996) Evolution of pigment composition in Chlorella vulgaris. Bioresour Technol 57:157
Grimi N, Dubois A, Marchal L, Jubeau S, Lebovka NI, Vorobiev E (2014) Selective extraction from microalgae Nannochloropsis sp. using different methods of cell disruption. Bioresour Technol 153:254–259
Ivorra A (2010) Tissue electroporation as a bioelectric phenomenon: basic concepts. In: Rubinsky B (ed) Irreversible electroporation. Springer, Berlin, pp 23–61
Jeon H, Lee Y, Chang KS, Lee CG, Jin E (2013) Enhanced production of biomass and lipids by supplying CO2 in marine microalga Dunaliella sp. J Microbiol 51:773–776
Lichtenthaler H (1987) Chlorophyll and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382
Macias-Sanchez MD, Fernandez-Sevilla JM, Acien Fernandez FG, Ceron Garcia MC, Molina Grima E (2010) Supercritical fluid extraction of carotenoids from Scenedesmus almeriensis. Food Chem 123:928–935
Mendes RL, Nobre BP, Cardoso MT, Pereira AP, Palavra AF (2003) Supercritical carbon dioxide extraction of compounds with pharmaceutical importance from microalgae. Inorg Chim Acta 356:328–334
Monfort S, Saldana G, Condon S, Raso J, Alvarez I (2012) Inactivation of Salmonella spp. in liquid whole egg using pulsed electric fields, heat, and additives. Food Microbiol 30:393–399
Olaizola M (2003) Commercial development of microalgal biotechnology: from the test tube to the marketplace. Biomol Eng 20:459–466
Prabakaran P, Ravindran AD (2011) A comparative study on effective cell disruption methods for lipid extraction from microalgae. Lett Appl Microbiol 53:150–154
Puertolas E, Luengo E, Alvarez I, Raso J (2012) Improving mass transfer to soften tissues by pulsed electric fields: fundamentals and applications. In: Doyle MP, Klaenhammer TR (eds) Annual review of food science and technology, vol 3. Annual Reviews, Palo Alto, pp 263–282
Pulz O, Gross W (2004) Valuable products from biotechnology of microalgae. Appl Microbiol Biotechnol 6:635–648
Saldana G, Puertolas E, Alvarez I, Meneses N, Knorr D, Raso J (2010) Evaluation of a static treatment chamber to investigate kinetics of microbial inactivation by pulsed electric fields at different temperatures at quasi-isothermal conditions. J Food Eng 100:349–356
Sheng J, Vannela R, Rittmann BE (2011) Evaluation of cell-disruption effects of pulsed-electric-field treatment of Synechocystis PCC 6803. Environ Sci Technol 45:3795–3802
Vanthoor-Koopmans M, Wijffels RH, Barbosa MJ, Eppink MHM (2013) Biorefinery of microalgae for food and fuel. Bioresour Technol 135:142–149
Weaver JC, Chizmadzhev YA (1996) Theory of electroporation: a review. Bioelectrochem Bioenerg 41:135–160
Wouters PC, Bos AP, Ueckert J (2001) Membrane permeabilization in relation to inactivation kinetics of Lactobacillus species due to pulsed electric fields. Appl Environ Microbiol 67:3092–3101
Zbinden MDA, Sturm BSM, Nord RD, Carey WJ, Moore D, Shinogle H, Stagg-Williams SM (2013) Pulsed electric field (PEF) as an intensification pretreatment for greener solvent lipid extraction from microalgae. Biotechnol Bioeng 110:1605–1615
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Luengo, E., Condón-Abanto, S., Álvarez, I. et al. Effect of Pulsed Electric Field Treatments on Permeabilization and Extraction of Pigments from Chlorella vulgaris . J Membrane Biol 247, 1269–1277 (2014). https://doi.org/10.1007/s00232-014-9688-2
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DOI: https://doi.org/10.1007/s00232-014-9688-2