An application of pulsed electric field for intracellular extraction from Chlorella vulgaris TISTR8580
Abstract
This study aimed to apply Pulsed Electric Field (PEF) for Chlorella vulgaris TISTR8580 intracellular extraction. Twenty-five g/L of algal cell samples were treated with PEF at 5 kV/cm under different pulses (1,500 2,500 3,500 and 4,500 pulses) with the initial temperature of 7 degree Celsius. Temperature, pH and conductivity were measured before and after treatments. Total carbohydrate and protein contents in the crude extracts were determined by the Phenol sulfuric acid method and the Bradford assay, respectively. Subsequently, carbohydrate composition was analyzed by Thin Layer Chromatography (TLC). In addition, the morphology of PEF-treated cells was observed under light microscope and cell survival was examined by re-culture in BG-11. The results showed an increased number of PEF pulses raised the temperature and conductivity of the algal extract. The highest total carbohydrate (23.19 ± 1.47 mg/L) and protein (144.48 ± 1.41 mg/L) contents were obtained from the crude extract treated with 2,500 pulses. TLC revealed that disaccharides and oligosaccharides were the major carbohydrate composition in the extract. PEF-treated algal cells exhibited color fading without any cell lysis. However, PEF treatment at 2,500 pulses led to cell death. This study clearly suggested that PEF is a potential technology for C. vulgaris TISTR8580 carbohydrate and protein extraction. Further optimization to enhance PEF extraction efficiency are still needed to achieve high yield of algal intracellular extract. Keywords : microalgae, Pulsed Electric Field, extractionReferences
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Bhat, F. Z., Morton, D. J., Mason, L. S., & Bekhit, A. E. A. (2018). Current and future prospects for the use of pulsed electric field in the meat industry. Critical Reviews in Food Science and Nutrition, 58, 1-15.
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Chittapun, S., Charoenrat, T., Maijui, I., & Antimanon, S. (2017). Development of a simple inclined algal culture system for outdoor cultivation. Science & Technology Asia, 22(3), 1-6.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry Journal, 28, 350–356.
Goettel, M., Eing, C., Gusbeth, C., Straessner, R., & Frey, W. (2013). Pulsed electric field assisted extraction of intracellular valuables from microalgae. Algal Research Journal, 2, 401–408.
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Luengo, E., Martínez, J.M., Bordetas, A., Álvarez, I., & Raso, J. (2015). Influence of the treatment medium temperature on lutein extraction assisted by pulsed electric fields from Chlorella vulgaris. Innovative Food Science and Emerging Technologies, 29, 15–22.
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Postma, P.R., Pataro, G., Capitoli, M., Barbosa, M.J., Wijffels, R.H., Eppink, M.H.M., Olivieri, G., & Ferrari, G. (2016). Selective extraction of intracellular components from the microalga Chlorella vulgaris by combined pulsed electric field–temperature treatment. Bioresource Technology Journal, 203, 80–88.
Panyamuangjai, V., Janthara, S., Kusuya, R., Yawootti, A. & Intra, P. (2012). Juice extraction from fresh apples using pulsed electric field combined with mechanical pressing. Journal of KMUTNB, 4, 469 – 484. (in Thai)
Timmermans, R.A.H., Nierop Groot, M.N., Nederhoff, A.L., van Boekel, M.A.J.S., Matser, A.M., & Mastwijk, H.C., (2014). Pulsed electric field processing of different fruit juices: impact of pH and temperature on inactivation of spoilage and pathogenic micro-organisms. International Journal of Food Microbiology, 173, 105–111.
Vanthoor-Koopmans, M., Wijffels, R.H., Barbosa, M.J., & Eppink, M.H.M., (2013). Biorefinery of microalgae for food and fuel. Bioresource Technology journal, 135, 142–149.
‘t Lam, G.P., Postma, P.R., Fernandes, D.A., Timmermans, R.A.H., Vermuë, M.H., Barbosa, M.J., Eppink, M.H.M., Wijffels, R.H., & Olivieri, G. (2017). Pulsed electric field for protein release of the microalgae Chlorella vulgaris and Neochloris oleoabundans. Algal Research, 24, 181-187
Batista, A.P., Gouveia, L., Bandarra, N.M., Franco, J.M., & Raymundo, A. (2013). Comparison of microalgal biomass profiles as novel functional ingredient for food products. Algal Research Journal, 2, 164–173.
Becker, E.W. (2007). Micro-algae as a source of protein. Biotechnology Advances, 25, 207-210.
Bhat, F. Z., Morton, D. J., Mason, L. S., & Bekhit, A. E. A. (2018). Current and future prospects for the use of pulsed electric field in the meat industry. Critical Reviews in Food Science and Nutrition, 58, 1-15.
Bradford, M.M. (1976). Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.
Chittapun, S., Charoenrat, T., Maijui, I., & Antimanon, S. (2017). Development of a simple inclined algal culture system for outdoor cultivation. Science & Technology Asia, 22(3), 1-6.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry Journal, 28, 350–356.
Goettel, M., Eing, C., Gusbeth, C., Straessner, R., & Frey, W. (2013). Pulsed electric field assisted extraction of intracellular valuables from microalgae. Algal Research Journal, 2, 401–408.
Gouveia, L., Choubert, G., Gomes, E., Rema, P., & Empis, J. (1998). Use of Chlorella vulgaris as a carotenoid source for salmonids: Effect of dietary lipid content on pigmentation, digestibility and muscular retention. Aquaculture International, 6, 269–279.
Grimi, N., Mamouni, F., Lebovka, N., Vorobiev, E., & Vaxelaire, J. (2011). Impact of apple processing modes on extracted juice quality: Pressing assisted by pulsed electric fields. Journal of Food Engineering, 103,
52-61.
Hosikian, A., Lim, S., Halim, R., & Danquah M. K. (2010). Chlorophyll extraction from microalgae: a review on the process engineering aspects. International Journal of Chemical Engineering, 1-11.
Juan, M.M., Luengo, E., Saldaña, G., Álvarez, I., & Raso, J. (2017). C-phycocyanin extraction assisted by pulsed electric field from Artrosphira platensis. Food Research International, 99, 1042 –1047.
Kungsadan, T. (2011). Food Preservation using High Electrical Field Pulse (HELP) Technique. Journal of KMUTNB, 21(1), 198 – 207. (in Thai)
Laemmli, U.K. (1970). Cleavage of structure protein during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
Lichtenthaler, H.K., & Buschmann, C. (2001). Chlorophylls and carotenoids: measurement and characterization by UV-VIS spectroscopy. Current Protocols in Food Analytical Chemistry, F4.3.1-F4.3.8.
Lindgren, M., Aronsson, K., Galt, S., & Ohlsson, T. (2002). Simulation of the temperature increase in pulsed electric field (PEF) continuous flow treatment chambers. Innovative Food Science and Emerging Technologies, 3, 233–245.
Luengo, E., Martínez, J.M., Bordetas, A., Álvarez, I., & Raso, J. (2015). Influence of the treatment medium temperature on lutein extraction assisted by pulsed electric fields from Chlorella vulgaris. Innovative Food Science and Emerging Technologies, 29, 15–22.
Oleksii, P., Francisco, J. B., Nabil, G., Luc, M., Sébastien, J., Nikolai, L., & Eugene, V. (2015). Pulsed electric field and pH assisted selective extraction of intracellular components from microalgae Nannochloropsis. Algal Research, 8,128-134.
Postma, P.R., Miron, T.L., Olivieri, G., Barbosa, M.J., Wijffels, R.H., & Eppink, M.H.M. (2015). Mild disintegration of the green microalgae Chlorella vulgaris using bead milling. Bioresource Technology Journal, 184,
297–304.
Postma, P.R., Pataro, G., Capitoli, M., Barbosa, M.J., Wijffels, R.H., Eppink, M.H.M., Olivieri, G., & Ferrari, G. (2016). Selective extraction of intracellular components from the microalga Chlorella vulgaris by combined pulsed electric field–temperature treatment. Bioresource Technology Journal, 203, 80–88.
Panyamuangjai, V., Janthara, S., Kusuya, R., Yawootti, A. & Intra, P. (2012). Juice extraction from fresh apples using pulsed electric field combined with mechanical pressing. Journal of KMUTNB, 4, 469 – 484. (in Thai)
Timmermans, R.A.H., Nierop Groot, M.N., Nederhoff, A.L., van Boekel, M.A.J.S., Matser, A.M., & Mastwijk, H.C., (2014). Pulsed electric field processing of different fruit juices: impact of pH and temperature on inactivation of spoilage and pathogenic micro-organisms. International Journal of Food Microbiology, 173, 105–111.
Vanthoor-Koopmans, M., Wijffels, R.H., Barbosa, M.J., & Eppink, M.H.M., (2013). Biorefinery of microalgae for food and fuel. Bioresource Technology journal, 135, 142–149.
‘t Lam, G.P., Postma, P.R., Fernandes, D.A., Timmermans, R.A.H., Vermuë, M.H., Barbosa, M.J., Eppink, M.H.M., Wijffels, R.H., & Olivieri, G. (2017). Pulsed electric field for protein release of the microalgae Chlorella vulgaris and Neochloris oleoabundans. Algal Research, 24, 181-187
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2018-09-12
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