Microwave-Assisted Extraction of Polysaccharides from Chlorella pyrenoidosa and Its Characterization

https://doi.org/10.22146/ijc.103337

Margaretha Praba Aulia(1), Muhammad Mufti Azis(2), Rochmadi Rochmadi(3), Arief Budiman(4*)

(1) Department Chemical Engineering, Universitas Gadjah Mada, Jl. Grafika No. 2, Yogyakarta 55284, Indonesia; Department Agrotechnology, Faculty of Agriculture and Animal Husbandry, Universitas Boyolali, Jl. Pandanaran No. 405, Winong, Boyolali 57315, Indonesia
(2) Department Chemical Engineering, Universitas Gadjah Mada, Jl. Grafika No. 2, Yogyakarta 55284, Indonesia
(3) Department Chemical Engineering, Universitas Gadjah Mada, Jl. Grafika No. 2, Yogyakarta 55284, Indonesia
(4) Department Chemical Engineering, Universitas Gadjah Mada, Jl. Grafika No. 2, Yogyakarta 55284, Indonesia; Center of Excellence for Microalgae Biorefinery, Center for Energy Studies (PSE), Universitas Gadjah Mada, Sekip No. K1A, Yogyakarta 55281, Indonesia
(*) Corresponding Author

Abstract


This study explored an efficient method for extracting polysaccharides from Chlorella pyrenoidosa using microwave-assisted extraction (MAE) with water as the solvent, a technique consistent with green chemistry principles. The goal was to enhance the yield and quality of polysaccharides for their potential applications as multifunctional active ingredients in the pharmaceutical and functional food industries. Key extraction parameters, including extraction time (10, 20, and 30 min), temperature (80 °C), and solid-to-liquid ratios (1:20, 1:30, 1:40 m/v), were systematically evaluated. The results indicated that a solid-to-liquid ratio of 1:40 m/v at 80 °C for 10 min yielded the highest polysaccharide content (56.64%). FTIR analysis confirmed the presence of pyranose rings in D-glucose and hydroxyl groups, while HPLC identified D-mannose (58.12%) as the predominant sugar, followed by D-glucose (34.46%), D-galactose (3.61%), and L-rhamnose (3.81%). Purified polysaccharide was composed of major mannose and glucose, a biomolecule very important given that it has wide applications in medical and food industries.


Keywords


Chlorella pyrenoidosa; glucose; mannose; microwave-assisted extraction; polysaccharide

Full Text:

Full Text PDF


References

[1] Bussa, M., Eisen, A., Zollfrank, C., and Röder, H., 2019, Life cycle assessment of microalgae products: State of the art and their potential for the production of polylactid acid, J. Cleaner Prod., 213, 1299–1312.

[2] Hildebrand, G., Poojary, M.M., O’Donnell, C., Lund, M.N., Garcia-Vaquero, M., and Tiwari, B.K., 2020, Ultrasound-assisted processing of Chlorella vulgaris for enhanced protein extraction, J. Appl. Phycol., 32 (3), 1709–1718.

[3] Guo, W., Zhu, S., Li, S., Feng, Y., Wu, H., and Zeng, M., 2021, Microalgae polysaccharides ameliorates obesity in association with modulation of lipid metabolism and gut microbiota in high-fat-diet fed C57BL/6 mice, Int. J. Biol. Macromol., 182, 1371–1383.

[4] Khobragade, T.P., Giri, P., Pagar, A.D., Patil, M.D., Sarak, S., Joo, S., Goh, Y., Jung, S., Yoon, H., Yun, S., Kwon, Y., and Yun, H., 2023, Dual-function transaminases with hybrid nanoflower for the production of value-added chemicals from biobased levulinic acid, Front. Bioeng. Biotechnol., 11, 1280464.

[5] Signoretto, M., Taghavi, S., Ghedini, E., and Menegazzo, F., 2019, Catalytic production of levulinic acid (LA) from actual biomass, Molecules, 24 (15), 2760.

[6] Prasad, R.K., Chatterjee, S., Mazumder, P.B., Gupta, S.K., Sharma, S., Vairale, M.G., Datta, S., Dwivedi, S.K., and Gupta, D.K., 2019, Bioethanol production from waste lignocelluloses: A review on microbial degradation potential, Chemosphere, 231, 588–606.

[7] Jamilatun, S., Budhijanto, B., Rochmadi, R., Yuliestyan, A., Hadiyanto, H., and Budiman, A., 2019, Comparative analysis between pyrolysis products of Spirulina platensis biomass and its residues, Int. J. Renewable Energy Dev., 8 (2), 133–140.

[8] Jamilatun, S., Budiman, A., Budhijanto, B., and Rochmadi, R., 2017, Non-catalytic slow pyrolysis of Spirulina platensis residue for production of liquid biofuel, Int. J. Renewable Energy Dev., 7 (4), 1901–1908.

[9] Liu, F., Chen, H., Qin, L., Al-Haimi, A.A.N.M., Xu, J., Zhou, W., Zhu, S., and Wang, Z., 2023, Effect and characterization of polysaccharides extracted from Chlorella sp. by hot-water and alkali extraction methods, Algal Res., 70, 102970.

[10] Shi, L., 2016, Bioactivities, isolation and purification methods of polysaccharides from natural products: A review, Int. J. Biol. Macromol., 92, 37–48.

[11] Chen, Y., Liu, X., Wu, L., Tong, A., Zhao, L., Liu, B., and Zhao, C., 2018, Physicochemical characterization of polysaccharides from Chlorella pyrenoidosa and its anti-ageing effects in Drosophila melanogaster, Carbohydr. Polym., 185, 120–126.

[12] Yuan, Q., Li, H., Wei, Z., Lv, K., Gao, C., Liu, Y., and Zhao, L., 2020, Isolation, structures and biological activities of polysaccharides from Chlorella: A review, Int. J. Biol. Macromol., 163, 2199–2209.

[13] Lv, K., Yuan, Q., Li, H., Li, T., Ma, H., Gao, C., Zhang, S., Liu, Y., and Zhao, L., 2022, Chlorella pyrenoidosa polysaccharides as a prebiotic to modulate gut microbiota: Physicochemical properties and fermentation characteristics in vitro, Foods, 11 (5), 725.

[14] Morillas-España, A., Lafarga, T., Sánchez-Zurano, A., Acién-Fernández, F.G., and González-López, C., 2022, Microalgae based wastewater treatment coupled to the production of high value agricultural products: Current needs and challenges, Chemosphere, 291, 132968.

[15] Moreira, J.B., Vaz, B.S., Cardias, B.B., Cruz, C.G., de Almeida, A.C.A., Costa, J.A.V., and de Morais, M.G., 2022, Microalgae polysaccharides: An alternative source for food production and sustainable agriculture, Polysaccharides, 3 (2), 441–457.

[16] Gitau, M.M., Farkas, A., Ördög, V., and Maróti, G., 2022, Evaluation of the biostimulant effects of two Chlorophyta microalgae on tomato (Solanum lycopersicum), J. Cleaner Prod., 364, 132689.

[17] Chick, H., 1903, A study of a unicellular green alga, occurring in polluted water, with especial reference to its nitrogenous metabolism, Proc. R. Soc. London, 71, 458–476.

[18] Thirugnanasambandham, K., Sivakumar, V., and Maran, J.P., 2015, Microwave-assisted extraction of polysaccharides from mulberry leaves, Int. J. Biol. Macromol., 72, 1–5.

[19] Kaderides, K., Papaoikonomou, L., Serafim, M., and Goula, A.M., 2019, Microwave-assisted extraction of phenolics from pomegranate peels: Optimization, kinetics, and comparison with ultrasounds extraction, Chem. Eng. Process., 137, 1–11.

[20] Yahaya, N., Mohamed, A.H., Sajid, M., Zain, N.N.M., Liao, P.C., and Chew, K.W., 2024, Deep eutectic solvents as sustainable extraction media for extraction of polysaccharides from natural sources: Status, challenges and prospects, Carbohydr. Polym., 338, 122199.

[21] Saman, W.R., Mulyadi, A.F., and Wijana, S., 2023, Microwave assisted extraction of beracyanin from peel red dragon fruit (Hylocereus polyrhizus) (study of the extraction time and the ratio of materials: solvent), J. Agercolere, 5 (1), 17–25.

[22] Rahman, S.S.A., Pasupathi, S., Venkatachalam, P., Jothi, A., and Karuppiah, S., 2025, Modeling, optimization, and characterization of polysaccharides from Strychnos potatorum using microwave-assisted extraction, Biomass Convers. Biorefin., 15 (2), 2111–2129.

[23] Ajami, M.R., Ganjloo, A., and Bimakr, M., 2023, Continuous fast microwave-assisted extraction of radish leaves polysaccharides: Optimization, preliminary characterization, biological, and techno-functional properties, Biomass Convers. Biorefin., 13 (16), 14987–15000.

[24] Yuan, Y., and Macquarrie, D., 2015, Microwave assisted extraction of sulfated polysaccharides (fucoidan) from Ascophyllum nodosum and its antioxidant activity, Carbohydr. Polym., 129, 101–107.

[25] Suárez, E.R., Syvitski, R., Kralovec, J.A., Noseda, M.D., Barrow, C.J., Ewart, H.S., Lumsden, M.D., and Grindley, T.B., 2006, Immunostimulatory polysaccharides from chlorella pyrenoidosa. A new galactofuranan. Measurement of molecular weight and molecular weight dispersion by DOSY NMR. Biomacromolecules, 7 (8), 2368–2376.

[26] Hu, Z., Ma, X., and Chen, C., 2012, A study on experimental characteristic of microwave-assisted pyrolysis of microalgae, Bioresour. Technol., 107, 487–493.

[27] Sheng, J., Yu, F., Xin, Z., Zhao, L., Zhu, X., and Hu, Q., 2007, Preparation, identification and their antitumor activities in vitro of polysaccharides from Chlorella pyrenoidosa, Food Chem., 105 (2), 533–539.

[28] Ai, X., Yu, P., Li, X., Lai, X., Yang, M., Liu, F., Luan, F., and Meng, X., 2023, Polysaccharides from Spirulina platensis: Extraction methods, structural features and bioactivities diversity, Int. J. Biol. Macromol., 231, 123211.

[29] Nielsen, S.S., 2024, “Total Carbohydrate by Phenol-Sulfuric Acid Method” in Nielsen's Food Analysis Laboratory Manual, Eds. Ismail, B.P., and Nielsen, S.S., Springer International Publishing, Cham, Switzerland, 147–151.

[30] Li, T.T., Huang, Z.R., Jia, R.B., Lv, X.C., Zhao, C., and Liu, B., 2021, Spirulina platensis polysaccharides attenuate lipid and carbohydrate metabolism disorder in high-sucrose and high-fat diet-fed rats in association with intestinal microbiota, Food Res. Int., 147, 110530.

[31] Tang, Z., Xu, Y., Cai, C., and Tan, Z., 2023, Extraction of Lycium barbarum polysaccharides using temperature-switchable deep eutectic solvents: A sustainable methodology for recycling and reuse of the extractant, J. Mol. Liq., 383, 122063.

[32] Nielsen, S.S., 2010, "Phenol-Sulfuric Acid Method for Total Carbohydrates" in Food Analysis Laboratory Manual, Springer US, Boston, MA, US, 47–53.

[33] Sridhar, A., Ponnuchamy, M., Kumar, P.S., Kapoor, A., Vo, D.V.N., and Prabhakar, S., 2021, Techniques and modeling of polyphenol extraction from food: A review, Environ. Chem. Lett., 19 (4), 3409–3443.

[34] Jamilatun, S., Budhijanto, B., Rochmadi, R., and Budiman, A., 2017, Thermal decomposition and kinetic studies of pyrolysis of Spirulina platensis residue, Int. J. Renewable Energy Dev., 6 (3), 193–201.

[35] Ringgani, R., Azis, M.M., Rochmadi, R., and Budiman, A., 2022, Kinetic study of levulinic acid from Spirulina platensis residue, Appl. Biochem. Biotechnol., 194 (6), 2684–2699.

[36] Ma, Y., Wang, J., Fei, P., Wan, P., Li, C., Yang, L., and Shi, R., 2024, Extraction, characterization and antioxidant activity evaluation of polysaccharides from Chlorella sp, J. Food Meas. Charact., 18 (12), 10080–10092.

[37] Jamilatun, S., Elisthatiana, Y., Aini, S.N., Mufandi, I., and Budiman, A., 2020, Effect of temperature on yield product and characteristics of bio-oil from pyrolysis of Spirulina platensis residue, Elkawnie, 6 (1), 96–108.

[38] Colusse, G.A., Carneiro, J., Duarte, M.E.R., de Carvalho, J.C., and Noseda, M.D., 2022, Advances in microalgal cell wall polysaccharides: A review focused on structure, production, and biological application, Crit. Rev. Biotechnol., 42 (4), 562–577.

[39] Mane, S., Singh, A., and Taneja, N.K., 2025, Pretreatment optimization for microalgae oil yield enhancement and residual biomass characterization for sustainable biofuel feedstock production, Biomass Convers. Biorefinery, 15 (7), 9859–9874.

[40] da Silveira, P.H.P.M., dos Santos, M.C.C., Chaves, Y.S., Ribeiro, M.P., Marchi, B.Z., Monteiro, S.N., Gomes, A.V., Tapanes, N.L.C.O., Pereira, P.S.C., and Bastos, D.C., 2023, Characterization of thermo-mechanical and chemical properties of polypropylene/hemp fiber biocomposites: Impact of maleic anhydride compatibilizer and fiber content, Polymers, 15 (15), 3271.

[41] Adhiputra, R., Utami, M., Suyono, E.A., Budiman, A., Hariani, P.L., Pratiwi, A.S., and Wijaya, K., 2021, Simultaneous extraction and in-situ transesterification of Chlorella vulgaris using microwave-assisted method for biodiesel production, Korean J. Mater. Res., 31 (4), 181–187.

[42] Yu, M., Chen, M., Gui, J., Huang, S., Liu, Y., Shentu, H., He, J., Fang, Z., Wang, W., and Zhang, Y., 2019, Preparation of Chlorella vulgaris polysaccharides and their antioxidant activity in vitro and in vivo, Int. J. Biol. Macromol., 137, 139–150.

[43] Becker, E.W., 2007, Micro-algae as a source of protein, Biotechnol. Adv., 25 (2), 207–210.

[44] Henry, E.C., 2004, Handbook of microalgal culture: biotechnology and applied phycology, J. Phycol., 40 (5), 1001–1002.

[45] Chanda, M.J., Merghoub, N., and EL Arroussi, H., 2019, Microalgae polysaccharides: The new sustainable bioactive products for the development of plant bio-stimulants?, World J. Microbiol. Biotechnol., 35 (11), 177.

[46] Peng, H., Xv, X., Cui, X., Fu, Y., Zhang, S., Wang, G., Chen, X., and Song, W., 2023, Physicochemical characterization and antioxidant activity of polysaccharides from Chlorella sp. by microwave-assisted enzymatic extraction, Front. Bioeng. Biotechnol., 11, 1264641.

[47] Patel, A.K., Vadrale, A.P., Singhania, R.R., Michaud, P., Pandey, A., Chen, S.J., Chen, C.W., and Dong, C.D., 2023, Algal polysaccharides: Current status and future prospects, Phytochem. Rev., 22 (4), 1167–1196.

[48] Mathlouthi, M., and Koenig, J.L., 1987, Vibrational spectra of carbohydrates, Adv. Carbohydr. Chem. Biochem., 44, 7–89.

[49] Kačuráková, M., and Wilson, R.H., 2001, Developments in mid-infrared FT-IR spectroscopy of selected carbohydrates, Carbohydr. Polym., 44 (4), 291–303.

[50] Chen, L., Song, D., Tian, Y., Ding, L., Yu, A., and Zhang, H., 2008, Application of on-line microwave sample-preparation techniques, TrAC, Trends Anal. Chem., 27 (2), 151–159.

[51] Chaiklahan, R., Chirasuwan, N., Loha, V., Tia, S., and Bunnag, B., 2018, Stepwise extraction of high-value chemicals from Arthrospira (Spirulina) and an economic feasibility study, Biotechnol. Rep., 20, e00280.

[52] Ratna, R., Arahman, N., Munawar, A.A., and Aprilia, S., 2023, Extraction, isolation, and characterization of nanocrystalline cellulose from barangan banana (Musa acuminata L.) peduncles waste, Indones. J. Chem., 23 (1), 73–89.



DOI: https://doi.org/10.22146/ijc.103337

Article Metrics

Abstract views : 7897 | views : 3064


Copyright (c) 2025 Indonesian Journal of Chemistry

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

 


Indonesian Journal of Chemistry (ISSN 1411-9420 /e-ISSN 2460-1578) - Chemistry Department, Universitas Gadjah Mada, Indonesia.

Web
Analytics View The Statistics of Indones. J. Chem.