Аннотация

Microalgae are a source of biologically active substances, e.g., polysaccharides. Their commercial potential attracts a lot of scientific attention. This research featured the effect of various nutrient media on the biomass of the psychrophilic microalga Skeletonema pseudocostatum and its ability to synthesize polysaccharides.
The Tamiya nutrient medium was used as the standard one. Its composition was improved to accelerate biomass cultivation. Optimization principles were based on the unconventional mathematical method for multidimensional modeling and involved the ANETR 21 software. The qualitative and quantitative assessment of microalgal polysaccharides relied on the anthrone sulfate method. The concentration of uronic acids was determined by the carbazole method while neutral sugars were studied by the resorcinol sulfate method in a microalgal suspension. The growth index of the S. pseudocostatum biomass was represented as a ratio of the maximal mass to the initial mass.
The maximal growth index was achieved by adding to the standard Tamiya medium: 10.00 g/cm³ potassium nitrate (MgSO₄×7H₂O), 2.50 g/cm³ potassium dihydrogen phosphate, 1 cm³ Fe⁺ ethylenediaminetetraacetic acid (EDTA), and a mix of 4.29 g/cm³ boric acid and 0.9 g/cm³ manganese (II) chloride tetrahydrate (MnCl₂×4H₂O) in an amount of 1.00 cm³. The maximal polysaccharide biosynthesis was observed when the nutrient medium was modified as follows: 5.00 g/m³ potassium nitrate, 3.75 g/cm³ magnesium sulfate, 2.50 g/cm³ potassium dihydrogen phosphate, 1 mm³ solution of Fe⁺ ethylenediaminetetraacetic acid, and solution of 1.0 g/cm3 boric acid and 1.81 g/cm³ MnCl₂×4H₂O (1.00 mm³ each). The maximal accumulation of microalgal biomass was 2.88 ± 0.08 μg/100 mg dry solids; the maximal yield of polysaccharides was 3.16 ± 0.09 μg/100 mg dry solids. These results were obtained at 5°C.
The yield of polysaccharides by S. pseudocostatum depended on such cultivation parameters as temperature and pH. At cultivation temperatures of 0, 5, and 10°C, the yield of polysaccharides reached 2.13 ± 0.06, 3.16 ± 0.09, and 2.04 ± 0.06 μg/100 mg dry solids, respectively. The yield of exopolysaccharides represented by uronic acids and neutral sugars was 106.3 ± 3.1 mg/g and 806.6 ± 24.0 mg/g, respectively. In this research, polysaccharides synthesized by S. pseudocostatum demonstrated good prospects for the food industry and sustainable organic agriculture.

Ключевые слова

Microalgae, Skeletonema pseudocostatum, polysaccharides, optimization parameters, nutrient medium

ФИНАНСИРОВАНИЕ

The research was supported by the Ministry of Science and Higher Education of the Russian Federation as part of grant from the President of the Russian Federation, project no. MK-484.2022.1.4 (agreement no. 075-15-2022-393).

СПИСОК ЛИТЕРАТУРЫ

1. de Carvalho Silvello MA, Gonçalves IS, Azambuja SPH, Costa SS, Silva PGP, et al. Microalgae-based carbohydrates: A green innovative source of bioenergy. Bioresource Technology. 2022;344(Part B):126304. https://doi.org/10.1016/j.biortech.2021.126304
2. Dolganyuk V, Belova D, Babich O, Prosekov A, Ivanova S, et al. Microalgae: A promising source of valuable bioproducts. Biomolecules. 2020;10(8):1153. https://doi.org/10.3390/biom10081153
3. Hu J, Nagarajan D, Zhang Q, Chang JS, Lee DJ. Heterotrophic cultivation of microalgae for pigment production: A review. Biotechnology Advances. 2018;36(1):54–67. https://doi.org/10.1016/j.biotechadv.2017.09.009
4. Dolganyuk V, Sukhikh S, Kalashnikova O, Ivanova S, Kashirskikh E, et al. Food Proteins: Potential resources. Sustainability. 2023;15(7):5863. https://doi.org/10.3390/su15075863
5. Gonçalves AL. The use of microalgae and cyanobacteria in the improvement of agricultural practices: A review on their Biofertilising, Biostimulating and biopesticide roles. Applied Sciences. 2021;11(2):871. https://doi.org/10.3390/app11020871
6. López-Hernández JF, Kean-Meng T, Asencio-Alcudia GG, Asyraf-Kassim M, Alvarez-González CA, et al. Sustainable microalgae and cyanobacteria biotechnology. Applied Sciences. 2022;12(14):6887. https://doi.org/10.3390/app12146887
7. Coulombier N, Blanchier P, Le Dean L, Barthelemy V, Lebouvier N, et al. The effects of CO2-induced acidification on Tetraselmis biomass production, photophysiology and antioxidant activity: A comparison using batch and continuous culture. Journal of Biotechnology. 2021;325:312–324. https://doi.org/10.1016/j.jbiotec.2020.10.005
8. Paliwal C, Mitra M, Bhayani K, Bharadwaj SVV, Ghosh T, et al. Abiotic stresses as tools for metabolites in microalgae. Bioresource Technology. 2017;244(Part 2):1216–1226. https://doi.org/10.1016/j.biortech.2017.05.058
9. Marques AEML, Balen RE, da Silva Pereira Fernandes L, Motta CM, de Assis HCS, et al. Diets containing residual microalgae biomass protect fishes against oxidative stress and DNA damage. Journal of Applied Phycology. 2019;31:2933–2940. https://doi.org/10.1007/s10811-019-01825-6
10. Gürlek C, Yarkent Ç, Köse A, Tuğcu B, Gebeloğlu IK, et al. Screening of antioxidant and cytotoxic activities of several microalgal extracts with pharmaceutical potential. Health and Technology. 2020;10:111–117. https://doi.org/10.1007/s12553-019-00388-3
11. Chaplygina TV, Prosekov AU, Babich OO, Pavsky VA, Ivanova SA. Functional dairy products are protection during pandemic. Dairy industry. 2020;(6):26–28. https://doi.org/10.31515/1019-8946-2020-06-26-28
12. Morais MG, Santos TD, Moraes L, Vaz BS, Morais EG, et al. Exopolysaccharides from microalgae: Production in a biorefinery framework and potential applications. Bioresource Technology Reports. 2022;18:101006. https://doi.org/10.1016/j.biteb.2022.101006.
13. Chanda MJ, Merghoub N, Arroussi HE. Microalgae polysaccharides: The new sustainable bioactive products for the development of plant bio-stimulants? World Journal of Microbiology and Biotechnology. 2019;35:177. https://doi.org/10.1007/s11274-019-2745-3
14. Moreira JB, Kuntzler SG, Bezerra PQM, Cassuriaga APA, Zaparoli M, et al. Recent advances of microalgae exopolysaccharides for application as bioflocculants. Polysaccharides 2022;3(1):264–276. https://doi.org/10.3390/polysaccharides3010015
15. Gerecht A, Romano G, Ianora A, d’Ippolito G, Cutignano A, et al. Plasticity of oxylipin metabolism among clones of the marine diatom Skeletonema marinoi (Bacillariophyceae). Journal of Phycology. 2011;47(5):1050–1056. https://doi.org/10.1111/j.1529-8817.2011.01030.x
16. McKenzie CH, Bates SS, Martin JL, Haigh N, Howland KL, et al. Three decades of Canadian marine harmful algal events: Phytoplankton and phycotoxins of concern to human and ecosystem health. Harmful Algae. 2021;102:101852. https://doi.org/10.1016/j.hal.2020.101852
17. Kooistra WHCF, Sarno D, Balzano S, Gu H, Andersen RA, et al. Global diversity and biogeography of Skeletonema species (Bacillariophyta). Protist. 2008;159(2):177–193. https://doi.org/10.1016/j.protis.2007.09.004
18. Aremu AO, Neményi M, Stirk WA, Ördög V, van Staden J. Manipulation of nitrogen levels and mode of cultivation are viable methods to improve the lipid, fatty acids, phytochemical content, and bioactivities in Chlorella minutissima. Journal of Phycology. 2015;51(4):659–669. https://doi.org/10.1111/jpy.12308
19. Coulombier N, Jauffrais T, Lebouvier N. Antioxidant compounds from microalgae: A Review. Marine Drugs.2021;19(10):549. https://doi.org/10.3390/md19100549
20. Paradis-Hautcoeur J, Gosselin M, Villeneuve V, Tremblay J, Lévesque D, et al. Effects of riverine nutrient inputs on the sinking fluxes of microbial particles in the St. Lawrence Estuary. Estuarine, Coastal and Shelf Science. 2023;284:108270. https://doi.org/10.1016/j.ecss.2023.108270
21. Azizan A, Bustamam MSA, Maulidiani M, Shaari K, Ismail IS, et al. Metabolite profiling of the microalgal diatom chaetoceros calcitrans and correlation with antioxidant and nitric oxide inhibitory activities via 1H NMR-based metabolomics. Marine Drugs. 2018;16(5):154. https://doi.org/10.3390/md16050154
22. Jaroszuk-Ściseł J, Nowak A, Komaniecka I, Choma A, et al. Differences in production, composition, and antioxidant activities of exopolymeric substances (EPS) obtained from cultures of endophytic Fusarium culmorum strains with different effects on cereals. Molecules. 2020;25(3):616. https://doi.org/10.3390/molecules25030616
23. Koçer AT, İnan B, Usul SK, Özçimen D, Yılmaz MT, et al. Exopolysaccharides from microalgae: Production, characterization, optimization and techno-economic assessment. Brazilian Journal of Microbiology. 2021;52:1779–1790. https://doi.org/10.1007/s42770-021-00575-3
24. Ahmed Z, Wang Y, Anjum N, Ahmad H, Ahmad A, et al. Characterization of new exopolysaccharides produced by coculturing of L. kefiranofaciens with yoghurt strains. International Journal of Biological Macromolecules. 2013;59:377–383. https://doi.org/10.1016/j.ijbiomac.2013.04.075
25. Martirosyan VV, Kostyuchenko MN, Reynov MV, Tyurina OE, Savkina OA. Study of exopolysaccharide production by lactic acid bacteria used in the baking industry and comparison of methods for their determination. Food Metaengineering. 2024;2(4):13–25. (In Russ.) https://doi.org/10.37442/fme.2024.4.67
26. Pérez-Lechuga G, Venegas-Martínez F, Martínez-Sánchez JF. Mathematical modeling of manufacturing lines with distribution by process: A Markov chain approach. Mathematics. 2021;9(24):3269. https://doi.org/10.3390/math9243269
27. Papuga S, Pećanac I, Stojković M, Savić A, Velemir A. Mead fermentation parameters: Optimization by response surface methodology. Foods and Raw Materials. 2022;10(1):137–147. https://doi.org/10.21603/23084057-2022-1-137-147
28. Taylor KA, Buchanan-Smith JG. A colorimetric method for the quantitation of uronic acids and a specific assay for galacturonic acid. Analytical Biochemistry. 1992;201(1):190–196. https://doi.org/10.1016/0003-2697(92)90194-C
29. Monsigny M, Petit C, Roche AC. Colorimetric determination of neutral sugars by a resorcinol sulfuric acid micromethod. Analytical Biochemistry. 1988;175(2):525–530. https://doi.org/10.1016/0003-2697(88)90578-7
30. Myklestad S. Production of carbohydrates by marine planktonic diatoms. II. Influence of the NP ratio in the growth medium on the assimilation ratio, growth rate, and production of cellular and extracellular carbohydrates by Chaetoceros affinis var. willei (Gran) Hustedt and Skeletonema costatum (Grev.) Cleve. Journal of Experimental Marine Biology and Ecology. 1977;29(2):161–179. https://doi.org/10.1016/0022-0981(77)90046-6
31. Taraldsvik M, Myklestad S. The effect of pH on growth rate, biochemical composition and extracellular carbohydrate production of the marine diatom Skeletonema costatum. European Journal of Phycology 2000;35(2):189–194. https://doi.org/10.1080/09670260010001735781
32. Granum E, Kirkvold S, Myklestad SM. Cellular and extracellular production of carbohydrates and amino acids by the marine diatom Skeletonema costatum: Diel variations and effects of N depletion. Marine Ecology-progress Series. 2002;242:83–94. https://doi.org/10.3354/meps242083
33. Ocaranza D, Balic I, Bruna T, Moreno I, et al. A modeled high-density fed-batch culture improves biomass growth and β-glucans accumulation in Microchloropsis salina. Plants. 2022;11(23):3229. https://doi.org/10.3390/plants11233229
34. Sathasivam R, Ki J. A review of the biological activities of microalgal carotenoids and their potential use in healthcare and cosmetic industries. Marine Drugs. 2018;16(1):26. https://doi.org/10.3390/md16010026
35. Yue F, Zhang J, Xu J, Niu T, Lü X, et al. Effects of monosaccharide composition on quantitative analysis of total sugar content by phenol-sulfuric acid method. Frontiers in Nutrition. 2022;9:963318. http://doi.org/10.3389/fnut.2022.963318