Uncovering the Potential of Nitrogen and Salt Stress for Enhanced ꞵ-Carotene Production and Antioxidant Capacity in Plant Pathogenic Alga Cephaleuros
Article Sidebar
Main Article Content
Abstract
A pure strain of Cephaleuros alga, designated Cephaleuros Cp.1, was successfully isolated directly from a citrus leaf lesion. This study investigated factors influencing both biomass and carotenoid accumulation in this green filamentous alga. Different nitrogen sources, NaCl stress, and trace elements in HSM, BBM, and Bristol media were compared. The autotrophic condition with HSM medium clearly offered the highest green biomass. Interestingly, Cephaleuros Cp.1 remained green in HSM using NH4Cl as the nitrogen source but visibly changed to an orange hue due to the accumulation of β-carotene in BBM containing NaNO3. This color change, along with the lower biomass and more intense yellow color when using nitrate, was the first reported in Cephaleuros, implying that nitrate may cause stress in the alga. Similar phenomena were clearly observed when NaCl was applied to HSM and BBM; on the other hand, Hutner’s trace elements and trace metal solution had no significant effect. These findings suggest, for the first time, a link between stress conditions and the accumulation of β-carotene in Cephaleuros Cp.1. TLC revealed β-carotene as the main carotenoid accumulated by this alga. The accumulation was further enhanced by both nitrogen deficiency and salt stress. However, these stresses also led to a decrease in algal biomass. This study is the first to report free radical scavenging activity linked to β-carotene in Cephaleuros. Among the tested cultures, BBM exhibited the strongest activity (EC50 1.40 mg/mL). These findings hold promise for future applications of Cephaleuros as a source of natural β-carotene with antioxidant properties.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Huang JJ, Lin S, Xu W, Cheung PCK. Occurrence and biosynthesis of carotenoids in phytoplankton. Biotechnol Adv. 2017 Sep 1;35(5):597–618.
Takaichi S. Carotenoids in algae: distributions, biosyntheses and functions. Mar Drugs. 2011 Jun;9(6):1101–18.
Lopez-Bautista JM. The Trentepohliales revisited. Louisiana State University; 2002.
Thompson RH, Wujek DE. Trentepohliales: Cephaleuros, Phycopeltis, and Stomatochroon. Morphology, taxonomy, and ecology. Enfield, New Hampshire (US): Science Publishers; 1997.
Barone ME, Parkes R, Herbert H, McDonnell A, Conlon T, Aranyos A, et al. Comparative response of marine microalgae to H2O2-induced oxidative Stress. Appl Biochem Biotechnol. 2021;193(12):4052–67.
Parkes R, Barone ME, Herbert H, Gillespie E, Touzet N. Antioxidant activity and carotenoid content responses of three Haematococcus sp. (Chlorophyta) strains Exposed to Multiple Stressors. Appl Biochem Biotechnol. 2022;194(10):4492–510.
Shi TQ, Wang LR, Zhang ZX, Sun XM, Huang H. Stresses as first-line tools for enhancing lipid and carotenoid production in microalgae. Front Bioeng Biotechnol. 2020;8:610.
Enwereuzoh UO, Onyeagoro GN. A novel aeration method for the preparation of algae (Dunaliella Salina) biomass for biofuel production. Am J Eng Res. 2014;3(9):209–14.
Christian D, Zhang J, Sawdon AJ, Peng CA. Enhanced astaxanthin accumulation in Haematococcus pluvialis using high carbon dioxide concentration and light illumination. Bioresour Technol. 2018;256:548–51.
Wolf L, Cummings T, Müller K, Reppke M, Volkmar M, Weuster-Botz D. Production of β-carotene with Dunaliella salina CCAP19/18 at physically simulated outdoor conditions. Eng Life Sci. 2021;21(3–4):115–25.
Suto Y, Ohtani S. Morphology and taxonomy of five Cephaleuros species (Trentepohliaceae, Chlorophyta) from Japan, including three new species. Phycologia. 2009;48(4):213–36.
Sueoka N, Chiang KS, Kates JR. Deoxyribonucleic acid replication in meiosis of Chlamydomonas reinhardi: I. isotopic transfer experiments with a strain producing eight zoospores. J Mol Biol. 1967;25(1):47–66.
Suto Y, Ohtani S. Strigula smaragdula complex (Lichenized Ascomycota, Strigulaceae) on living leaves of woody plants from Shimane-ken, Western Japan. Lichenology. 2011;10(1):1–13.
Grung M, D’Souza FML, Borowitzka M, Liaaen-Jensen S. Algal carotenoids 51. Secondary carotenoids 2. Haematococcus pluvialis aplanospores as a source of (3S, 3′S)-astaxanthin esters. J Appl Phycol. 1992;4(2):165–71.
Saini RK, Shetty NP, Prakash M, Giridhar P. Effect of dehydration methods on retention of carotenoids, tocopherols, ascorbic acid and antioxidant activity in Moringa oleifera leaves and preparation of a RTE product. J Food Sci Technol. 2014;51(9):2176–82.
Hellebust JA, Ahmad I. Regulation of nitrogen assimilation in green microalgae. Biol Oceanogr. 1989;6(3–4):241–55.
Kumar A, Bera S. Revisiting nitrogen utilization in algae: A review on the process of regulation and assimilation. Bioresour Technol Rep. 2020;12:100584.
Silva NFP, Gonçalves AL, Moreira FC, Silva TFCV, Martins FG, Alvim-Ferraz MCM, et al. Towards sustainable microalgal biomass production by phycoremediation of a synthetic wastewater: A kinetic study. Algal Res. 2015;11:350–8.
Giordano M. Interactions between C and N metabolism in Dunaliella salina cells cultured at elevated CO2 and high N concentrations. J Plant Physiol. 2001;158(5):577–81.
Almahrouqi H, Naqqiuddin MA, Achankunju J, Omar H, Ismail A. Different salinity effects on the mass cultivation of Spirulina (Arthrospira platensis) under sheltered outdoor conditions in Oman and Malaysia. J Algal Biomass Util. 2015;6:1–14.
Moronta R, Mora R, Morales E, Moronta R, Mora R, Morales E. Response of the microalga Chlorella sorokiniana to pH, salinity and temperature in axenic and non axenic conditions. Rev Fac Agron Univ Zulia. 2006;23(1):28–43.
Ambati RR, Gogisetty D, Aswathanarayana RG, Ravi S, Bikkina PN, Bo L, et al. Industrial potential of carotenoid pigments from microalgae: Current trends and future prospects. Crit Rev Food Sci Nutr. 2019;59(12):1880–902.
Panahi Y, Yari Khosroushahi A, Sahebkar A, Heidari HR. Impact of cultivation condition and media content on chlorella vulgaris composition. Adv Pharm Bull. 2019;9(2):182–94.
Li X, Li W, Zhai J, Wei H. Effect of nitrogen limitation on biochemical composition and photosynthetic performance for fed-batch mixotrophic cultivation of microalga Spirulina platensis. Bioresour Technol. 2018;263:555–61.
Li X, Li W, Zhai J, Wei H, Wang Q. Effect of ammonium nitrogen on microalgal growth, biochemical composition and photosynthetic performance in mixotrophic cultivation. Bioresour Technol. 2019;273:368–76.
Chowdury KH, Nahar N, Deb UK. The growth factors involved in microalgae cultivation for biofuel production: a review. Comput Water Energy Environ Eng. 2020;9(4):185–215.
Zhang YM, Chen H, He CL, Wang Q. Nitrogen starvation induced oxidative stress in an oil-producing green alga Chlorella sorokiniana C3. PLOS ONE. 2013;8(7):e69225.
Wu M, Zhu R, Lu J, Lei A, Zhu H, Hu Z, et al. Effects of different abiotic stresses on carotenoid and fatty acid metabolism in the green microalga Dunaliella salina Y6. Ann Microbiol. 2020;70(1):48.
Mulders KJM, Janssen JH, Martens DE, Wijffels RH, Lamers PP. Effect of biomass concentration on secondary carotenoids and triacylglycerol (TAG) accumulation in nitrogen-depleted Chlorella zofingiensis. Algal Res. 2014;6(Part A):8–16.
Urreta I, Ikaran Z, Janices I, Ibañez E, Castro-Puyana M, Castañón S, et al. Revalorization of Neochloris oleoabundans biomass as source of biodiesel by concurrent production of lipids and carotenoids. Algal Res. 2014;5:16–22.
Del Campo JA, García-González M, Guerrero MG. Outdoor cultivation of microalgae for carotenoid production: current state and perspectives. Appl Microbiol Biotechnol. 2007;74(6):1163–74.
Pelah D, Sintov A, Cohen E. The effect of salt stress on the production of canthaxanthin and astaxanthin by Chlorella zofingiensis grown under limited light intensity. World J Microbiol Biotechnol. 2004;20(5):483–6.
Sun XM, Ren LJ, Zhao QY, Ji XJ, Huang H. Microalgae for the production of lipid and carotenoids: a review with focus on stress regulation and adaptation. Biotechnol Biofuels. 2018;11(1):272.
Chen M, Tang H, Ma H, Holland TC, Ng KYS, Salley SO. Effect of nutrients on growth and lipid accumulation in the green algae Dunaliella tertiolecta. Bioresour Technol. 2011;102(2):1649–55.
Begum R, Howlader S, Mamun-Or-Rashid ANM, Rafiquzzaman SM, Ashraf GM, Albadrani GM, et al. Antioxidant and signal-modulating effects of brown seaweed-derived compounds against oxidative stress-associated pathology. Oxid Med Cell Longev. 2021;2021:e9974890.
Pereira AG, Otero P, Echave J, Carreira-Casais A, Chamorro F, Collazo N, et al. Xanthophylls from the Sea: algae as source of bioactive carotenoids. Mar Drugs. 2021;19(4):188.
Kokabi M, Yousefzadi, Yousefzadi M, Ahmadi A, Feghhi, Amin M, et al. Antioxidant activity of extracts of selected algae from the Persian Gulf, Iran. J Persian Gulf. 2013;4:45–50.