Evaluation of the anti-bacterial activity of methanolic extract of Chlorella vulgaris Beyerinck [Beijerinck] with special reference to antioxidant modulation

Biswajita Pradhan1, Srimanta Patra1, Soumya Ranjan Dash1, Rabindra Nayak1, Chhandashree Behera1, Mrutyunjay Jena1
1Algal Biotechnology and Molecular Systematic Laboratory, Post Graduate Department of Botany, Berhampur University, Bhanja Bihar, Berhampur 760007, India

Tóm tắt

Abstract Background The natural antioxidants from Chlorella have potent therapeutic implication in several diseases. However, the anti-bacterial activity and their molecular mode of action have not been investigated yet. The present study focussed on the assessment of antioxidant potential as well as free radical scavenging activity such as DPPH, hydroxyl radical, hydrogen peroxide, and superoxide anion radical assay of Chlorella vulgaris Beyerinck [Beijerinck] (BUACC25) isolated from marine habitat. Furthermore, the anti-bacterial activity and their molecular mode of action have been evaluated. Results In the present study, the preliminary phytochemical screening of methanolic algal extract revealed the presence of alkaloids, glycosides, proteins, terpenoids, saponins, coumarin, phenols, and tannins, which was confirmed by in an UV-visible and FT-IR spectroscopy, indicated the distinct spectral peaks. The methanolic algal extract was found to be rich in phenolic content (45 ± 0.06 mg GAE g−1) and flavonoid content (470 ± 0.25 mg of RUE g−1). Furthermore, the methanolic extract was revealed potent antioxidant scavenging activity to scavenge various free radicals with minimum IC50 values of DPPH, hydroxyl, H2O2, superoxide 2.82 ± 0.30, 2.30 ± 0.25, 3.24 ± 0.32, and 3.15 ± 0.02 μg ml−1 respectively. Furthermore, the methanolic extract of C. vulgaris exhibited potent anti-bacterial activity which was evident with the reduction in cfu × 107/ml and % of cell viability. Mechanistically, reduction of SOD, CAT, and GSH activity provoked ROS-mediated cell death after drug treatment. Moreover, in combination with norfloxacin and ciprofloxacin, methanolic extract of C. vulgaris demonstrated enhanced anti-bacterial activity with an evident reduction in cfu/ml and % of cell viability. Conclusion This study advocates that C. vulgaris (BUACC25) has promising antioxidant activity owing to the presence of phenolic and flavonoids evidenced by scavenging of DPPH, hydroxyl, H2O2, and superoxide radicals. In addition to this, it sustained anti-microbial activity against E. coli through modulation of SOD, CAT, and GSH. This study carved a path for uncovering a better therapeutic agent against disease-causing bacterial pathogens.

Từ khóa


Tài liệu tham khảo

Maharana S, Pradhan B, Jena M, Misra MK (2019) Diversity of phytoplankton in Chilika Lagoon, Odisha, India. Environ Ecol 37(3):737–746

Malve H (2016) Exploring the ocean for new drug developments: marine pharmacology. J Pharm Bioallied Sci 8(2):83–91. https://doi.org/10.4103/0975-7406.171700

Patra S, Praharaj PP, Panigrahi DP, Panda B, Bhol CS, Mahapatra KK, Mishra SR, Behera BP, Jena M, Sethi G, Patil S, Patra SK, Bhutia SK (2020) Bioactive compounds from marine invertebrates as potent anticancer drugs: the possible pharmacophores modulating cell death pathways. Mol Biol Rep. https://doi.org/10.1007/s11033-020-05709-8

Wells ML, Potin P, Craigie JS, Raven JA, Merchant SS, Helliwell KE, Smith AG, Camire ME, Brawley SH (2017) Algae as nutritional and functional food sources: revisiting our understanding. J Appl Phycol 29(2):949–982. https://doi.org/10.1007/s10811-016-0974-5

Pradhan B, Patra S, Nayak R, Behera C, Dash SR, Nayak S, Sahu BB, Bhutia SK, Jena M (2020) Multifunctional role of fucoidan, sulfated polysaccharides in human health and disease: A journey under the sea in pursuit of potent therapeutic agents. Int J Biol Macromol. https://doi.org/10.1016/j.ijbiomac.2020.09.019

Gam DH, Yi Kim S, Kim JW (2020) Optimization of ultrasound-assisted extraction condition for phenolic compounds, antioxidant activity, and epigallocatechin gallate in lipid-extracted microalgae. Molecules 25(3). https://doi.org/10.3390/molecules25030454

Stabili L, Acquaviva MI, Angile F, Cavallo RA, Cecere E, Del Coco L, Fanizzi FP, Gerardi C, Narracci M, Petrocelli A (2019) Screening of chaetomorpha linum lipidic extract as a new potential source of bioactive compounds. Mar Drugs 17(6). https://doi.org/10.3390/md17060313

Olasehinde TA, Olaniran AO, Okoh AI (2019) Phenolic composition, antioxidant activity, anticholinesterase potential and modulatory effects of aqueous extracts of some seaweeds on beta-amyloid aggregation and disaggregation. Pharm Biol 57(1):460–469. https://doi.org/10.1080/13880209.2019.1634741

Narasimhan MK, Pavithra SK, Krishnan V, Chandrasekaran M (2013) In vitro analysis of antioxidant, antimicrobial and antiproliferative activity of enteromorpha antenna, enteromorpha linza and gracilaria corticata extracts. Jundishapur J Nat Pharm Prod 8(4):151–159. https://doi.org/10.17795/jjnpp-11277

Chakraborty K, Maneesh A, Makkar F (2017) Antioxidant activity of brown seaweeds. J Aquatic Food Prod Technol 26(4):406–419. https://doi.org/10.1080/10498850.2016.1201711

Lauritano C, Ferrante MI (2019) Marine natural products from microalgae: an -omics overview. 17(5). doi:https://doi.org/10.3390/md17050269

Pradhan B, Patra S, Dash SR, Maharana S, Behera C, Jena M (2020) Antioxidant responses against aluminum metal stress in Geitlerinema amphibium. SN Appl Sci 2(5):800. https://doi.org/10.1007/s42452-020-2599-1

Rastogi RP, Singh SP, Hader DP, Sinha RP (2010) Detection of reactive oxygen species (ROS) by the oxidant-sensing probe 2',7'-dichlorodihydrofluorescein diacetate in the cyanobacterium Anabaena variabilis PCC 7937. Biochem Biophys Res Commun 397(3):603–607. https://doi.org/10.1016/j.bbrc.2010.06.006

Pisoschi AM, Pop A (2015) The role of antioxidants in the chemistry of oxidative stress: A review. Eur J Med Chem 97:55–74. https://doi.org/10.1016/j.ejmech.2015.04.040

Nunomura A, Castellani RJ, Zhu X, Moreira PI, Perry G, Smith MA (2006) Involvement of oxidative stress in Alzheimer disease. J Neuropathol Exp Neurol 65(7):631–641. https://doi.org/10.1097/01.jnen.0000228136.58062.bf

Leopold JA (2015) Antioxidants and coronary artery disease: from pathophysiology to preventive therapy. Coron Artery Dis 26(2):176–183. https://doi.org/10.1097/mca.0000000000000187

Kasote DM, Katyare SS, Hegde MV, Bae H (2015) Significance of antioxidant potential of plants and its relevance to therapeutic applications. Int J Biol Sci 11(8):982–991. https://doi.org/10.7150/ijbs.12096

Sansone C, Brunet C (2019) Promises and challenges of microalgal antioxidant production. Antioxidants 8(7). https://doi.org/10.3390/antiox8070199

Poprac P, Jomova K, Simunkova M, Kollar V, Rhodes CJ, Valko M (2017) Targeting free radicals in oxidative stress-related human diseases. Trends Pharmacol Sci 38(7):592–607. https://doi.org/10.1016/j.tips.2017.04.005

Martins N, Barros L, Ferreira IC (2016) In vivo antioxidant activity of phenolic compounds: Facts and gaps. Trends Food Sci Technol 48:1–12

Xu DP, Li Y, Meng X, Zhou T, Zhou Y, Zheng J, Zhang JJ, Li HB (2017) Natural antioxidants in foods and medicinal plants: extraction, assessment and resources. Int J Mol Sci 18(1). https://doi.org/10.3390/ijms18010096

Tavakoli J, Brewer S, Jelyani A, Estakhr P (2017) Oxidative stability of olive oil during thermal process: effect of Pistacia khinjuk fruit oil. Int J Food Prop 20. doi:https://doi.org/10.1080/10942912.2017.1285787

Younes G, Rasoul-Amini S, Morowvat MH (2011) Algae for the production of SCP. Bioprocess Sci Technol:Nova Science Publishers, Inc: 163–184

Lin PY, Tsai CT, Chuang WL, Chao YH, Pan IH, Chen YK, Lin CC, Wang BY (2017) Chlorella sorokiniana induces mitochondrial-mediated apoptosis in human non-small cell lung cancer cells and inhibits xenograft tumor growth in vivo. BMC Complement Altern Med 17(1):88. https://doi.org/10.1186/s12906-017-1611-9

Syed S, Arasu A, Ponnuswamy I (2015) The uses of Chlorella vulgaris as antimicrobial agent and as a diet: the presence of bio-active compounds which caters the vitamins, minerals in general. Int J Bio Sci Bio Technol 7(1):185–190

Pradhan B, Baral S, Patra S, Behera C, Nayak R, MubarakAli D, Jena M (2020) Delineation of gamma irradiation (60Co) induced oxidative stress by decrypting antioxidants and biochemical responses of microalga, Chlorella sp. Biocatalys Agric Biotechnol 25:101595. https://doi.org/10.1016/j.bcab.2020.101595

Sofowora A (1996) Research on medicinal plants and traditional medicine in Africa. J Altern Complement Med 2(3):365–372. https://doi.org/10.1089/acm.1996.2.365

Mohanty S, Pradhan B, Patra S, Behera C, Nayak R, Jena M (2020) Screening for nutritive bioactive compounds in some algal strains isolated from coastal Odisha. J Adv Plant Sci 10(2):1–8

Patra S, Panda PK, Naik PP, Panigrahi DP, Praharaj PP, Bhol CS, Mahapatra KK, Padhi P, Jena M, Patil S (2020) Terminalia bellirica extract induces anticancer activity through modulation of apoptosis and autophagy in oral squamous cell carcinoma. Food Chem Toxicol 136:111073

Patra S, Bhol CS, Panigrahi DP, Praharaj PP, Pradhan B, Jena M, Bhutia SK (2020) Gamma irradiation promotes chemo-sensitization potential of gallic acid through attenuation of autophagic flux to trigger apoptosis in an NRF2 inactivation signalling pathway. Free Radical Biol Med 160:111–124. https://doi.org/10.1016/j.freeradbiomed.2020.06.022

Quettier-Deleu C, Gressier B, Vasseur J, Dine T, Brunet C, Luyckx M, Cazin M, Cazin JC, Bailleul F, Trotin F (2000) Phenolic compounds and antioxidant activities of buckwheat (Fagopyrum esculentum Moench) hulls and flour. J Ethnopharmacol 72(1-2):35–42. https://doi.org/10.1016/s0378-8741(00)00196-3

Prieto P, Pineda M, Aguilar M (1999) Spectrophotometric quantitation of antioxidant capacity through the formation of a phosphomolybdenum complex: specific application to the determination of vitamin E. Anal Biochem 269(2):337–341. https://doi.org/10.1006/abio.1999.4019

Pradhan B, Patra S, Maharana S, Behera C, Dash SR, Jena M (2020) Demarcating antioxidant response against aluminum induced oxidative stress in Westiellopsis prolifica Janet 1941.1-14. Int J Phytoremediation. https://doi.org/10.1080/15226514.2020.1807906

Pradhan B, Patra S, Behera C, Nayak R, Patil S, Bhutia SK, Jena M (2020) Enteromorpha compressa extract induces anticancer activity through apoptosis and autophagy in oral cancer. https://doi.org/10.1007/s11033-020-06010-4

Liu F, Ooi VE, Chang ST (1997) Free radical scavenging activities of mushroom polysaccharide extracts. Life Sci 60(10):763–771. https://doi.org/10.1016/s0024-3205(97)00004-0

Karley D, Shukla SK, Rao TS (2018) Isolation and characterization of culturable bacteria present in the spent nuclear fuel pool water. Environ Sci Pollut Res Int 25(21):20518–20526. https://doi.org/10.1007/s11356-017-0376-5

Sieuwerts S, de Bok FA, Mols E, de Vos WM, Vlieg JE (2008) A simple and fast method for determining colony forming units. Lett Appl Microbiol 47(4):275–278. https://doi.org/10.1111/j.1472-765X.2008.02417.x

Ren Y, Chow LM, Leung WW (2013) Cell culture using centrifugal microfluidic platform with demonstration on Pichia pastoris. Biomed Microdev 15(2):321–337. https://doi.org/10.1007/s10544-012-9735-7

Bhaduri S, Demchick PH (1983) Simple and rapid method for disruption of bacteria for protein studies. Appl Environ Microbiol 46(4):941–943. https://doi.org/10.1128/AEM.46.4.941-943.1983

Scott MD, Meshnick SR, Eaton JW (1987) Superoxide dismutase-rich bacteria. Paradoxical increase in oxidant toxicity. J Biol Chem 262(8):3640–3645

Schwartz CE, Krall J, Norton L, McKay K, Kay D, Lynch RE (1983) Catalase and superoxide dismutase in Escherichia coli. J Biol Chem 258(10):6277–6281

Chaudhuri D, Ghate NB, Deb S, Panja S, Sarkar R, Rout J, Mandal N (2014) Assessment of the phytochemical constituents and antioxidant activity of a bloom forming microalgae Euglena tuba. Biol Res 47(1):24

Behera SK (2018) Phytochemical screening and antioxidant properties of methanolic extract of root of Asparagus racemosus Linn. Int J Food Prop 21(1):2681–2688. https://doi.org/10.1080/10942912.2018.1560310

Balboa EM, Conde E, Moure A, Falqué E, Domínguez H (2013) In vitro antioxidant properties of crude extracts and compounds from brown algae. Food Chem 138(2-3):1764–1785

Widowati I, Zainuri M, Kusumaningrum HP, Susilowati R, Hardivillier Y, Leignel V, Bourgougnon N, Mouget JL (2017) Antioxidant activity of three microalgae Dunaliella salina, Tetraselmis chuii and Isochrysis galbana clone Tahiti. In: IOP Conference Series: Earth and Environmental Science, vol 1. IOP Publishing, Bristol, England, p 012067

Jerez-Martel I, García-Poza S, Rodríguez-Martel G, Rico M, Afonso-Olivares C, Gómez-Pinchetti JL (2017) Phenolic profile and antioxidant activity of crude extracts from microalgae and cyanobacteria strains. J Food Qual 2017:1-8.

Choudhary RK, Swarnkar PL (2011) Antioxidant activity of phenolic and flavonoid compounds in some medicinal plants of India. Nat Prod Res 25(11):1101–1109. https://doi.org/10.1080/14786419.2010.498372

Boulet J, Ducasse M-A, Cheynier V (2017) Ultraviolet spectroscopy study of phenolic substances and other major compounds in red wines: relationship between astringency and the concentration of phenolic substances: UV spectroscopy of red wine components. Aust J Grape Wine Res 23. https://doi.org/10.1111/ajgw.12265

Rajeswari R, Jeyaprakash K (2019) Bioactive potential analysis of brown seaweed Sargassum wightii using UV-VIS and FT-IR. J Drug Deliv Ther 9:150–153. https://doi.org/10.22270/jddt.v9i1.2199

Gülçin İ, Huyut Z, Elmastaş M, Aboul-Enein HY (2010) Radical scavenging and antioxidant activity of tannic acid. Arab J Chem 3(1):43–53

Chatzikonstantinou M, Kalliampakou A, Gatzogia M, Flemetakis E, Katharios P, Labrou NE (2017) Comparative analyses and evaluation of the cosmeceutical potential of selected Chlorella strains. J Appl Phycol 29(1):179–188

Ozcelik B, Lee J, Min D (2003) Effects of light, oxygen, and pH on the absorbance of 2, 2-diphenyl-1-picrylhydrazyl. J Food Sci 68(2):487–490

Chen C, You L-J, Abbasi AM, Fu X, Liu RH (2015) Optimization for ultrasound extraction of polysaccharides from mulberry fruits with antioxidant and hyperglycemic activity in vitro. Carbohydr Polym 130:122–132

Soare JR, Dinis TC, Cunha AP, Almeida L (1997) Antioxidant activities of some extracts of Thymus zygis. Free Radical Res 26(5):469–478

Wijk RV, Wijk E, Van P, Wiegant FA, Ives J (2008) Free radicals and low-level photon emission in human pathogenesis: state of the art

Ebrahimzadeh M, Nabavi S, Nabavi S (2009) Antioxidant activities of methanol extract of Sambucus ebulus L. flower. Pakistan J Biol Sci 12(5):447

Yen GC, Duh PD (1994) Scavenging effect of methanolic extracts of peanut hulls on free-radical and active-oxygen species. J Agric Food Chem 42(3):629–632

Gülçin İ, Topal F, Sarikaya SBÖ, Bursal E, Bilsel G, Gören AC (2011) Polyphenol contents and antioxidant properties of medlar (Mespilus germanica L). Rec Nat Prod 5(3):158

Parejo I, Viladomat F, Bastida J, Rosas-Romero A, Flerlage N, Burillo J, Codina C (2002) Comparison between the radical scavenging activity and antioxidant activity of six distilled and nondistilled Mediterranean herbs and aromatic plants. J Agric Food Chem 50(23):6882–6890

Agrawal MK (2016) Antimicrobial activity of Nostoc calcicola (Cyanobacteria) isolated from central India against human pathogens. Asian J Pharm 10(04):554-559.

Reuk-ngam N, Chimnoi N, Khunnawutmanotham N (2014) Antimicrobial activity of coronarin D and its synergistic potential with antibiotics. BioMed Res Int 2014:581985. https://doi.org/10.1155/2014/581985