شناسایی ترکیبات زیست فعال موجود در عصاره جلبک‌های قهوه‏ای سارگاسوم (Sargassum angustifulium) و پادینا (Padina distromatic) و بررسی اثرات ضد میکروبی، آنتی ‏اکسیدانی و خواص آنزیمی آن‌ها

نوع مقاله : مقاله پژوهشی

نویسندگان

• خدیجه شیرانی بیدابادی ¹
• شیلا صفائیان ¹
• رضوان موسوی ندوشن ¹
• ناهید رحیمی فرد ²

¹ گروه صنایع غذایی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران

² دانشکده پیراپزشکی، دانشگاه علوم پزشکی ایران، تهران، ایران

https://doi.org/10.22067/ifstrj.2021.73113.1104

چکیده

جلبک­های دریایی جز منابع غنی گیاهی محسوب می­شوند که در طب سنتی کاربردهای گوناگونی دارند. هدف از این مطالعه عصاره گیری دو گونه جلبک پادینا و سارگاسوم به دو روش ماسیراسیون و اولتراسوند به همراه آنالیز ترکیبات آن­ها و بررسی اثر آنتی اکسیدانی، ضد میکروبی و خواص آنزیمی عصاره­ها بوده است. نتایج حاکی از آن است که بازده عصاره­گیری جلبک سارگاسوم به روش ماسیراسیون از باقی عصاره‏ها بالاتر بوده است (09/3 g/100g) و عصاره اولتراسوندی این جلبک خواص آنتی‏اکسیدانی و ترکیبات فنولی و فلاونوئیدی بالاتری داشته است. مهار آنزیم استیل کولین استراز و میزان فعالیت آنزیم نیتریک اکساید در عصاره اولتراسوندی پادینا در بالاترین مقدار بود.. به طور کلی در همه عصاره­ها باکتری‏های Escherichia coli و  Listeria innocuaاز بقیه میکروارگانیسم‏ها مقاوم تر بودند و نتایج آزمون میکروبی نشان داد عصاره اولتراسوند پادینا از باقی عصاره‏ها به طور معناداری خاصیت ضد میکروبی بیشتری داشته است. با توجه به نتایج به دست آمده روش اولتراسوند روش مناسبی جهت عصاره‏گیری می­باشد و همچنین از این عصاره می­توان به عنوان ترکیب آنتی‏اکسیدان، ضد میکروب، ضد آلزایمر و کاهش‏دهنده نیترات در افزودنی‏های غذایی استفاده کرد.

کلیدواژه‌ها

• جلبک
• GC-MS
• خواص آنزیمی
• عصاره‌گیری
• ضد میکروبی

موضوعات

• زیست فناوری مواد غذایی

عنوان مقاله [English]

Identification of Bioactive Compounds in the Extracts of Brown Algae Sargassum (Sargassum angustifulium) and Padina )Padina distromatic( and Evaluation of Antimicrobial, Antioxidant and Enzymatic Properties

نویسندگان [English]

• Khadijeh Shirani Bidabadi ¹
• Shilla Safaeian ¹
• Rezvan Mousavi Nadushan ¹
• Nahid Rahimifard ²

¹ Department of Food Science and Technology, Faculty of Biological Science, North Tehran Branch, Islamic Azad University, Tehran, Iran

² Faculty of Paramedicine, Iran University of Medical Science, Tehran, Iran

چکیده [English]

Introduction
Sargassum and Padina are two genera of brown algae that are widely scattered in temperate regions. Sargassum species are categorized as tropical and sub-tropical brown seaweed which are valuable sources of bioactive compounds including dietary fibers, carotenoids, vitamins, and minerals. These brown algae demonstrate diverse biological activities such as antioxidant, antimicrobial, and anti‑Alzheimer, due to the presence of flavonoids, triterpenoids, flavonoids, sterols, polyphenols, and pheophytine. The genus Padina is scattered in many environmental conditions, mainly in the tropical marine waters, and belongs to the family Dictyotaceae. Some bioactive components isolated from Padina species have been demonstrated hypoglycemic, hypolipidemic, anti-obesity, hepatoprotective, cardioprotective, immunostimulatory, and antimicrobial activities., The aim of this study was to prepare an extract from two species of algae Padina and Sargsum by massaging and ultrasound assisted-methods  as well as analyzing their compounds and investigating the antioxidant, antimicrobial and enzymatic properties of the extracts. According to the obtained results, ultrasound assisted method was a suitable method for extraction. This extract can be used as a combination of antioxidant, antimicrobial, anti-Alzheimer's and nitrate reducing agent in food additives.
 
Materials and Methods
Chemical materials were supplied by Sigma-Aldrich GmbH (Sternheim, Germany). The algal species utilized in the current investigation; namely, Padina distromatic and Sargassum angustifolium were collected from the coastal region of Chabahar bandar, Sistan and Baluchistan Province, Iran. To eliminate all the impurities and extraneous materials, they were washed by using distilled water and then dried at ambient temperature (24-48 h) until the constant weight. Extraction by maceration was compared with the extraction using ultrasonic assisted method. Determination of chemical compounds was parformed using GC-MS device. Investigation of antioxidant properties and total polyphenol and flavonoid content were also performed. The degree of free radical scavenging was done according to DPPH method. Evaluation of antimicrobial effect of algae extracts were the main challenges in our research. S. aureus ATCC25923, Listeria innocua ATCC 33090, E. coli ATCC 25922 and S. typhi ATCC 6539 were used for antimicrobial test. Determination of minimum growth inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were performed using wells in agar.The enzymatic activity was also determined. In this study, the activity of acetylcholinesterase was investigated using the method introduced by Ellman et al. (1961) and also the activity of nitric oxidase was determined using a kit protocol (Nvand-Iran). Factorial experiment in the form of a completely randomized design was used to analyze the data. Duncan's multiple range test was used to determine the difference between the means at the 95% confidence level, and SAS software version 2.9 will used for statistical analysis.
 
Results and Discussion
The current study investigated the antioxidant, antibacterial, anti-inflammatory, and anti-Alzheimer’s attributes of two brown algae namely Padina distromatic and Sargassum angustifolium which were collected from the coastal region of Chabahar Bandar, Iran. The results clearly indicated that the type of algae and extraction techniques used in this investigation highly affected phytochemical compositions, antioxidant, AChE inhibitory, scavenging, and antimicrobial activities. Considering both extraction yield and phytochemical components, extraction by ultrasound assisted method provided better results. Among all algal extracts, UPE presented the highest AChE inhibitory activity, and antibacterial activity and USE presented the highest antioxidant activities, total phenols and flavonoids, reflecting the presence of various bioactive components. The extracts of two various seaweeds utilized in the current study highlighted considerable inhibitory action against four pathogenic bacteria. According to the observations of the antibacterial assay, S. aureus was the most sensitive microorganism, while L. innocua was revealed as the most resistant bacteria to the extracts of P. distromatic and S. angustifolium. Further, the chemical components responsible for the antioxidant, AChE inhibitory, and antibacterial power were confirmed by GC-MS analysis. The findings of the current investigation confirmed the potential of the health benefits and therapeutic effect of brown marine algae. Thus, the extract of P. distromatic and S. latifolium could be an effective supplement to be incorporated into the products’ formulation in the food and pharmaceutical industries as well as in medication to alleviate several disorders such as Alzheimer.

کلیدواژه‌ها [English]

• Algae
• Antimicrobial
• Enzymatic properties
• Extraction
• GC-MS

مراجع

1. Cox, S., Abu-Ghannam, N., & Gupta, S. (2010). An assessment of the antioxidant and antimicrobial activity of six species of edible Irish seaweeds. International Food Research Journal, 17(1), 205-220. https://doi.org/10.21427/D7HC92.
2. Duarte, M.E., Cardoso, M.A., Noseda, M.D., & Cerezo, A.S. (2001). Structural studies on fucoidans from the brown seaweed Sargassum stenophyllum. Carbohydrate Research, 333(4), 281-293. https://doi.org/10.1016/S0008-6215(01)00149-5.
3. Yende, S.R., Harle, U.N., & Chaugule, B.B. (2014). Therapeutic potential and health benefits of Sargassum Pharmacognosy Reviews, 8(15), 1. https://doi.org/10.4103/0973-7847.125514.
4. Latifi, A. M., Sadegh Nejad, E., & Babavalian, H. (2015). Comparison of extraction different methods of sodium alginate from brown alga Sargassum localized in the Southern of Iran. Journal of Applied Biotechnology Reports, 2(2), 251-255.
5. Amini, F., Riahi, H., & Zolgharnain, H. (2013). Metal concentrations in Padina species and associated sediment from Nayband Bay and Bostaneh Port, northern coast of the Persian Gulf, Iran.
6. Rohani-Ghadikolaei, K., Abdulalian, E., & Ng, W.K. (2012). Evaluation of the proximate, fatty acid and mineral composition of representative green, brown and red seaweeds from the Persian Gulf of Iran as potential food and feed resources. Journal of Food Science and Technology, 49(6), 774-780. https://doi.org/10.1007/s13197-010-0220-0.
7. Devi, K.N., Kumar, T.T., & Balasubramanian, T. (2014). Antibacterial and antioxidant effects from seaweed, Sargassum wightii (Greville, 1848) against marine ornamental fish pathogens. Journal of Coastal Life Medicine, 2(10), 773-83.
8. Kordjazi, M., Etemadian, Y., Shabanpour, B., & Pourashouri, P. (2019). Chemical composition antioxidant and antimicrobial activities of fucoidan extracted from two species of brown seaweeds (Sargassum ilicifolium and angustifolium) around Qeshm Island. Iranian Journal of Fisheries Sciences, 18(3), 457-475. https://doi.org/10.22092/IJFS.2018.11549.
9. Hlila, M.B., Hichri, A.O., Mahjoub, M.A., Mighri, Z., & Mastouri, M. (2017). Antioxidant and antimicrobial activities of Padina pavonica and Enteromorpha from the Tunisian Mediterranean coast. Journal of Coastal Life Medicine, 5, 336-342.
10. Vieitez, I., Maceiras, L., Jachmanián, I., & Alborés, S. (2018). Antioxidant and antibacterial activity of different extracts from herbs obtained by maceration or supercritical technology. The Journal of Supercritical Fluids, 133, 58-64. https://doi.org/10.1016/j.supflu.2017.09.025.
11. Pezeshkpour, V., Khosravani, S.A., Ghaedi, M., Dashtian, K., Zare, F., Sharifi, A., & Zoladl, M. (2018). Ultrasound assisted extraction of phenolic acids from broccoli vegetable and using sonochemistry for preparation of MOF-5 nanocubes: Comparative study based on micro-dilution broth and plate count method for synergism antibacterial effect. Ultrasonics Sonochemistry, 40, 1031-1038. https://doi.org/10.1016/j.ultsonch.2017.09.001.
12. Hameed, I.H., Hussein, H.J., Kareem, M.A., & Hamad, N.S. (2015). Identification of five newly described bioactive chemical compounds in methanolic extract of Mentha viridis by using gas chromatography-mass spectrometry (GC-MS). Journal of Pharmacognosy and Phytotherapy, 7(7), 107-125. https://doi.org/10.5897/JPP2015.0349.
13. Papandreou, M.A., Dimakopoulou, A., Linardaki, Z.I., Cordopatis, P., Klimis-Zacas, D., Margarity, M., & Lamari, F.N. (2009). Effect of a polyphenol-rich wild blueberry extract on cognitive performance of mice, brain antioxidant markers and acetylcholinesterase activity. Behavioural Brain Research, 198(2), 352-358. https://doi.org/10.1016/j.bbr.2008.11.013.
14. Chang, C.C., Yang, M.H., Wen, H.M., & Chern, J.C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of Food and Drug Analysis, 10(3). https://doi.org/10.38212/2224-6614.2748.
15. Brand-Williams, W., Cuvelier, M.E., & Berset, C.L.W.T. (1995). Use of a free radical method to evaluate antioxidant activity. LWT-Food science and Technology, 28(1), 25-30. https://doi.org/10.1016/S0023-6438(95)80008-5.
16. Ellman, G.L., Courtney, K.D., Andres Jr, V., & Featherstone, R.M. (1961). A new and rapid colorimetric determination of acetylcholinesterase activity. Biochemical Pharmacology, 7(2), 88-95. https://doi.org/10.1016/0006-2952(61)90145-9.
17. Behbahani, B.A., Yazdi, F.T., Vasiee, A., & Mortazavi, S.A. (2018). Oliveria decumbens essential oil: Chemical compositions and antimicrobial activity against the growth of some clinical and standard strains causing infection. Microbial Pathogenesis, 114: 449-452. https://doi.org/10.1016/j.micpath.2017.12.033.
18. Noshad, M., Hojjati, M., & Behbahani, B.A. (2018). Black Zira essential oil: Chemical compositions and antimicrobial activity against the growth of some pathogenic strain causing infection. Microbial Pathogenesis, 116, 153-157. https://doi.org/10.1016/j.micpath.2018.01.026.
19. Ferrazzano, G.F., Scioscia, E., Sateriale, D., Pastore, G., Colicchio, R., Pagliuca, C., & Pagliarulo, C. (2017). In vitro antibacterial activity of pomegranate juice and peel extracts on cariogenic bacteria. BioMed Research International. https://doi.org/10.1155/2017/2152749.
20. Abdel-Aal, E.I., Haroon, M., & Mofeed, J. (2015). “Successive solvent extraction and GC-MS analysis for the evaluation of the phytochemical constituents of the filamentous green Alga Spirogyra longata.” Egyptian Journal of Aquatic Research, 41(3), 233–46. https://doi.org/10.1016/j.ejar.2015.06.001.
21. Sarpal, A.S., & Costa, I.C.R. (2016). Investigation of Biodiesel Potential of Biomasses of Microalgaes Chlorella, Spirulina and Tetraselmis by NMR and GC-MS Techniques. Journal of Biotechnology & Biomaterials, 06(01), 1–15. https://doi.org/10.1016/j.ejar.2015.06.001.
22. Avula, S.G.C., Belovich, J.M., & Xu, Y. (2017). Determination of fatty acid methyl esters derived from Algae Scenedesmus dimorphus Biomass by GC–MS with one-step esterification of free fatty acids and transesterification of glycerolipids. Journal of Separation Science, 40(10), 2214–27. https://doi.org/10.1002/jssc.201601336
23. Barbalace, M. C., Malaguti, Giusti, L., Lucacchini, A., Hrelia, S., & Angeloni, C. (2019). Anti-inflammatory activities of marine algae in neurodegenerative diseases. International Journal of Molecular Sciences, 20(12), https://doi.org/10.3390/ijms20123061.
24. Boi, V.N., Dang, X. C., & Phan, T. K. V. (2016). Effects of extraction conditions over the Phlorotannin content and antioxidant activity of extract from brown algae Sargassum serratum (Nguyen Huu Dai 2004). Free Radicals and Antioxidants, 7(1), 115–22. https://doi.org/10.5530/fra.2017.1.17
25. Chemat, F., Rombaut, N., Sicaire, A.G., Meullemiestre, A., Fabiano-Tixier, A.S., & Abert-Vian, M. (2017). Ultrasound assisted extraction of food and natural products. Mechanisms, Techniques, Combinations, Protocols and Applications. A Review.” Ultrasonics Sonochemistry, 34, 540–60. https://doi.org/10.1016/j.ultsonch.2016.06.035.
26. Franco, D., Sineiro, J., Rubilar, M., Sánchez, Jerez, M., Pinelo, M., Costoya, N., & Núñez, M. J. (2008). Polyphenols from plant materials: extraction and antioxidant power. Electronic Journal of Environmental, Agricultural and Food Chemistry, 7(8), 3210–16.
27. Fujii, T., Y. Gao, R., Sharma, E.L., Hu, DenBaars, S.P., & Nakamura, S. (2004). Increase in the extraction efficiency of GaN-based light-emitting diodes via surface roughening. Applied Physics Letters, 84(6), 855–57. https://doi.org/10.1063/1.1645992
28. Gaysinski, M., Ortalo-Magné, A., Thomas O.P., & Culioli, G. (2015). Springer Protocols, Natural Products from Marine Algae, 2, 207-223. https://doi.org/10.1007/978-1-4939-2684-8-13.
29. Hielscher-Michael, S., Griehl, C., Buchholz, M., Ulrich Demuth, H., Arnold, N., & Ludger, A.W. (2016). Natural products from microalgae with potential against Alzheimer’s disease: Sulfolipids Are Potent Glutaminyl Cyclase inhibitors. Marine Drugs, 14(11), 203. https://doi.org/10.3390/md14110203.
30. Hlila, M.B., Majouli, K., Jannet, H.B., Mahjoub, A., Mastouri, M., & Selmi, B. (2017). Antimicrobial activity of Tunisian Euphorbia paralias Asian Pacific Journal of Tropical Biomedicine, 7(7), 629–32. https://doi.org/10.1016/j.apjtb.2017.06.008.
31. Kausalya, M., & Narasimha Rao, G. M. (2015). Antimicrobial activity of marine algae. Journal of Algal Biomass Utilization, 6(1), 78–87.
32. Lin, H.W., Liu, C.W., Yang, D.J., Chen, C.C., Shih Yin, C., Jung Kai T., Tien Jye C., & Yuan Yen, C. (2017). Dunaliella salina alga extract inhibits the production of Interleukin-6, Nitric Oxide, and Reactive Oxygen species by regulating nuclear factor-ΚB/Janus Kinase/Signal Transducer and activator of transcription in Virus-infected RAW264.7 Cells. Journal of Food and Drug Analysis, 25(4), 908–18. https://doi.org/10.1016/j.jfda.2016.11.018
33. Machu, L., Misurcova, L., Ambrozova, J.V., Orsavova, J., Mlcek, J., Sochor, J., & Jurikova, T. (2015). Phenolic content and antioxidant capacity in Algal food products. Molecules, 20(1), 1118–33. https://doi.org/10.3390/molecules20011118
34. Mekinić, I. G., Skroza, D., Šimat, V., Hamed, I., Čagalj, M., & Perković, Z.P. (2019). Phenolic content of brown algae (Pheophyceae) species: extraction, identification, and quantification. Biomolecules, 9(6), 244. https://doi.org/10.3390/biom9060244.
35. Moein, S., Moein, M., Ebrahimi, N., Farmani, F., Sohrabipour, J., & Rabiei, R. (2015). Extraction and determination of protein content and antioxidant properties of ten algae from Persian Gulf. International Journal of Aquatic Science, 6(2), 29–38.
36. Olasehinde, T. A., Olaniran, A. O., & Okoh, A. I. (2019). Macroalgae as a valuable source of naturally occurring bioactive compounds for the treatment of Alzheimer’s disease. Marine Drugs, 17(11), 609. https://doi.org/10.3390/md17110609.
37. Olasehinde, T.A., Olaniran, A.O., Okoh, A.I., & Koulen, P. (2017). Therapeutic potentials of microalgae in the treatment of Alzheimer’s disease. Molecules, 22(3), 480. https://doi.org/10.3390/molecules22030480.
38. Otero, P., Quintana, S. E., Reglero, G., Fornari, T., & García-Risco, R. M. (2018). Pressurized liquid extraction (PLE) as an innovative green technology for the effective enrichment of Galician algae extracts with high quality fatty acids and antimicrobial and antioxidant properties. Marine Drugs, 16(5), 156. https://doi.org/10.3390/md16050156.
39. Patil, P.D., Dandamudi, K.P.R., Wang, J., Deng, Q., & Deng, S. (2018). Extraction of bio-oils from Algae with supercritical carbon dioxide and co-solvents. Journal of Supercritical Fluids, 135, 60–68. https://doi.org/10.1016/j.supflu.2017.12.019.
40. Rathnayake, A.U., Abuine, R., Kim, Y.J., & Byun, H.G. (2019). Anti-Alzheimer’s materials isolated from marine bio-resources: a review. Current Alzheimer Research, 16(10), 895–906. https://doi.org/10.2174/1567205016666191024144044.
41. Silva, A., Silva, S.A., Carpena, M., Garcia-Oliveira, P., Gullón, P., Barroso, M.F., Prieto, M.A., & Simal-Gandara, J. (2020). Macroalgae as a source of valuable antimicrobial compounds: extraction and applications. Antibiotics, 9(10), 1–41. https://doi.org/10.3390/antibiotics9100642.
42. Uysal, S., Aktumsek, A., Picot, C.M.N., Sahan, A., Mollica, A., Zengin, G., & Mahomoodally, M.F. (2017). A comparative: in vitro and in silico study of the biological potential and chemical fingerprints of Dorcycinum pentapyllum Haussknechtii using three extraction procedures. New Journal of Chemistry, 41(22), 13952–60. https://doi.org/10.1039/C7NJ03497K.
43. Varshney, P., Beardall, J., Bhattacharya, S., & Wangikar, P.P. (2018). Isolation and biochemical characterisation of two thermophilic green algal species- asterarcys Quadricellulare and Chlorella Sorokiniana, which are tolerant to high levels of carbon dioxide and nitric oxide. Algal Research, 30, 28–37. https://doi.org/10.1016/j.algal.2017.12.006.
44. Wen, C., Zhang, J., Zhang, H., Dzah, C. S., Zandile, M., Duan, Y., Ma, H., & Luo, X. (2018). Advances in ultrasound assisted extraction of bioactive compounds from cash crops – a review. Ultrasonics Sonochemistry, 48(May), 538–49. https://doi.org/10.1016/j.ultsonch.2018.07.018.