DETERMINATION OF SOME BIOACTIVITIES OF Convallaria majalis L. (LILY OF THE VALLEY), ISOLATION PHARMACEUTICAL ACTIVE INGREDIENT AND INVESTIGATION ITS INDUSTRIAL USEAGE

Year 2022, Volume: 8 Issue: 2, 122 - 137, 31.12.2022

Nazan Demir , Sıla Nezahat Daşdemir , Alevcan Kaplan , Yaşar Demir

https://doi.org/10.51477/mejs.1196088

Cited By: 1

Abstract

In this study, some biological activities of Convallaria majalis L. (Asparagaceae), which grows naturally in Muğla and its surroundings and attracts attention with its pleasant smell, were determined, and the isolation of drug active substances and industrial usability were investigated. For this purpose, the protease enzyme that catalyzes the hydrolysis of proteins, which is one of the most important enzyme groups in both industrial and biochemical applications, into peptides and amino acids was purified from C. majalis. Protease enzyme was purified using TPP (Three Phase Partitioning) method. Optimum pH and optimum temperature for the enzyme, Km and Vmax values for casein, azocasein, gelatin, hemoglobin, azoalbumin substrates were determined. SDS-PAGE was used to check the purity of the purified protease enzyme. The molecular weight of the enzyme was calculated as 54.347 kDa using gel filtration chromatography. The effects of SDS, EDTA, β-mercaptoethanol compounds and Mg2+, Ca2+, Mn2+, Ni2+, Hg2+, Fe2+, Fe3+ cations, 10 mM, 1 mM, 0.1 mM concentrations on enzyme activity were investigated. Volatile and flavoring components analyzed with Headspace GC/MSD. It was determined that C.majalis flowers contain volatile organic compounds, citronellol (9.6%), geraniol (8.4%), benzyalcohol (35%), phenylacetonitrile (3.0%), farnesol (1.9 %) and 2,3-dihydrofarnesol (0.88 %), green grassy notes: (Z)-3-hexen-1-ol (11 %), (Z)-3-hexenyl acetate (7.8%). As a result of the work, it was determined that the aromatic of C. majalis can be used as an active ingredient and it has been concluded that there is an industrial use.

Keywords

: C.majalis, Pharmaceuticals, Protease Enzyme, TPP, Volatile and Flavoring Components

Supporting Institution

Muğla Sıtkı Koçman Üniversitesi

Project Number

15/236

Thanks

This research subject was supported by the Scientific Research Projects Coordination Unit of Muğla Sıtkı Koçman University with the project numbered 15/236. The authors thank Muğla Sıtkı Koçman University Scientific Research Projects Coordination Unit for their support.

References

• [1] Daşdemir, SN., “Müge Çiçeğinin (Convallaria majalis) Bazı Biyoaktivitelerinin Belirlenmesi Ve Diğer Bazı Çiçeklerle Birlikte Parfüm Tasarımında Kullanılabilirliğinin Araştırılması”, Yüksek Lisans Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü,Muğla, (2017).
• [2] Khanin, V.A., Moiseev, O.O., Babkina, А.Y., Kotenko, O.M., Kotov, A.G., “Development of the New Approach to Quality Control of Extracts of the Plant Raw Material on the Example of Convallaria majalis”, News of Pharmacy, 2,86,2016.
• [3] Godswill, A.C., “Medicinal Plants: The Medical, Food, and Nutritional Bıichemistry and uses”, International Journal of Advanced Academic Research | Sciences, Technology and Engineering, 5, 11,220-241, 2019.
• [4] Faydaoğlu, E., Sürücüoğlu, M., “Geçmişten Günümüze Tıbbi ve Aromatik Bitkilerin Kullanılması ve Ekonomik Önemi”, Kastamonu Üniversitesi Orman Fakültesi Dergisi, 11,1, 52-67, 2011.
• [5] Davis, P.H., “Flora of Turkey and the East Aegean Islands”,Vol.8, Edinburgh at the University Press, pp.84, 1984.
• [6] Yıldırım, H., “Asparagaceae Familyası Altında Yer Alan Bazı Cinslere (Scila, Puschkinia, Chionodoxa, Prospero, Muscari ve Bellevalia) Yönelik Sistematik Bir Yaklaşım”, Herbarium Turcicum, 1:1-14, 2022.
• [7] Davis, P.H., Mill, R.R., Tan, K., “Flora of Turkey and the east Aegean islands (Supplement)”, Edinburg Univ. Press, Edinburgh, 1988.
• [8 ] Güner, A., Aslan, S., (eds.) “Türkiye bitkileri listesi:(damarlı bitkiler)”, Nezahat Gökyiǧit Botanik Bahçesi Yayınları, 2012.
• [9] İlçim, A., Karataş, H., Karahan, F., “Bazı Muscari Mill. (Asparagaceae) Türleri Üzerine Karşılaştırmalı Morfolojik, Anatomik ve Palinolojik Çalışmalar”, Iğdır Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 10(2) 846-854, 2020.
• [10] Baytop, T., “Türkiye’de Bitkiler İle Tedavi”, Baskı II, Nobel Tıp Kitabevleri LTD., İstanbul, s. 234, 1999.
• [11] Chushenko, V.N., Vinnik , E.V., Komissarenko, N.F., Stupakova, E.P., Petukhova, T.V., Zinchenko, V.V., “Carbohydrates of the flowers of Convallaria majalis and C. keiskei”, Chemistry of Natural Compounds, 28, 242-243,1992.
• [12] Kopp, B., Kubelka, W., “New cardenolides from Convallaria majalis”, Planta Medica, 45,195-202, 1982.
• [13] Rawdkuen, S., Chaiwut, P., Pintathang, P., Benjakul, S., “Three-phase partitioning of protease from Calotropis procera latex”, Biochemical Engineering Journal, 50, 145-149,2010.
• [14] Fadıloğlu, S., “Immobilization and characterization of ficin”, Nahrung/Food, 45 (2), 143-146, 2001.
• [15] Laemmli, U.K.,“Cleavage of structural proteins during the assembly of the head of bacteriophage T4”. Nature, 227(5259),680-685,1970.
• [16] Bradford, M,M., “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding”, Analytical Biochemistry, 72, 248-254, 1976.
• [17] Leaman, D. J., “Sustainable Wild Collection of Medicinal and Aromatic Plants, Development of an International Standard”, Medicinal and Aromatic Plants, chapter 7, 97-107, 2006.
• [18] Ersöz, T., “Bitkisel Tedaviye Bilimsel Bakış: Doğrular ve Yanlışlar”. Journal of Pediatric Infection, 5, 217-222, 2011.
• [19] Özgen, Ö., Emiroğlu, H., Yıldız, M., Tag, A.S., Purutçuoğlu, E., “Tüketiciler ve Modern Biyoteknoloji: Model Yaklaşımlar”, Ankara Üniversitesi Biyoteknnoloji Enstitüsü Yayınları No,1, Ankara,4, 2007.
• [20] González-Rábade, N., Badillo-Corona, J.A., Aranda-Barradas, J.S., Oliver Salvador, M.C., “Production of plant proteases in vivo and in vitro -A review. Biotechnology Advances, 29, 983-996, 2011.
• [21] Narayan, A.V., Madhusudhan, M.C., Raghavarao, K.S.M.S., “Extraction and purification of ipomoea peroxidase employing three-phase partitioning”, Applied Biochemistry and Biotechnology, 151, 263-272, 2008.
• [22] Singh, R.K., Gourinath, S., Sharma, S., Roy, I., Gupta, M.N., Betzel, C., Srinivasan, A., Singh, T. P., “Enhancement of enzyme activity through three-phase partitioning: Crystal structure of a modified serine proteinase at 1.5 Å resolution”, Protein Engineering, 14, 307-313,2001.
• [23] Uçkaya, F., “Citrus sinensis (L.) Osbeck (Portakal) ve Citrus limon (L.) Burm.F. (Limon) Çiçeklerinden Proteaz ve Peroksidaz Enzimlerinin Saflaştırılması ve Endüstride Kullanımlarının Araştırılması”, Doktora Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü,Muğla, 2015.
• [24] Gul, A., Khan, S., Arain, H., Khan, H., Ishrat, U., Siddiqui, M., “Three-phase partitioning as an efficient one-step method for the extraction and purification of bromelain from pineapple crown waste”, Journal of Food Processing and Preservation, 00,1-9, 2022.
• [25] Huan, XW., Chen, L.J., Luo, Y.B., Guo, H.Y., Ren, F.Z., “Purification, characterization, and milk coagulating properties of ginger proteases”, Journal of Dairy Science, 94, 2259-2269,2011.
• [26] Nafi, A., Ling, F,H., Bakar, J., Ghazali, H.M., “Partial characterization of an enzymatic extract from Bentong ginger (Zingiber officinale var. bentong)”, Molecules, 19, 12336-12348, 2014.
• [27] Demir, Y., Güngör, A.A., Duran, E.D., Demir, N., “Cysteine Protease (Capparin) from Capsules of Caper (Capparis spinosa)”, Food Technology and Biotechnology, 46 (3), 286-291,2008.
• [28] Demir, N., Taşğın, E., “Hindiba (Cichorium intybus L.) Bitkisinden Myrosinaz Enziminin Saflaştırılması, Karakterize Edilmesi ve Kozmetik Alanında Kullanılabilirliğinin İncelenmesi”, Süleyman Demirel Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 16, 307-314, 2012.
• [29] Demir, N., Daşdemir, S.N., Can, Z., “Purification and Characterization of Protease Enzyme from Oleander (Nerium oleander) Flowers of Different Colors”, International Journal of Innovative Research and Reviews,1(1),21-26, 2017.
• [30] Febriani, K., Wahyuni, I., Setiasih, S., Hudiyono, S., “Comparative study of two methods of fractionation bromelain from pineapple core extract (Ananas comosus)”, In AIP Conference Proceedings, 1862 (1), 030095, AIP Publishing LLC, 2017.
• [31] Banik, S., Biswas, S., Karmakar, S., “Extraction, purification, and activity of protease from the leaves of Moringa oleifera [version 1; referees: 2 approved, 1 approved with reservations]” F1000Research, 7,1151, 2018.
• [32] Khan, A.A., Fazili, A.B.A., Bhat, S.A., Bhat, W.F., Asghar, M.N., Khan, M.S., Bano, B., “Purification, characterization and studies of a novel cysteine protease inhibitor from Juglans regia: Implications as a potential biopesticide”, Journal of King Saud University – Science, 34, 101829, 2022.
• [33] Demiray, A., “Kozmetik Amaçlı Kullanım İçin Kır Menekşesinde (Viola odorata) Bazı Biyokimyasal Aktivitelerin Aranması Ve Tanımlanması”, Yüksek Lisans Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü, Muğla, 2013.
• [34] Akçay, O., “Kızılçam Kabuğundan (Pinus brutia) Etken Madde İzolasyonu ve Değişik Enzim Aktiviteleri Üzerindeki Etkilerinin İncelenmesi”, Yüksek Lisans Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü, Muğla, 2014.
• [35] Demirkent, B.P., “Datça Papatyasının (Chrysanthemum coronarium) Bazı Biyoaktif Bileşenlerinin Belirlenmesi; İlaç ve Kozmetik Endüstrisinde Hammadde Olarak Kullanılabilirliğinin Araştırılması”, Yüksek Lisans Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü, Muğla, 2015.
• [36] Işık, C., “Mis Zambağının (Lilium candidum) Biyokimyasal Aktivitelerinin Belirlenmesi ve endüstriyel Amaçlı Kullanılabilirliğinin Araştırılması”, Yüksek Lisans Tezi, Muğla Sıtkı Koçman Üniversitesi, Fen Bilimleri Enstitüsü, Muğla, 2014.
• [37] Yıldırım Çelik, S., “Çiğdem (Crocus biflorus) Yumrularından Proteaz Enziminin Saflaştırılması ve Saflaştırılan Enzimin Kazeinin Koagülasyonunda Kullanılabilirliğinin Araştırılması”, Gıda, 43 (2), 231-239, 2018.
• [38] Alıcı, E.A, “Çilek Meyvesinden Proteaz Enziminin Saflaştırılması, Karakterizasyonu ve Uygulama Alanlarının Araştırılması”, Doktora Tezi, Sakarya Üniversitesi, Fen Bilimleri Enstitüsü, Sakarya, 2019.
• [39] Lehninger, A. L., “Principles of biochemistry”, Worth Publishers Inc., New York, pp, 1152, 2013.
• [40] Liu, L., Zhu, Y., Chen, Y., Chen, H., Fan, C., Mo, Q., Yuan, J., “One-Pot Cascad Biotransformation for Efficient Synthesis of Benzyl Alcohol and Its Analogs”, Chemistry-An Asian Journal, 15, 101, 2020.
• [41] Nie, C., Xiao Du, Y.G., Bian, J.L., Li H., Zhang, X., Wang, C.M., Li, S.Y., “Characterization of the efect of cis‑3‑hexen‑1‑ol on green tea aroma”, Scientific Reports. 10: 15506, 2020.
• [42] Brito, R.G., Guimarães, A.G., Quintans, J.S.S., Santos, M.R.V., De Sousa, D.P, Jr Badaue-Passos, D., de Lucca Jr, W., Brito, F.A., Barreto, E.O., Oliveira, A.P., Quintans, L.J., “Citronellol, a monoterpene alcohol, reduces nociceptive and inflammatory activities in rodents”, Journal of Natural Medicines, 66, 637-644,2012.
• [43] Pereira de Lira, M.H., de Andrade Júniorb, F.P., Queiroga Moraesc, G.F., da Silva Macenac, G., de Oliveira Pereira F., Lima, I.O., “Antimicrobial activity of geraniol: an integrative review”, Journal of Essential Oil Research, 32 (3), 187-197, 2020.