Bactericidal Potentiality of Purified Terpenoid Extracts from the Selected Sea Weeds and its Mode of Action

Sabira Siraj Sumayya, Abdulhadeef Shereefa Lubaina, Kumaraswamy Murugan


Terpenoids are hydrocarbons involved in a variety of basic functions in plants such as growth, development and other physiological events. Terpenes and their associated molecules safe guard the organisms from pest, pathogen and herbivores. Similarly, therapeutically terpenoids function as antimicrobial agents against bacteria, fungi and viruses. The mechanism of bactericidal activities may be via inhibiting the synthesis of essential molecules like proteins, nucleic acids, cell-wall components, cell membrane derailment, bacterial DNA replication or inhibition of metabolic pathways. The crude methanolic extracts of the seaweeds were subjected to silica gel column chromatographic purification and eluted with different combinations of ethyl acetate: petroleum ether solvent systems. The eluted fractions were further subjected to thin layer chromatography and fractionated by GC-MS. The fractions obtained from Hypnea musciformis revealed the terpenoids such as eicosane, heneicosane, 2-pentadecnone, hexadecanoic acid methyl ester, n- hexadecanoic acid, hexadecanoic acid ethyl ester, heptadecanoic acid methyl ester, 11-octadecanoic acid metyl ester, whereas Kappapycus alvarezii  displayed hexadecane, eicosane, heptadecane, octadecane, heneicosane, tricosane, hexadecanoic acid, methyl ester and beta amyrin. Similarly, Gracillaria dura revealed hexadecanoic acid methyl ester, n- hexadecanoic acid, 11-octadecanoic acid and phytol. Subsequently, the bactericidal activities of the purified terpenoid extracts from the sea weeds were carried. Initially, the extracts were tested for their in vitro antibacterial activity against six bacterial strains such as three Gram-positive (Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis) and three Gram-negative bacteria (Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa) by disc diffusion method. The results revealed that the purified terpenoid extracts of G. dura exhibited significant bactericidal potentiality against S. mutans as compared to other strains. The zone of inhibition, MIC and MBC values narrate the efficacy of the purified terpenoid extract of the species. Remarkable leaching of metabolites like protein and DNA further substantiates the MIC and MBC results. Scanning electron microscopic observations such as clumping, irregularity of cells and ballooned walls reflect the possible membrane damage accounted in the cells by the terpenoid extracts. Further studies are planned to validate the above data by using molecular tools.


Bactericidal activity, Disc diffusion method, Sea weeds, GC-MS, Terpenoids, SEM.

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Christianson DW (2017) Structural and Chemical Biology of Terpenoid. Chemical Reviews 117(17):11570–11648.doi: 10.1021 /acs.chemrev.7b00287

Jiang Z, Kempinski C, Chappell J (2016) Extraction and Analysis of Terpenes/Terpenoids. Current Protocols Plant Biology 1(4):345–358.doi: 10.1002/cppb.20024.

Oussalah M, Caillet S, Saucier L, Lacroix M (2007) Inhibitory effects of selected plant essential oils on the growth of four pathogenic bacteria: E. coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus and Listeria monocytogenes. Food Control 18(6): 414–420.doi: 10.1016/j.foodcont.2005.11.009.

Barbieri R, Coppo E, Marchese A, Daglia M, Nabavi S (2016) Phytochemicals for human disease: An update on plant-derived compounds antibacterial activity. Microbiological Research 3(7): 196-214. doi: 10.1016/j.micres.2016.12.003.

Mahizan NA, Yang SK, Moo CL, et al (2019) Terpene derivatives as a potential agent against antimicrobial resistance (AMR) Pathogens. Molecules 24(14): 2631 2641. doi :10.3390/molecules24142631.

Wagner H, Ulrich-Merzenich G (2009) Synergy research: Approaching a new generation of phytopharmaceuticals. Phytomedicine 16(4): 97–110. doi: 10.1016/j.fitote.2010.11.016 •

Bajpai VK, Sharma A, Baek KH (2013) Antibacterial mode of action of Cudrania tricuspidata fruit essential oil,affecting membrane permeability and surface characteristics of food-borne pathogens. Food Control 32: 582–590. doi:10.3389/fmicb.2017.00552

Humphries RM, Ambler J, Mitchell SL, Castanheira M, Dingle T, Hindler JA, Koeth L, Seii K (2018) CLSI methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests. Journal of Clinical Microbiology 56(4): e01934-17. doi:10.1128/JCM.01934-1

Du W, Sun C, Liang Z, Han Y, Yu J (2012) Antibacterial activity of hypocrellin A against Staphylococcus aureus. World Journal of Microbiology and Biotechnology 28(3): 3151–3157. doi: 10.1007/s11274-012-1125-z.

Xu JG, Hu QP, Wang XD, Luo JY, Liu Y, Tian CR (2010) Changes in the main nutrients, phytochemicals, and antioxidant activity in yellow corngrain during maturation. J. Agriculture and Food Chemistry 58(7): 5751–5756. doi: 10.1021/jf100364k.

Kaya I, Yigit N, Benli M (2008) Antimicrobial activity of various extracts of Ocimum basilicum L. and observation of the inhibition effect on bacterial cells by use of scanning electron microscopy. African Journal of Traditional, Complementary and Alternative Medicines 5(4): 363-369 doi: 10.4314/ajtcam.v5i4.31291.

Rota MC, Herrera A, Martinez RM, Sotomayor JA, Jordan MJ (2008) Antimicrobial activity and chemical composition of Thymus vulgaris, Thymus zygis and Thymus hyemalis essential oils. Food Control 19(4): 681–687. doi: 10.1016/j.foodcont.2007.07.00

Weerakkody NS, Caffin N, Turner MS, Dykes GA (2010) In vitro antimicrobial activity of less-utilized spice and herb extracts against selected food-borne bacteria. Food Control 21(5): 1408–1414. doi:10.1016/j.foodcont.2010.04.014.

Aligiannis N, Kalpoutzakis E, Mitaku S, Chinou IB (2001) Composition and antimicrobial activity of the essential oils of two Origanum species. Journal of Agriculture and Food Chemistry 49(2): 4168–4170.doi:10.1021/jf001494m.

Menon AN, Padmakumari KP, Jayalekshmy A (2003) Essential oil composition of four major cultivars of black pepper (Piper nigrum L.). Journal of Essential Oil and Research 15(1): 155–157. doi:10.1080/10412905.2003.9712099

Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods- a review. International Journal of Food Microbiology 94(2):223–253. doi:10.1016/j.ijfoodmicro.2004.03.022

Burt SA, Reinders RD (2003). Antibacterial activity of selected plant essential oils against Escherichia coli O157:H7. Letters in Applied Microbiology 36: 162-167.doi: 10.1046/j.1472-765X.2003.01285.x.

Elumalai EK, Ramachandran M, Thirumalai T, Vinothkumar P (2011) Antibacterial activity of various leaf extracts of Merremia emarginata. Asian Pacific Journal of Tropical Biomedicine 1(5):406–408.doi:10.1016/S2221-1691(11)60089-0

Santos SAO, Martins C, Pereira C, Silvestre AJD, Rocha SM (2019) Current challenges and perspectives for the use of aqueous plant extracts in the management of bacterial infections: the case-study of Salmonella enterica. International Journal of Molecular Science 20(4): 940 -947.doi:10.3390/ijms20040940.

Gonelimali FD, Lin J, Miao W, Xuan J, Charles F, Chen M, Hatab SR (2018) Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms Frontiers in Microbiology 8(2): 1639 -1643.doi: 10.3389/fmicb.2018.01639.

Atef NM, Shanab SM, Negm SI, Abbas YA (2019) Evaluation of antimicrobial activity of some plant extracts against antibiotic susceptible and resistant bacterial strains causing wound infection. Bulletin of the Natural Research Center 43(5):1-5.doi: 10.1186/s42269-019-0184-9.

Goldy Saxena, Kalra SS (2011) Antimicrobial activity pattern of certain terpenoids. International Journal of Pharmaceutical and Biological Science 2(1) 87-91.doi:

Manandhar S, Luitel S, Dahal R (2019) In Vitro antimicrobial activity of some medicinal plants against human pathogenic bacteria. Journal of Tropical Medicine 54(2):1-7.doi:org/10.1155/2019/1895340

Hajrah N, Abdul WM, Sabir J, Saleh Al-Garni SM, Sabir M, Salim MA, El-hamidy, Saini KS, Bora RS (2018) Anti-bacterial activity of Ricinus communis L. against bacterial pathogens Escherichia coli and Klebsiella oxytoca as evaluated by Transmission electron microscopy, Biotechnology and Biotechnological Equipment 32(3): 686-691.doi: 10.1080/13102818.2018.1451778.

Kaya I, Yiğit N, Benli M (2008) Antimicrobial activity of various extracts of Ocimum basilicum l. and observation of the inhibition effect on bacterial cells by use of scanning electron microscopy African Journal of Traditional, Complementary and alternative medicine 5(4): 363 – 369.doi: 10.4314/ajtcam.v5i4.31291

Prabhat, Ajaybhan, Navneet, Chauhan A (2010) Evaluation of antimicrobial activity of six medicinal plants against dental pathogens. Report and Opinion 2: 36-42.

Adamczak A, O˙zarowski M, Karpi´nski TM (2020) Antibacterial activity of some flavonoids and organic acids widely distributed in plants Journal of Clinical Medicine (9): 1-17. doi: 10.3390/jcm9010109.

Chiavari-Frederico MO, Barbosa LN, Carvalho dos Santos I, Ratti da Silva G, Fernandes de Castro A, de Campos Bortolucci W, Barboza LN et al (2020) Antimicrobial activity of Asteraceae species against bacterial pathogens isolated from postmenopausal women. PLoS ONE. 15(1): e0227023.

Guimarães AC, Meireles LM, Lemos MF, Guimarães MCC, Endringer DC, Fronza M, Scherer R (2019) Antibacterial activity of terpenes and terpenoids present in essential oils Molecules 24(4): 2471-2484.doi: 10.3390/molecules24132471.

Zhang J, Ye K P, Zhang X, Pan DD, Sun YY, Cao JX (2017) Antibacterial Activity and Mechanism of Action of Black Pepper Essential Oil on Meat-Borne Escherichia coli. Frontier in Microbiology 7:2094-20110.doi: 10.3389/fmicb.2016.02094.

Mahizan NA, Yang S , Moo C, Song AA, Chong C, Abushelaibi A, Lim S, Lai K (2019) Terpene derivatives as a potential agent against antimicrobial resistance (AMR) pathogens. Molecules 24(5): 2631-2645.doi: 10.3390/molecules24142631.

Irfan MS, Ahmed, Sharma M (2014) Antimicrobial activity of terpenoids from Sphaeranthus indicus. L Asian Journal of Plant Science and Research. 4(1):1-6.


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