ANALYSIS OF BIOACTIVE COMPOUNDS AND ANTIMICROBIAL ACTIVITY OF MARINE ALGAE KAPPAPHYCUS ALVAREZII USING THREE SOLVENT EXTRACTS

ANALYSIS OF BIOACTIVE COMPOUNDS AND ANTIMICROBIAL ACTIVITY OF MARINE ALGAE KAPPAPHYCUS ALVAREZII USING THREE SOLVENT EXTRACTS

V. Prabha*¹, D.J. Prakash ² and P.N. Sudha³

Department of Biochemistry, D.K.M College for Women ¹, Vellore-632 001, Tamil Nadu, India

Neurophysiology Laboratory P.G. & Research Department of Zoology ², Voorhees College, Vellore-632 001, Tamil Nadu, India

P.G. and Research Department of Chemistry, D.K.M College for Women ³, Vellore-632 001, Tamil Nadu, India

ABSTRACT

The seaweeds are economically valuable resources, used as food, fodder, fertilizer and medicine and thus useful to mankind in many ways. In the present study, Kappaphycus alvarezii, a marine alga, has been analysed for the presence of bioactive products using three solvent extracts. Antimicrobial activity was also done using the same extracts of seaweed. The results revealed that the selected seaweed has active secondary metabolites and also exhibited antimicrobial activity, mainly in the methanolic extract of Kappaphycus alvarezii.

Seaweeds, Kappaphycus alvarezii, Secondary metabolites, Antimicrobial activity

INTRODUCTION: Economical significance of seaweed lies in their utilization in food and cosmetic industry and also in medicine and pharmacy. Genera of agar containing red seaweed, Gelidium Sp. and genera of carrageenan-containing red seaweed. Kappaphycus alvarezii are used most widely ¹.

The chemical composition of seaweed provides their high nutritional value contributing to human nutrients-such as proteins with all essential amino acids, minerals and vitamins.

In addition, they consist of bioactive secondary metabolites and many different compounds with health benefits ^{2, 3, 4}.

The chemical composition of seaweed varies, which is dependent on the type of species, habitat, time of collection, external conditions such as temperature, light and nutrient concentration in water ^{5, 6}. Bioactivity of diverse secondary metabolites and other compounds extracted from seaweeds plays an important role in prevention of different serious diseases. These biogenic compounds, also have antibacterial, antialgal, antifungal properties ^{7, 8}.

The antimicrobial activity of seaweeds are associated to secondary metabolites like terepenoid derivatives, phlorotannins or phenolic lipids ^{9, 10}.

Thus, the present study aims at the extraction and qualitative detection of various phytochemicals of biological importance, from the selected seaweed Kappaphycus alvarezii using three solvent extracts. Antimicrobial activity was also done using the same solvent extracts of the seaweed.

MATERIALS AND METHODS:

Collection and Identification of Seaweed:              The sea weed Kappaphycus alvarezii was purchased from the Mandabam region, Tamil Nadu, India and identified by SNAP natural and alginate products pvt ltd, Sipcot Ranipet, Tamil Nadu, India.

Preparation of Plant Extracts: The seaweed was washed thoroughly to remove epiphytes and other debris.  Then they were shade dried and coarse powdered. The powder obtained (250gms) were extracted successively with 3 solvents, Ethanol, Methonol and Acetone in a soxhlet extractor for 18-20hrs.

All those extracts was concentrated at 45^oC under reduced pressure using rotary flash evaporator.  All the three extracts were used for phytochemical screening and the methanolic extract was used for antimicrobial studies.

1. Phytochemical Screening:         All the extracts of seaweed (Kappaphycus alvarezii) were subjected to various qualitative tests for the identification of phytochemicals such as alkaloids, carbohydrates, glycosides, proteins and aminoacids, phytosterols, phenolic compounds, flavanoids, terepenoids, tannins and saponins, by using standard methods ¹¹.
2. Antimicrobial studies:
3. Preparation of Extract For Microbial Studies: For microbiological studies, 1 gm of dried methanol extract was dissolved in 10 ml of the same solvent.
4. Test Organisms Used: The bacterial and the fungal strains that were used for the determination of antimicrobial activity of the selected seaweed are as follows,
5. Bacterial strains - Staphylococcus aureus, Micrococcus lutus, Klebsiella pneumonia, E.Coli, Pseudomonas aeruginosa, Bacillus cereus.
6. Fungal strains - Aspergillus flavus, Aspergillus niger, Aspergillus fumigates, Candida albicans, Candida tropicals.

Well Diffusion Method: Antimicrobial assay was done using Muller Hinton Agar (MHA) and Potato Dextrose Agar (PDA). Sterilized medium was poured into a petridish and was inoculated by streaking the swabs of the test organisms. After allowing the inoculums to dry at room temperature, methanol extract of Kappaphycus alvarezii was then loaded in the well bored in solidified agar. The plates were allowed to stand at room temperature for 1 hour for extract to diffuse into the agar and then they were incubated at 37⁰ for 24 hours.  After incubation, the zone of inhibition exhibited by the extract was measured.

RESULTS AND DISCUSSION:

Phytochemical Screeing of Kappaphycus alvarezii extracts: Table 1 demonstrates qualitative phyto-chemical screening of ethanol, methanol and acetone extracts of Kappaphycus alvarezii. The results revealed the presence of various phytochemical constituents in all the extracts used. The methanolic extract showed a maximum number of active phytochemicals compared to ethanolic and acetone extracts.

TABLE 1: QUALITATIVE PHYTOCHEMICAL SCREENING OF VARIOUS EXTRACTS OF KAPPAPHYCUS ALVAREZII

+ = Presence ;      - = Absent

Antimicrobial activity of Methanolic Extract of Kappaphycus alvarezii: Antimicrobial activity of seaweed is mainly due to the presence of phenolic lipids, terepenes and phlorotannins. Since antimicrobial agents required for the antimicrobial activity were maximum present in the methanolic extract of the selected seaweed. The antimicrobial assay was done using the methanolic extract of the seaweed⁹ (Table 2). The zone of inhibition was observed. The selected seaweed exhibited antimicrobial activity for 3 bacteria and 3 fungi out of 6 and 5 test micro-organisms used respectively (Fig. 1 and Fig. 2).

TABLE 2: ANTIMICROBIAL ACTIVITY OF METHANOLIC EXTRACT OF KAPPAPHYCUS ALVAREZII

C – CONTROL ; K – KAPPAPHYCUS ALVAREZII

FIG. 1: ANTIBACTERIAL ACTIVITY OF METHANOLIC EXTRACT OF KAPPAPHYCUS ALVAREZII

C – CONTROL ; K – KAPPAPHYCUS ALVAREZII

Fig. 2: Antifungal activity of methanolic extract of Kappaphycus alvarezii

In summary, of the 3 solvent extracts of the seaweed, methanolic extract showed potential antibacterial activity ¹² against gram positive and gram negative organisms, and antifungal activities against selected fungal species. Also results of phytochemical screening showed the presence of carbohydrates, proteins, sterols, phenolic compounds, tannins, terepenes etc, in the solvents used, and which act as potential antimicrobial agent, when extracted with methanol.

CONCLUSION: The result of this study justifies that the marine algae, Kappaphycus alvarezii is associated with numerous health benefits with all its bioactive compounds, and can be recommended for the development of antimicrobial drugs.

ACKNOWLEDGMENT: We are grateful to the Department of Biochemistry, D.K.M. College for Women, Thiruvalluvar University, Tamil Nadu, India, for providing necessary facilities for carrying out this work.

REFERENCES:

1. MC Hugh, D.J. A guide to the seaweed industry, FAO fisheries technical paper 441. Food and Agriculture organization of the United Nations, Rome, 2003.
2. Cardozo, K.H.M., Guaratini, T., Barros, M.P. Metabolites from Algae with economical Impact. Comp. Biochem. Physiol., Part C, 2007, 146, 60-78.
3. Cho, Y.S., Jung, W.K., Kim, J.A., Choi, I.W. and Kim, S.K. Beneficial effects of Fucoidan on Osteoblastic MG-63 cell differentiation. Food Chem., 2009, 116, 990-994.
4. Artan, M., Li, Y., Karadeniz, F., Lee, S.H., Kim, M.M. and Kim, S.K. Anti-HIV-1 activity of phloroglucinol derivate, 6,6-bieckol, from ecklonia cava. Bioorg. Med. Chem.,2008, 16, 7921-7926.
5. Mabeau, S. and Fleurence, J. Seaweed in food products: biochemical and nutritional aspects. Trends Food Sci. technol., 1993, 4, 103-107.
6. Marinho-Soriano, E., Fonseca, P.C., Carneiro, M.A.A and Moreira, W.S.C. Seasonal variation in the chemical composition of two tropical seaweeds. Biores. Technol., 2006, 97, 2402-2406.
7. Paul V.J. and Puglish M.P. Chemical mediation of interactions among marine organisms. Nat. Prod. Rep., 2004, 21, 189-209.
8. Bhadury P. and Wright P.C. Exploitation of marine algae: Biogenic compounds for potential antifouling applications. Planta, 2004, 219, 561-578.
9. Sastry, V.M.V.S and Rao, G.R.K. Antibacterial substance from marine algae: successive extraction using benzene, chloroform and methanol. Bot. Mar.,1994,  37, 357-360.
10. Van Heemst J.D.H., Peuleve S. and De Leeuw J.W. Novel algal polyphenolic bio macromolecules as significant contributors to resistant fractions of marine dissolved and particulate organic matter. Org. Geochem., 1996, 24 (6-7), 629-640.
11. Kokate CK. Practical pharmacognosy; fourth edition; reprint 2001; Vallabh Prakasan, Pune. 2003, p.149-156.
12. Vairappan C.S., Suzuki M., Abe T. and Masuda M. Halogenated metabolites with antibacterial activity from the okinawan laurencia species. Phytochemistry, 2001, 58, 517-523.
    

    ---

    How to cite this article:

    Prabha V, Prakash DJ and Sudha PN: Analysis of Bioactive compounds and Antimicrobial activity of Marine algae Kappaphycus alvarezii using Three Solvent Extracts. Int J Pharm Sci Res. 2013; 4(1); 306-310.

    ---