Evaluation of biological activities and chemical composition in extracts of red algae Laurencia (Rhodomelaceae, Ceramiales) from the coast os Espírito Santo, Brazil

Author(s)

Erika Mattos Stein

• Document type: Master's Dissertation
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Biociências (IBIOC/SB)
• Defense date: 2011-06-21

Examining board members

Mutue Toyota Fujii; Maria Luiza Faria Salatino; Valeria Laneuville Teixeira

• Advisor: Mutue Toyota Fujii

Abstract

The Red Algae, phylum Rhodophyta, represent one of the largest and oldest lineages of eukaryotic organism. From the chemical standpoint, the Laurencia J.V. Lamouroux (Ceramiales) has been identified as one of the most complete and diverse genus within this specific group. The members of this genus have been identified, for instance, as the largest producers of secondary metabolites known to the Rhodophyta. Thus, it is not surprising that these specific species are currently under intense scrutiny with regard to their potential as new sources of natural products for medical and biotechnological applications. The primary aim of this study is to start a systematic investigation on the identification and characterization of novel Laurencia natural products showing desirable pharmacological and biomedical properties. The species used in this research are L. aldingensis (LA), L. catarinensis (LC), L. dendroidea(LD), L. intricate (LI). In addition, L. translucida (LT), Palisada flagellifera(PF), P.perforata (PP) and a pigment variant of LD (LDV) were included for comparison. Specifically, algal extracts were obtained through the sequential fractionation of dried algae samples with hexane (EH), chloroform (EC), methanol (EM), water (EA), and a single, crude, aqueous extracts (EAB). Protein content was performed using the Bradford, bicinchoninic acid assay (BCA) and the 280 nm uv quantification methods. The Bradford method was identified to be more appropriate for quantification. The analysis of phycobilins revealed that phycoerythrin was the major pigment in all species tested. Extracts were assayed for antimicrobial activity following the NCCLS microdilution tests and the minimum inhibitory concentration (MICs) obtained spectrophotometrically. Antimicrobial activity was explored using a variety of model biological targets (i.e. pathogens). To this end, microdilutions were used following the classical NCCLS and a minimal inhibitory concentration (MIC) protocol was performed on basis of electronic spectroscopy measurements. With respect to our findings on antibacterial activity, the LA-EH extract was bactericidal against the P. aeruginosa concentration of 62.8 μg.mL-1 extracts and LA-EM and LC-EH had a strong bacteriostatic action against S. pneumoniae at concentrations of 51.1 and 85,1 μg.mL-1, respectively. In the antifungal assay, LA-EH and LA-EC were fungicides against C. parapsilosis to 49.0 and 57.8 μg.mL-1, respectively. The antioxidant assay was tested using the method with DPPH, in which the EC presented them as possessing the finest extracts of molecules capable of donating protons. And for the anticholinesterase test was done in the qualitative assay and quantitative assay on TLC plate by the Ellman method that could demonstrate the inhibitory activity of AChE less than 50 % for concentrations of 600 μg.mL-1. The profile was similar for all extracts in which the EH and EC were the most active and possibly EM gave a false positive, while the EA and EAB extracts had very low activity. In the trial against the pathogen Colletotrichum Gloesporioides that causes papaya anthracnose the results show that LC-EH and LD-EH are the most active with IC50 of 70 and 40 μg.mL-1, respectively. To perform the cytotoxicity assay against mammalian cells a model that is under development was proposed in which use cells MES-SA and its corresponding mutant (MES-SA/Dx5) multi-drug resistant. The proposed model is quite promising and the initial screening we propose a more detailed investigation of LD-EH and LT-EH whose IC50 were 91 μg.mL-1 and 16 μg.mL-1, respectively, against MES-SA cells. The LA, LC, LD and LI extract compounds were analyzed by GC-MS and the identification was performed using the library data and literature by comparing the mass spectra, retention time and index Kovat\'s. The study conducted here is likely to draw directions and help in the search for new molecules or group of molecules capable of being used in therapy without the toxicity of chemically synthesized compounds. (AU)

• FAPESP's process: 08/07775-0 - Evaluation of Biological activities of extracts from red algae Laurencia (Rhodomelaceae, Ceramiales)
• Grantee: Erika Mattos Stein
• Support Opportunities: Scholarships in Brazil - Master