Isolation and characterization of bioactive protein from several Species of sponges as antibacterial agents Harningsih Karim (a,c) Ahyar Ahmad (b*), Nur Asmi (b), Muh. Nasrum Massi (c), Andi Ilham Latunra (d)
a) Department of Pharmacy, School of Pharmacy YAMASI, Makassar, Indonesia.
b) Chemistry Department, Mathematics and Natural Science Faculty, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea, Makassar 90245, Indonesia.
*ahyarahmad[at]gmail.com
c) Department of Medicine, Faculty of Medicine, Hasanuddin University, Jl Perintis Kemerdekaan Km. 10 Tamalanrea, Makassar 90245, Indonesia.
d) Biology Department, Mathematics and Natural Science Faculty, Hasanuddin University, Jl. Perintis Kemerdekaan Km. 10 Tamalanrea, Makassar 90245, Indonesia.
Abstract
Research on antibacterial bioactivity of protein fraction derived from various species of sponges in Barang Lompo island, South Sulawesi by using a polar solvent buffer of 0.1 M Tris-HCl with pH 8.3, 2 M NaCl, 0.01 M CaCl2, 1 % β-mercaptoethanol, 0.5 % Triton X-100 has been carried out. Lowry protein assay was performed to determine the protein concentration. The bioactivity test was analyzed using gel diffusion technique. Results have shown that the protein concentration in the crude extracts isolated from 500 g of fresh samples of four species of sponges (BLS 02, BLS 06, BLS 07, and BRL 01) was 7.080 mg/mL; 8.400 mg/mL; 16.624 mg/mL, respectively. The initial purification of protein use was carried out using the fractionation method with ammonium sulfate, followed by the dialysis process. Bioactivity against to Salmonella typhi was shown by all protein fractions, of which protein fraction with saturation level of ammonium sulfate 40-60% of BRL 01 sponge shows the highest inhibition zone (26.48 mm) followed by protein fraction with ammonium sulfate saturation level of 30-40% of BLS 06 sponge (26.18 mm).Test of the inhibition level in the various concentration of bioactive protein with the highest inhibition zone showed that the maximum activity was in 4000 μg/mL of protein from BRL 01 40-60 % and BLS 04 30-40 % with inhibition zones of 23.54 and 20.56 mm, respectively. These results indicate that bioactive protein from the sponge has great potential as a raw material for developing novel antibacterial drugs, particularly for Salmonella typhi infections.
Keywords: Bioactivity, sponges, bioactive protein, antibacterial, inhibition zone
