Sys Rev Pharm 2020;11(9):518-521 A multifaceted review journal in the field of pharmacy 518 Systematic Reviews in Pharmacy Vol 11, Issue 9, Sep-Oct 2020 Protein Isolation from Sponge Niphates sp. as an Antibacterial and Antioxidant Warsidah 1 , Masrianih 2 , Mega Sari Juane Sofiana 1 , Ikha Safitri 1 , Ajuk Sapar 3 , Anthoni B. Aritonang 3 , Yuges Saputri Muttalib 4 , Dzul Fadly 5 * 1 Department of Marine Science, Faculty of Math and Natural Science, Tanjungpura University, Pontianak, Indonesia 2 Department of Biology Education, Faculty of Teacher Training and Education, Tadulako University, Palu City, Indonesia 3 Department of Chemistry, Faculty of Math and Natural Science, Tanjungpura University, Pontianak, Indonesia 4 Department of Nutrition, Faculty of Health Sciences, Esa Unggul University, Jakarta, Indonesia 5 Department of Food Technology, Faculty of Agriculture, Tanjungpura University, Pontianak, Indonesia * Corresponding author: Dzul Fadly. Email : dzul.fadly@faperta.untan.ac.id ABSTRACT Research on the antioxidant and antibacterial activity of the isolated protein of Niphates sp. sponge origin Spermonde aquatic of South Sulawesi had been conducted. Determination of antimicrobial activity was based on the formation of inhibition zones/cleared areas surrounding the isolated protein for two tested bacteria, i.e., Bacillus subtilis and Escherichia coli. The isolated protein from this sponge has the highest antimicrobial activity in E. coli bacterial assay by 14.21 mm zone of inhibition, while the inhibition zone of B. subtilis was 12.05 mm. After extra time for 24 hours of incubation, the medium of B. subtilis showed turbidity in the clear spot before. This indicated that the isolated protein covered the bacteriostatic ability against the tested bacteria. The antioxidant activity assay by DPPH free radical scavenging effect of isolated protein with some levels of saturation was determined. Following the test results indicated the antioxidant activity of isolated protein with scavenging effect against free radical DPPH possessed IC50 values from crude protein extract of 5.05 μg/mL, protein fraction 0 – 30% of 203.71 μg/mL, protein fraction 31 – 50% of 163.75 μg/mL, protein fraction 51 – 70% of 111.31 μg/mL, and protein fraction 71 – 90% amounted to 590.03 μg/mL. The results indicated that the sponge material's protein was potential as an antioxidant and antimicrobial, which can then be further research to several other biologic activities. Keywords: Niphates sp., Bacillus subtilis, Escherichia coli, Antioxidant, Antibacterial Correspondence: Dzul Fadly Department of Food Technology, Faculty of Agriculture, Tanjungpura University, Pontianak, Indonesia Email : dzul.fadly@faperta.untan.ac.id INTRODUCTION Sponge in the waters is one of the reservoir animals for marine microbes that reached 60% of sponge's total biomass. The microbial symbionts produce active compounds to respond to extreme environmental conditions through the body's defense mechanisms. The sponge can associate with many different microorganisms such as cyanobacteria, heterotrophic bacteria (Hentschel et al., 2002). Sea sponge has a densely of microorganisms potentially benefitted as a pharmacological activity (Hentschel et al., 2001). The sponge and bacteria interact with commensalism symbiosis, which produces bioactive compounds (Roy et al., 2000). Microbial metabolites associate with marine invertebrates has structural similarities to their host (Proksch et al., 2002; Putri et al., 2015; Thiel and Imhoff, 2003). Some of the activities shown by microbes associated sponges, among others, were Microascus strain K14 and Monochaetia strain 193A20 shows antimicrobial activity. Penicilliumbrocae associated sponge Zyzzya sp. shows antimicrobial activity against methicillin-resistant Staphylococcus aureus. The cytotoxic activity was demonstrated by fungi Gymnascela dankaliensis associated sponge Halichondria japonica (Bugni and Ireland, 2004). Some of the compounds produced from the sponges, especially those in Indonesia's waters, among others were compound sesquiterpenoids type bisabolen, curcuphenol, and curcudiol resulting from Axynissa sp. have an activity inhibiting the synthesis of protein kinase on testing the anticancer in vitro (Hertiani et al., 2008). Also, Barrangamida compound, a new cyclic polypeptide, is active compounds produced by Theonella swinhoei sponge origin Spermonde waters of South Sulawesi (Roy et al., 2000). Cytotoxic activity against cervical cancer of Cinachyrella sp. sponge origin of the Situbondo coast has been reported (Nurhayati et al., 2015). Niphates sp. sponge is one of the sponges growing in Spermonde waters South Sulawesi. There are no reports or the scientific study of this sponge species' activity, and it becomes the basis for testing some of the biological activity of this sponge protein. The antimicrobial and antioxidant activity of a sponge protein is an early indication of the potential natural product for further testing, such as anticancer and some other important biological activities. This study was an effort to determine the antimicrobial and antioxidant activity of Niphates sp. sponge protein origin Spermonde waters of South Sulawesi. MATERIALS AND METHODS Sample preparation Samples taken from the sponge reefs in Spermonde waters of South Sulawesi was cleaned, then milled while added buffer A (0.1 M Tris-HCl pH 8.3, 2 M NaCl, 0.01 M CaCl2, 1% B-mercaptoethanol, Triton X-100 0.5%) and then stored in the refrigerator at 4 °C overnight). The suspension of Niphates sp. was conducted through the filtration using the Buchner funnel. After, the filtrate was freeze and thawed for 2-3 times. Next, it was centrifuged 30 min. at 6000 rpm and 4 °C, then the obtained supernatant kept in a refrigerator before subjecting to further analysis. Fractionation with ammonium sulfate Protein crude extract was fractionated by using ammonium sulfate at saturation levels (0-90%) (Bollag et al., 1996). Ammonium sulfate precipitation process performed by the saturation level of 0-30%; 30-50%; 50-70%, and 70-90%. Protein crude extracts in which certain volume was added with ammonium sulfate at a certain level of saturation while stirring with a magnetic stirrer until completely