A recombinant hybrid of manganese dependent-superoxide dismutase of Staphylococcus equorum and S. saprophyticus has successfully been overexpressed in Escherichia coli BL21(DE3), purified, and characterized. The recombinant enzyme suffered from degradation and aggregation upon storage at -20 °C, but not at room temperature nor in cold. Chromatographic analysis in a size exclusion column suggested the occurrence of dimeric form, which has been reported to contribute in maintaining the stability of the enzyme. Effect of monovalent (Na(+), K(+)), divalent (Ca(2+), Mg(2+)), multivalent (Mn(2+/4+), Zn(2+/4+)) cations and anions (Cl(-), SO4 (2-)) to the enzyme stability or dimeric state depended on type of cation or anion, its concentration, and pH. However, tremendous effect was observed with 50 mM ZnSO4, in which thermostability of both the dimer and monomer was increased. Similar situation was not observed with MnSO4, and its presence was detrimental at 200 mM. Finally, chelating agent appeared to destabilize the dimer around neutral pH and dissociate it at basic pH. The monomer remained stable upon addition of ethylene diamine tetraacetic acid. Here we reported unique characteristics and stability of manganese dependent-superoxide dismutase from S. equorum/saprophyticus.
Peroxide enzymes are widely used as a reagent in many clinical diagnosis of analysis methods such as ELISA, immunoblotting, and immunochemistry. Water spinach (Ipomea aquatica Forssk) is a species of plant available abundantly in Indonesia, hence it is a potential source of peroxidase. In this study, the crude extract of water spinach leaves was known to contain peroxidase enzyme which was active toward hydrogen peroxide. A fraction of extract yielded from differential fractionation using ammonium sulfate at concentration 40-65% (F4) was known to be the most active. Characterization of specific activity of peroxidase of F4 for optimum pH and temperature was at pH 5 and 60°C. The Michaelis-Menten constant (Km) was 0,27 mg/mL and enzyme's catalytic rate (Vmax) was 2,16 mg/mL/min.
One strategy to improve oral bioavailability of hydrophobic drugs like black seed oil with formulate them into self nanoemulsifying drug delivery system (SNEDDS). The aim of this study is to develop black seed oil into liquid SNEDDS formulation to enhance oral bioavailability. The optimization of SNEDDS was carried out using various comparison between oil, surfactant and cosurfactant. The formulated SNEDDS were evaluated for transmittance percentage, emulsification time, dispersebility, robustness, thermodynamics stability and globul size determination. The best formulation of SNEDDS obtained from tween 80 as surfactan and PEG 400 as cosurfactant (2:1) with ratio of oil and mix surfactant-cosurfactant 1:8. The formulation of SNEDDS met the requirement of transmittance percentage (97.80%), emulsification time (35.00 second), grade A of dispersibility, stable of robustness and thermodynamic stability tests and the average of globul size was 65.4 nm. The SNEDDS formulation containing black seed oil has good physical characteristic and stability.
The bark of cinnamon (Cinnamomum burmannii) contains cinnamaldehyde and other active substances with potent antioxidant properties. Antioxidants are effective at preventing and reducing UV-induced skin damages and skin aging. This study was intended to formulate and characterize the antioxidant peel-off facial masks containing cinnamon bark extract and the combination of polyvinyl alcohol (PVA) and hydroxypropyl methylcellulose (HPMC) as gelling agents. The ethanol extract of cinnamon bark and the developed peel-off mask were evaluated for their antioxidant activities by the α,α-diphenyl-β-picrylhydrazyl (DPPH) free radical scavenging method and for their physical characteristics. The cinnamon bark extract exhibited a very strong antioxidant activity, as evidenced by IC50= 10.04 ± 0.08 ppm. As for the formulated peel-off mask, it had excellent physical characteristics, which were identified during organoleptic observations and pH, viscosity, spreadability, and film drying time evaluations. Similar to its constituent extract, this mask produced significantly potent antioxidant effects, with IC50= 47.31 ± 1.47 ppm. For these reasons, peel-off facial masks containing cinnamon bark extract have not only excellent physical characteristics but also powerful antioxidant properties.
Abstract. COVID-19 disease caused by infection with the SARS-CoV-2 virus has caused 568,773,510 infected cases and 6,381,643 deaths worldwide (World Health Organization). One of the most effective measures to prevent the distribution of vaccination programs. Until now, many vaccines with various basics have been developed by researchers from around the world. One of the vaccines based on the Spike protein epitope peptide from the SARS-CoV-2 B.1.1.529 virus. This study used an in silico method that was carried out by searching for TLR5 receptors that play a role in increasing immune responses obtained from the Protein Data Bank website with the PDB code 3ULV and 3 peptide sequences of the Spike virus SARS-CoV-2 variant B.1.1.529, namely HVTYVPAQEKNFTTAP, SYQTQTKSHRARSVA and AGAALQIPFAMQMAYR obtained from scientific journals, peptide modeling using the PEP-FOLD3 website, preparation of structures on Biovia Discovery Studio 2021 and docking using Hpepdock. The results of the AGAALQIPFAMQMAYR peptide have the best affinity, so it can be used as one of the basic epitopes in the design of COVID-19 vaccines. Abstrak. Penyakit COVID-19 yang disebabkan oleh infeksi virus SARS-CoV-2 telah menyebabkan 568.773.510 kasus terinfeksi dan 6.381.643 kematian di seluruh dunia (World Health Organization). Salah satu langkah yang dianggap paling efektif untuk mencegah penularan yaitu vaksinasi. Hingga saat ini banyak vaksin dengan berbagai macam basis dikembangkan oleh peneliti dari berbagai dunia. Salah satu vaksin dengan basis peptida epitop protein Spike dari virus SARS-CoV-2 B.1.1.529. Penelitian ini menggunakan metode in silico yang dilakukan dengan pencarian TLR5 yang berperan dalam meningkatkan respon imun yang diperoleh dari dari website Protein Data Bank dengan kode PDB 3ULV serta 3 urutan peptida protein Spike virus SARS-CoV-2 varian B.1.1.529 yaitu HVTYVPAQEKNFTTAP, SYQTQTKSHRRARSVA dan AGAALQIPFAMQMAYR yang diperoleh dari dari jurnal ilmiah, pemodelan peptida menggunakan website PEP-FOLD3, preparasi struktur pada Biovia Discovery Studio 2021 dan dilakukan docking menggunakan Hpepdock. Hasil peptide AGAALQIPFAMQMAYR memiliki afinitas paling baik, sehingga dapat dijadikan salah satu basis epitop dalam perancangan vaksin COVID-19.
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