Judul: Peningkatan Aktivitas Enzim L-Arabinosa Isomerase Dari Geobacillus Stearothermophilus Dengan Teknik Site Directed Mutagenesis
Abstrak: Enzim L-arabinosa isomerase (L-AI) merupakan enzim yang berperan dalam mediasi isomerisasi antara D-galaktosa menjadi D-tagatosa. D-tagatosa, adalah pemanis yang paling mirip dalam rasa dan sifat fisik dengan sukrosa dari seluruh pemanis, selain itu juga dikenal sebagai pengganti gula alami. D-tagatosa merupakan gula langka yang bersifat malabsorbsi atau sulit diserap oleh usus halus. Keunggulan sifat tersebut yang menjadikan D-tagatosa merupakan gula langka dapat dijadikan sebagai salah satu alternatif pengganti sukrosa. Sehingga banyak metode digunakan untuk meningkatkan hasil produksi D-tagatosa dari proses enzimatik L-AI. Produksi enzim L-AI pada umumnya memiliki berbagai kendala, seperti rendahnya aktivitas enzim dari galur murni yang telah diisolasi sebelumnya. Pendekatan rekayasa genetik merupakan alternatif yang dapat dilakukan untuk mendapatkan enzim L-AI dengan aktivitas yang lebih baik, salah satunya adalah dengan melakukan mutasi pada suatu posisi tertentu untuk memperbaiki sifat enzim. Oleh karena itu penelitian ini dilakukan dengan tujuan mengkonstruksi gen araA pada vektor pRHA dan mengekspresikan protein rekombinan L-AI di sitoplasma Escherichia coli serta meningkatkan produksi D-tagatosa dengan berbagai variasi mutasi. Gen penyandi enzim L-AI telah berhasil dikloning ke dalam vektor pRHA, dan diekspresikan di sitoplasma E. coli NiCo21 dengan ukuran protein 57 kDa. Gen tersebut telah berhasil dimutasi dengan menggunakan teknik site directed mutagenesis pada titik mutasi S393T, F280N, F280L dan mutasi ganda S393T-F280L dan S393T-F280N. Lebih lanjut lagi hasil uji aktivitas enzim L-AI dari kelima mutan dan tipe liar, empat diantaranya menunjukkan adanya peningkatan aktivitas enzim L-AI. Mutan S393T memiliki aktivitas spesifik tertinggi yakni 0.518 atau 159% dan diikuti oleh mutan F280L, F280N dan S393T-F280L yakni 24.7%, 15%, dan 14% berturut-turut, dibandingkan dengan tipe liar. Sedangkan penurunan aktivitas enzim ditunjukkan pada mutan S393T-F280N dengan penurunan aktivitas 6% di bandingkan tipe liar. Mutan S393T memiliki nilai untuk dikembangkan di dunia industri.
Keyword: Geobacillus stearothermophilus, L-arabinose isomerase, site directed mutagenesis

Judul: Production and Characterization of an Arabinose Isomerase from Gene of Geobacillus stearothermophilus Local Strain
Abstrak: Arabinose isomerase (AI) is an enzyme that catalyzes isomerization of galactose to tagatose. Besides being used as a low-calorie sweeteners, tagatose has been developed as a functional food because it provides many health benefits such as promoting of weight-loss, anti-halitosis, prebiotic, treating of obesity and reducing in symptoms associated with type 2 diabetes, hyperglycemia, anemia, and hemophilia. Thermostable AIs are potential for tagatose production. AI enzymes encoded by araA gene. The araA gene Geobacillus stearothermophilus originated from Tanjung Api, Poso, Indonesia has been successfully cloned and exspressed at previously study in E. coli BL21 (DE3) pLysS. However expression level of AI still low by SDS-PAGE analysis. The E. coli BL21 was incubated in 37°C at 150 rpm. This research was conducted to optimize the araA gene expression. Result from this research showed that the medium tofu liquid waste consisting yeast extract 0.5% (TLW+YE) increased enzyme productivity. Optimation production was obtained by 16 hours induction. The purification was carried out with three steps of freeze-thaw at -70°C, heat treatment (60°C, 30 minutes) and DEAE ion exchange chromatography (elution buffer 0-1000 mM NaCl). The purified enzyme exhibited optimum activity at 60°C and pH 7. The AI activity in the presence of CaCl2 and MnCl2 was increased to 152% and 563% respectively. Heat stability of enzymes in the presence of CaCl2 and MnCl2 was increased. Half-life (t 1/2) AI in the presence 1 mM of CaCl2 and MnCl2 was increased becomes 301 and 990 minutes respectively.
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Judul: Hamiltonian formulation for linear waves motion in two layers fluid
Abstrak: The surface wave could be considered as a wave that separates two fluids, namely water and air. Based on this assumption, it is introduced the interfacial wave, a wave between two layers of fluid with different density. The formulation of interfacial waves motion begins with deriving the base equation of irrotational ideal fluid. Furthermore, according to irrotational fluid assumption, the base equation can be stated in velocity potential. In this derivation, the fluids domain is assumed to be restricted by rigid lid boundary conditions, both at the upper and lower limit. Therefore, the interfacial waves motion can be explained in a hamiltonian formulation. In the hamiltonian formulation, total energy is defined as the sum of kinetic and potential energy. The hamiltonian system is obtained from reduction of kinetic energy by using the Dirichlet Neumann Operator. The resulted kinetic energy equation is nonlinear. Therefore, this form is linearized by first part of the Taylor expansion. This linearization gives a dispersion relation of linear wave. Based on this dispersion relation, the phase speed of the linear wave depends on the density ratio of the two layers fluid.
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