Judul: Study on keratinase and disulphide reductase activity from Bacillus sp. MTS in keratin degradation
Abstrak: Keratin is insoluble and very stable protein. The disulphide bonds, hydrogen bonds and hydrophobic interaction within the polypeptide gives mechanical strength and also resistance to some protease enzyme. The main objective of this research was to study degradation of native chicken feather by keratinolytic enzymes from Bacillus sp., that is the role of keratinase and disulphide reductase, and this was conducted in four steps. The first step was conducted to select Bacillus sp. which produced extracellular keratinase and disulphide reductase. The isolates used were B. licheniformis MB-2 and Bacillus sp. MTS. The first isolate was a thermophillic bacterium screened from hot springs water of Mount Tompaso-Manado and the latter was mesophillic bacterium from sulphuric soil of Mount Tangkuban Perahu- Bandung. The result showed that Bacillus sp.MTS was more effective than B.licheniformis MB-2 for chicken feather degradation. A minimum medium which contained mineral salts was the best medium for Bacillus sp.MTS to secrete disulphide reductase, while the best medium for keratinase production was minimum medium with addition of 0.02% tripton and 0.02% yeast extract. Bacillus sp.MTS was found to secrete intracellular and extracellular keratinase and disulphide reductase and it was selected for further study. The second step was conducted to characterize the crude keratinase and disulphide reductase of Bacillus sp.MTS . The optimum temperature for keratinase and disulphide reductase were 55oC and 28oC respectively. Optimum pHs for keratinase were 8.0, 10.0 and 11.0, and optimum pH for disulphide reductase was 9.0. SDS-PAGE analysis of the culture-free cell showed 17 – 60 kD protein bands. Zymography analysis of ammonium sulphate fraction (50% w/v) showed that Bacillus sp.MTS secreted multy fraction keratinases of 17, 25, 32, 53, 96 and > 97 kD. The 25 kD keratinase was the most stable enzyme at pH 8.0-11.0 and temperature 55-900 The third step was to purify keratinase and disulphide reductase. Purification with ammonium sulphate 50% (w/v), dialysis, Butyl sepharose FF chromatography dan Sephacryl S-200HR chromatography could increase keratinase and disulphide purities, and gave six active keratinase with specific activity of 10 to 37 U/mg and three active disulphide reductase with specific activity of 30 to 64 U/mg. SDS-PAGE analysis of the highest activities of keratinase fractions showed molecular weights of 53, 32 and 17 kD. The highest activities of disulphide reductase fractions showed of 29 and 17 kD molecular weight. The purified keratinase and disulphide reductase had optimum activity at alkaline pHs (8.0-12.0). C while the 32 and 53 kD fractions were found as unstable. In the fourth step the role of disulphide reductase, reducing agent (DTT and BMT) and urea in hydrolysis of native chicken feather and sheep wool and NaOH prehydrolized chicken feather were studied. The activity of purified keratinase was increased by the presence of purified disulphide reductase.
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Judul: Optimasi produksi enzim keratinase dari bakteri termofilik L-23 asal Sulawesi Utara
Abstrak: Bulu ayam merupakan limbah terbesar industri unggas. Sebagai limbah, bulu ayam sebenarnya dapat dimanfaatkan karena masih tingginya kandungan protein. Namun pemanfaatan bulu ayam menjadi terbatas karena sebagian besar bulu ayam tersusun atas keratin yang secara struktural sulit diuraikan. Salah satu cara untuk memecah bulu ayam adalah secara enzimatik, yaitu dengan menggunakan mikroba penghasil enzim keratinase. Keratinase adalah enzim yang dapat memecah substrat keratin. Salah satu isolat koleksi laboratorium Mikrobiologi dan Biokimia PAU adalah isolat L-23 asal Sulawesi Utara, yang merupakan bakteri temofilik penghasil enzim keratinase. Suhu optimum keratinase L-23 berkisar antara 60-65°C dan 75-80°C. Keratinase kasar isolat L-23 memiliki pH optimum pada 7 dan 12, sedang enzim murninya 8 dan 12. Tujuan dari penelitian ini adalah mengoptimasi produksi enzim keratinase dari bakteri termofilik, L23, yang diisolasi dari Sulawesi Utara, mencoba memproduksi enzim keratinase tersebut secara berulang dan melakukan analisis zimogram untuk melihat bentuk enzim yang dihasilkan. Untuk mengoptimasi produksi keratinase dari isolat L-23, bakteri termofilik asal Sulawesi Utara, perlu dilakukan penentuan umur inokulum optimum, media optimum dan variasi komponennya, konsentrasi substrat, pengujian pengaruh senyawa kofaktor dan keberadaan oksigen, dan cara produksi optimum. Dari penentuan umur inokulum didapatkan bahwa umur inokulum optimum adalah 10 jam. Media optimum untuk isolat L-23 adalah seperti media yang digunakan untuk degradasi keratin oleh Fervidobacterium pennavorans dengan sedikit manipulasi yaitu jumlah tripton dijadikan 0% dan ditambah glukosa 0.01%. Adapun komposisi media tersebut (per liter) adalah: 3.2 g KH2PO4. 2.0 g NaH2PO4, 1.5 g (NH4)2SO4, 3.0 g NaCI, 0.1 g CaCl₂. 2 H₂O, 0.3 g MgSO4. 7H₂O, 6 mg FeCl3, 1 g NaHCO3, 10 ml larutan mineral, dan 10 ml larutan vitamin. Komposisi larutan mineral adalah (per liter) : 1.0 g MnCl2. 4 H₂O. 1.0 g CoCl2. 6 H₂O, 0.5 g NiCl2. 6 H₂O, 0.5 g ZnCl2, 0.5 g CuSO4, 0.2 g H3BO3, 0.1 g Na2M0O4.2 H2O. Sedangkan larutan vitamin terdiri dari 1 tablet vitamin Becombion yang dilarutkan dalam 1 liter akuades. ...
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Judul: (Kasus Desa Situ Ilir, Kecamatan Cibungbulang, Kabupaten Bogor, Propinsi J Awa Barat )
Abstrak: Pengesahan UU No.22 tahun 1999 sebagai tanda pemberlakuan otonomi daerah memberikan harapan baru bagi pemerintahan Indonesia. Namun pada sisi yang lain ada tanda tanya besar apakah gerak pembaruan dalam konsep otonomi daerah yang digulirkan ini akan benar-benar selamat sampai tujuan dan sesuai dengan kehendak rakyat selama ini ataukah sebaliknya.
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