Judul: Pemurnian dan Karakterisasi Enzim Keratinase dari Bacillus sp. BE-1
Abstrak: The keratinase studied in this research is an extracellular protease produced by Bacillus sp. BE-1. Feather meal substrates were prepared from native chicken feather, treated by sodium hydroxide at temperature of 1000C for 15 minutes to render crude keratinase extract. The crude keratinase was then purified by 2 steps of coloumn chromatography which are Hydrophobic Interaction Chromatography (HIC) and Gel Filtration Chromatography (GFC). Butyl Sepharose (HIC) increased keratinase activity to 67-fold. Further purification using Sephacryl S200-HR (GFC) increased keratinase activity up to the range of 116-515 fold. The highest specific activity of the keratinase was 35,9 U/mg with feather meal as substrate. This keratinase can be completely inhibited by ethylene diamine tetra acetic acid (EDTA). Additon of Mn2+ increased this keratinase activity significantly (about 51-fold) while Mg2+ increased the activity slightly (about 3-fold). Zymogram analysis resulted in that the purified keratinase consisted of two proteases with molecular weight 82 kDa and >97 kDa.
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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: Penentuan Awal Bulan Baru Penanggalan Hijriah Berdasarkan Pendekatan Ethnooceanography Dan Ethnoastronomy
Abstrak: Pergerakan matahari dan bulan terhadap rotasi bumi menjadi rujukan sistem waktu di bumi, dimana pergerakan bulan menjadi dasar sistem penanggalan Hijriah (lunar system). Pemaknaan penanggalan Hijriah melalui proyeksi posisi bulan (Ethnoastronomy) dan pergerakan pasang surut (Ethnooceanography) sebagai nilai-nilai kearifan lokal telah lama diketahui dan diaplikasikan oleh masyarakat Indonesia timur. Suku Sama menerapkannya pada konstruksi rumah dan bagan tancap, Suku Muna/Buton dan Bugis dalam manajemen pelayaran, sedangkan masyarakat di Pulau Tidore mengaplikasikannya pada penentuan awal bulan Ramadhan dan Syawal. Bentuk aplikasi mereka tersebut belum pernah dilakukan kajian ilmiah, sehingga perlu untuk dikaji melalui suatu penelitian. Penelitian ini memiliki empat tujuan utama yaitu; 1) Menentukan variasi tunggang air melalui nilai amplitudo konstituen harmonik pergerakan pasang surut dari pengelompokkan data berdasarkan sistem penanggalan Masehi dan Hijriah; 2).Membuktikan karakteristik pasang surut berdasarkan Ethnooceanography dan Ethnoastronomy dari masyarakat Tidore dan Suku Pelaut; 3) Mencari dasar secara ilmiah (oseanografi) bagian dari karakteristik pergerakan vertical pasang surut dari Suku Sama dlam menentukan tunggang air dan cara Joguru Kesultanan Tidore menentukan awal bulan baru Hijriah dan 4). Membangun solusi penentuan awal bulan Hijriah bagi masyarakat di Indonesia berdasarkan karakteristik pergerakan pasang surut. Perhitungan konstituen harmonik pasang surut, data sebaiknya tersusun berdasarkan penanggalan Hijriah. Susunan data awal yang dimulai dengan fase bulan baru (penanggalan Hijriah) menghasilkan deviasi amplitudo relatif stabil dan membentuk pola yang sama sepanjang tahun. Perhitungan tunggang pasang surut dengan Metode Suku Sama (MSS) sangat effektif, hanya menggunakan dua data pengukuran yang hasilnya sama dengan perhitungan menggunakan nilai konstituen harmonik; Tunggang air atau Likkas Silapas (LS) selain bulan Sya’ban berada di bawah nilai tunggang air rata-rata atau Mean Highest Water Level (MHWL) dan pengukuran tunggang air dengan MSS tidak dapat diterapkan untuk pengukuran pasang surut di fase bulan baru. Selisih tinggi air atau Likas boe (LB) bulan Sya’ban relatif lebih tinggi dibandingkan dengan bulan lainnya; adanya kesamaan penggunaan nilai 1/3 (33 %) dari persamaan perhitungan tunggang air dengan menggunakan MSS dan dengan menggunakan selisih rasio amplitudo; Tunggang air rata-rata atau Mean Hightest Water Level (MHWL) tidak memberi pengaruh nyata tiap bulan Hijriah (Fhit < 0.5), sedangkan tunggang air tinggi tertinggi (Hight Hightest Water Level) atau HHWL memberi pengaruh yang nyata (Fhit > 0.5); Tunggang air HHWL di Bulan Sya’ban, Dzulhijjah dan Djumadil awal lebih berpengaruh nyata pada nilai tunggang air dibandingkan bulan lainnya. Cara suku pelaut (Metode Manzillah) effektif dalam mengidentifikasi waktu dalam penanggalan Hijriah dan variasi tingi air (peak) melalui penentuan posisi bulan terhadap “Rasi bintang 7 (RB7). Metode Manzillah mempunyai dasar ilmiah iii untuk menentukan variasi tinggi pergerakan pasang surut dalam periode bulan dan tahun Hijriah, makin jauh bulan dari ekuator langit (RB7) dengan deklinasi negatif (Dec -) maka tinggi air pasang surut peak I < peak II (siklus harian) pergerakan pasang surut dan sebaliknya jika deklinasi positif (Dec +) maka peak I > peak II dan posisi bulan di sekitar ekuator langit (peak I  peak II). Penentuan awal bulan baru dengan Metode Joguru (MJ) merupakan hasil “Ijtihad” akibat bulan sabit tipis (Hilal) di awal bulan baru Ramadhan dan Syawal tidak pernah terlihat di wilayah Tidore dan sekitarnya. Ada korelasi yang kuat (R2 = 0.87 –
Keyword: Metode Suku Sama, Likkas Silapas, Likkas Boe, Metode Manzillah, Rasi Bintang 7, peak, Metode Joguru, Ijtihad’, slackwater, waktu kritis, ketampakan Hilal