ISOLASI DAN IDENTIFIKASI BAKTERI PENGHASIL ENZIM ALKALIN PROTEASE BERSUMBER DARI LIMBAH PENYAMAKAN KULIT DI PT. ADI SATRIA ABADI

Fitria Dwi Anggraini, Nanung Agus F. S.Pt., M.Sc., Ph.D.; Ir. Ambar Pertiwingrum, M.Si., Ph.D.

2015 | Skripsi | S1 PETERNAKAN

Penelitian bertujuan untuk mengisolasi dan mengidentifikasi bakteri penghasil enzim protease dari limbah cair dan padat penyamakan kulit di PT Adi Satria Abadi, Yogyakarta dan mengetahui karakterisasai aktifitas enzim yang dihasilkan. Isolasi bakteri menggunakan sampel limbah cair dan padat dari limbah penyamakan kulit yang diambil dari 4 penampungan limbah yang berbeda. Isolat bakteri ditumbuhkan pada medium dengan perbedaan pH medium (7, 8, 9, 10, 11 dan 12) untuk mengukur pertumbuhan bakteri berdasarkan ukuran diameter koloni bakteri, diameter zona bening dan indeks proteolitik, sedangkan karakterisasi aktivitas enzim diuji pada variasi pH (7, 8, 9, 10, 11 dan 12) dan suhu inkubasi (40, 50 dan 60°C). Data hasil isolasi bakteri, pertumbuhan bakteri OD 600 nm, identifikasi bakteri dan pemekatan enzim 60% ammonium sulfat dilakukan analisis secara deskriptif. Data diameter koloni, diameter zona bening, indeks proteolitik dan karakterisasi aktivitas enzim dianalis dengan menggunakan rancangan acak lengkap pola searah (Completely Randomized Design), apabila terdapat perbedaan dilanjutkan dengan uji Duncans New Multiple Range Test (DMRT). Hasil isolasi bakteri sampel kedua (L2) diuji lebih lanjut karena menunjukkan adanya aktivitas proteolitik. Identifikasi isolat bakteri L2 mempunyai morfologi koloni bulat, berwarna putih, tepian rata dan elevasi cembung, morfologi sel basil warna merah, Gram negatif, tidak motil, dan katalase positif. Diameter koloni bakteri dan diameter zona bening tertinggi diperoleh pada medium dengan pH 10 dan pH 11, namun perbedaan perlakuan tidak menunjukkan indeks proteolitik yang berbeda nyata. Aktivitas enzim tertinggi pada pH 11 dengan aktivitas unit enzim 45,18±1,77 U/ml dan aktivitas spesifik enzim 43,19±1,69 U/mg dan suhu 40°C aktivitas unit enzim 54,02±1,89 U/ml dan aktivitas spesifik enzim 51,65±1,8 U/mg. Aktivitas enzim dari protease yang terpresipitasi ammonuium sulfat 60% menunjukkan hasil yang lebih tinggi (75,8 U/ml) daripada protease kasar Kata kunci : isolasi bakteri, enzim protease, limbah penyamakan kulit, unhairing

This research was aimed to isolate and identify bacteria which produced protease enzyme from liquid and solid waste tannery in PT Adi Satria Abadi, Yogyakarta and investigate the characteristics of enzyme activity. The bacterial isolation used a sample of liquid and solid waste from leather waste which taken from a different waste reservoirs. The isolat bactery was grown in a medium with differences of pH medium (7, 8, 9, 10, 11 dan 12) to measure bacterial growth based on the measurement of bacterial colony diameter, transparent zone diameter, and index proteolitc, while the characterization of enzyme activity was done on variation of (7, 8, 9, 10, 11 dan 12) ph, with the incubation temperature in (40, 50 dan 60°C). The data from bacterial isolation, bacterial growth, bacterial identification and enzyme concentration 60% ammunium sulfate done by descriptive analysis. The data of colony diameter, diameter of transparent zone, proteolitic index and characteristic of enzyme activity analyzed by using a Complete Randomized Design in a one pattern, if differences were found in the data it will be continued with Duncan Analysis New Multiple Range Test (DMRT). The second result of bacterial isolation (L2) is tested in a further step because it showed proteolitic activity. Isolat bactery identification L2 had a circular colony morphology, white coloured, flat edge, some-shaped elevation, red basil cell morphology, negative gram, none motil, and positive catalase. The highest diameter of bacterial colony and transparent zone were on medium ph 10 and ph 11, but it did not show a significant different when it got a different treatment. The highest activity of enzyme on ph 11 with activity unit enzyme 45,18±1,77 U/ml and specific enzyme activity 43,19±1,69 U/mg and temperature of 40°C activity unit enzyme 54,02±1,89 U/ml and specific enzyme activity 51,65±1,8 U/mg the activity of enzyme from protease were precipitated ammonium sulfate 60% showed a higher result of (75,8 U/ml) rather than rough protease. Keywords: isolation of bacteria, the enzyme protease, tannery waste, unhairing.

Kata Kunci : isolasi bakteri, enzim protease, limbah penyamakan kulit, unhairing