Susanto, Tius Enggarsari (2014) Isolasi dan Purifikasi Enzim Kitinase dari Isolat Bakteri Tanah Limbah Industri Petis Udang dengan Metode Afinitas Kitin. Sarjana thesis, Universitas Brawijaya.
Abstract
Indonesia merupakan negara kepulauan dengan sebagian besar wilayahnya berupa perairan. Industri berbasis perairan di Indonesia telah banyak berkembang salah satunya adalah pengolahan udang. Menurut kementrian kelautan dan perikanan (2013), total produksi udang pada tahun 2012 mencapai 415.703 ton. Limbah pengolahan udang yang terdiri dari kulit, ekor dan kepala udang mencapai sekitar 30% dari satu ekor udang. Limbah udang memiliki potensi yang besar untuk diolah menjadi produk kitin. Kitin merupakan polisakarida yang tersusun atas β-1,4-N-asetil-D-glukosamin (GlcNac). Pada umumnya kitin berikatan dengan protein dan mineral. Kitin memiliki struktur yang rigid dan tidak dapat larut dalam air sehingga kitin menjadi sumber pencemaran senyawa organik. Berdasarkan permasalahan di atas, dilakukan isolasi dan skrining mikroba kitinolitik dari tanah limbah industri petis udang di Sidoarjo, Jawa Timur. Mikroba kitinolitik yang diperoleh mampu menghasilkan enzim kitinase yang mampu mendegradasi kitin sehingga dapat mengurangi tingkat pencemaran senyawa organik. Selanjutnya enzim kitinase yang diperoleh dilakukan karakterisasi. Penelitian ini menggunakan metode deskriptif. Hasil penelitian dipaparkan berdasarkan tahapan penelitian yang dilakukan. Hasil penelitian menunjukkan bahwa terdapat tiga isolat mikroba yang ditunjukkan dengan adanya zona bening di sekitar koloni mikroba. Indeks kitinolitik tertinggi ditunjukkan oleh isolat TP02 sebesar 2,22. Aktivitas spesifik ekstrak enzim kasar sebesar 0,008 Unit/mg dan enzim murni sebesar 0,052 Unit/mg. Kadar protein ekstrak enzim kasar 3,636 mg/ml dan enzim murni sebesar 1,127 mg/ml. Tingkat kemurnian enzim kitinase murni mencapai 6,548 kali. Berat molekul enzim kitinase dari isolat TP02 dengan menggunakan SDSPAGE sekitar 68,257 kDa. Enzim kitinase isolat TP02 memiliki pH optimum 7, suhu optimum 35°C, kestabilan pH 5-7 dan kestabilan suhu 30-45°C. Enzim kitinase isolat TP02 spesifik terhadap substrat kitin. Produk hidrolisis substrat dideteksi dengan metode kromatografi lapis tipis. Hasil hidrolisis berupa N-asetil- D-glukosamin (GlcNAc) dan GlcNAcn.
English Abstract
Indonesia is an archipelago with the most of its territory is in the form of sea. Industrial based of sea in Indonesia has been evolved, one of them is the shrimps processing. According to the ministry of maritime affairs and fisheries (2013), total of shrimps production in 2012 reached 415.703 tonnes. Shrimp processing wastes, consisting of skin, tail and head of the shrimp reach about 30% of a shrimp. Shrimp wastes has great potential to be processed into product of chitin. Chitin is the polysaccharide composed by β-1,4-N-acetyl-D-glucosamine (GlcNAc). In general, chitin is binding with protein and mineral. Chitin has a rigid structure and it cannot soluble in water therefore that chitin becomes the source contamination of organic compounds. Based on that problem, it needs to do the isolation and screening of chitinolytic microbe from shrimp paste waste soil industry in Sidoarjo , East Java. Chitinolytic microbes that is obtained able to produce chitinase enzyme which able degrade chitin in order to reduce the contamination level of organic compounds. Furthermore, chitinase enzyme that already obtained will be characterized. This research uses the descriptive method. The results is presented based on the step of the research. The results showed that there are three microbial isolates that indicated by a clear zone around the colonies of microbes. The highest chitinolytic index is indicated by isolate TP02 that is 2.22. The specific activity of crude enzyme extracts is 0,008 units / mg and the pure enzyme is 0.052 units / mg. The protein content of crude enzyme extract is 3.636 mg / ml and the pure enzyme is 1.127 mg / ml. The purification of the chitinase was increased to 6,548 fold. The molecular weight of chitinase enzymes of isolate TP02 determined by SDSPAGE is about 68.257 kDa. Chitinase enzymes of isolate TP02 has the optimum pH is 7, optimum temperature is 35ºC, pH stability is pH 5-7 and the termal stability is 30-45º. Chitinase enzymes of isolate TP02 specific to substrate chitin. The hydrolysis product of substrate is detected by thin layer chromatography method. The hydrolysis products are N-acetyl-D-glucosamin (GlcNAc) and GlcNAcn.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/FTP/2014/302/051404852 |
| Uncontrolled Keywords: | kitinase, limbah udang, mikroba kitinolitik, tanah limbah industri petis udang,- chitinase, chitinolytic microbe, shrimp paste waste soil industry, shrimp waste |
| Subjects: | 300 Social sciences > 338 Production > 338.1 Agriculture |
| Divisions: | Fakultas Teknologi Pertanian > Teknologi Hasil Pertanian |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 21 Aug 2014 14:40 |
| Last Modified: | 02 Dec 2021 04:21 |
| URI: | http://repository.ub.ac.id/id/eprint/149701 |
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