Optimasi produksi n-asetil-glukosamin dari kulit udang windu menggunakan enzim kitinase intraseluler semi murni = Optimization of n-acetyl-glucosamine production from tiger shrimp shell using semi purified intracellular chitinase enzyme from providencia stuartii

Teja, Elisa (2018) Optimasi produksi n-asetil-glukosamin dari kulit udang windu menggunakan enzim kitinase intraseluler semi murni = Optimization of n-acetyl-glucosamine production from tiger shrimp shell using semi purified intracellular chitinase enzyme from providencia stuartii. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Kitin merupakan polisakarida yang banyak terkandung dalam kulit udang dan dapat dihidrolisis menjadi produk turunannya, yaitu N-asetil-glukosamin (NAG) yang memiliki banyak manfaat, terutama untuk mengobati radang sendi. Penelitian ini bertujuan untuk mengetahui kondisi fermentasi optimum (pH, suhu, konsentrasi substrat, dan waktu fermentasi) untuk memproduksi N-asetilglukosamin menggunakan enzim kitinase semi murni intraseluler dari bakteri Providencia stuartii. Pembuatan kitin dilakukan dengan demineralisasi dan deproteinasi kulit udang windu. Penentuan pH optimum dilakukan dengan menggunakan uji aktivitas kitinase dengan variasi pH 3, 4, 5, 6, 7, 8, dan 9 dan penentuan suhu optimum dilakukan dengan variasi suhu 30°C, 40°C, 50°C, 60°C, 70°C, dan 80°C pada kondisi pH yang telah didapat pada uji sebelumnya. Konsentrasi substrat dengan variasi 0,5%, 1%, 1,5%, dan 2% serta waktu fermentasi 2 jam, 4 jam, 6 jam, dan 24 jam digunakan untuk menentukan kondisi fermentasi optimum pada penelitian tahap II. Hasil penelitian menunjukkan bahwa pH optimum enzim kitinase adalah 5 dengan aktivitas enzim sebesar 6,68 ± 0,14 U/ml dan suhu optimum enzim kitinase adalah 40°C dengan aktivitas enzim sebesar 7,12 ± 0,19 U/ml. Dalam penelitian tahap II, kondisi fermentasi terbaik terjadi pada konsentrasi substrat 1% yang diinkubasi selama 6 jam dengan kadar NAG akhir sebesar 2.084,17 ± 28,96 ppm. / Chitin is a polysaccharide that found in shrimp shell and can be derived into N-acetyl-glucosamine which has a wide range of use, especially for treating joint damage. This research was conducted to determine the optimum fermentation condition (pH, temperature, substrate concentration, and fermentation time) to produce N-acetyl-glucosamine using semi purified intracellular chitinase from Providencia stuartii. Chitin was obtained from demineralization and deproteination of tiger shrimp shell. Optimum pH was determined using chitinase activity assay by varying pH of 3, 4, 5, 6, 7, 8, and 9 and optimum temperature was determined by varying temperature of 30°C, 40°C, 50°C, 60°C, 70°C, and 80°C at the optimum pH which has been determined. Various substrate concentration (0.5%, 1%, 1.5%, and 2%) and fermentation time (2, 4, 6, and 24 hours) was used to determine the optimum fermentation condition in the second stage research. The result showed that the optimum pH of chitinase enzyme was 5 with the enzyme activity 6.68 ± 0.14 U/ml and the optimum temperature of chitinase enzyme was 40°C with the enzyme activity 7.12 ± 0.19 U/ml. The optimum fermentation condition was achieved from the fermentation at 1% substrate concentration and incubated for 6 hours with the final NAG concentration of 2,084.17 ± 28.96 ppm.

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