Produksi N-asetilglukosamin dari kitin kulit udang menggunakan enzim kitinase intraseluler Bordetella trematum = Production of n-acetylglucosamine from chitin prepared from shrimp shell using intracellular chitinase bordetella trematum

Naomi, Stella (2020) Produksi N-asetilglukosamin dari kitin kulit udang menggunakan enzim kitinase intraseluler Bordetella trematum = Production of n-acetylglucosamine from chitin prepared from shrimp shell using intracellular chitinase bordetella trematum. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Produksi N-asetil-glukosamin secara kimiawi kurang efektif dan berbahaya bagi lingkungan, sehingga dibutuhkan produksi N-asetil-glukosamin menggunakan enzim kitinase yang diperoleh secara intraseluler dari Bordetella trematum. Tujuan dari penelitian ini adalah mengetahui karakteristik enzim kitinase intraseluler Bordetella trematum dengan mengetahui pH, suhu, rasio enzim dengan kitin koloidal dan waktu fermentasi terbaik. Enzim diuji aktivitasnya pada pH 3-9 dan suhu 25-60℃. Selanjutnya, kadar N-asetil-glukosamin yang dihasilkan diukur pada rasio enzim dengan kitin koloidal sebesar 1:1, 1:3. 1:5. 1:7, 1:9 dan pada waktu fermentasi yang berbeda-beda yaitu 1 jam, 2 jam, 3 jam dan 4 jam menggunakan metode DNS. Enzim yang digunakan adalah enzim kitinase intraseluler kasar dan semi-murni dengan purifikasi parsial menggunakan ammonium sulfat 70%. Substrat yang digunakan dalam penelitian ini adalah kitin kolodial karena memiliki afinitas yang baik dengan enzim kitinase. Diperoleh bahwa enzim kitinase intraseluler kasar dari Bordetella trematum memiliki pH, suhu, rasio enzim dengan kitin koloidal dan waktu fermentasi terbaik berturut-turut pada pH 4, 45℃, 1:1 dan 2 jam. Enzim kitinase intraseluler semi-murni dari Bordetella trematum memiliki pH, suhu, rasio enzim dengan kitin koloidal dan waktu fermentasi terbaik berturut-turut pada pH 5, 50℃, 1:3 dan 3 jam. Enzim kitinase intraseluler kasar akan menghasilkan kadar N-asetil-glukosamin sebesar 49,17±0,84 ppm sedangkan enzim kitinase intraseeluler semi-murni menghasilkan kadar N-asetil-glukosamin sebesar 54,19±0,80 ppm pada kondisi fermentasi terbaik. / N-acetyl-glucosamine production with chitinase is more effective than using chemical reactions, thus chitinase from Bordetella trematum can be use for N-acetyl-glucosamine production. The purpose of this study was to report the characterization of intracellular chitinase from Bordetella trematum to optimize the production of N-acetyl-glucosamine. The enzyme activity assay will be done with varying pH (pH 3-9) and temperature (25-60℃). The maximum production of NAG will be measured with varying ratio of enzyme:colloidal chitin (1:1. 1:3, 1:5, 1:7, 1:9) and varying fermentation time (1, 2, 3, 4 hours) using DNS method. The crude and semi-crude enzyme are used in this study. Semi-crude enzyme obtained after purification of crude enzyme with ammonium sulfate 70%. Substrat that will be used in this study is colloidal chitin that has best affinity to the enzyme. The crude intracellular chitinase enzyme from Bordetella trematum has the best pH, temperature, enzyme ratio with colloidal chitin and fermentation time at pH 4, 45 ℃, 1: 1 and 2 hours and produce 49,17±0,84 ppm N-acetyl-glucosamine. Semi-crude intracellular chitinase enzyme from Bordetella trematum has the best pH, temperature, enzyme ratio with colloidal chitin and fermentation time at pH 5, 50 ℃, 1: 3 and 3 hours and produce 54,19±0,80 ppm N-acetyl-glucosamine.

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