Karakterisasi enzim kitinase ekstraseluler dari Salmonella enterica untuk produksi N-asetilglukosamin dari kitin udang vaname. = Extracellular chitinase enzyme characterization from Salmonella enterica for the production of N-acetylglucosamine from vaname shrimp chitin

Kusuma, Laurensia Leonnie (2020) Karakterisasi enzim kitinase ekstraseluler dari Salmonella enterica untuk produksi N-asetilglukosamin dari kitin udang vaname. = Extracellular chitinase enzyme characterization from Salmonella enterica for the production of N-acetylglucosamine from vaname shrimp chitin. Bachelor thesis, Universitas Pelita Harapan.

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Abstract

Cangkang udang diketahui memiliki kandungan kitin yang tinggi, sehingga dapat dimanfaatkan dalam produksi senyawa turunannya seperti N-asetilglukosamin. Bakteri kitinolitik seperti Salmonella enterica diketahui mampu memproduksi kitinase yang dapat digunakan dalam memproduksi N-asetilgukosamin dari kitin. Penelitian dilakukan untuk (i) menentukan pH dan suhu optimum, (ii) memperoleh rasio antara enzim dan substrat terbaik, dan (iii) memperoleh lama inkubasi terbaik bagi kitinase ekstraseluler dari Salmonella enterica dalam memproduksi Nasetilglukosamin dari kitin udang vaname. Metode penelitian yang dijalankan merupakan metode eksperimental yang terdiri atas 2 tahap. Tahap I digunakan untuk menentukan pH dengan memberi perlakuan inkubasi pada pH 3, 4, 5, 6, 7, 8, dan 9 serta menentukan suhu optimum dengan perlakuan inkubasi pada 35o , 40o , 45o , 50o , dan 55oC. Tahap II dilakukan dengan menginkubasi enzim dan substrat pada perbandingan 1:1, 1:3, 1:5, 1:7, dan 1:9 (v/v) dan dilanjutkan dengan inkubasi pada perlakuan lama inkubasi 1, 2, 3, 4, dan 5 jam. Aktivitas enzim dan kadar Nasetilglukosamin akan diukur dengan reagen DNS menggunakan spektrofotometer UV-Vis pada panjang gelombang 540 nm. Hasil analisis menunjukkan bahwa kondisi optimum diperoleh oleh kitinase ekstraseluler kasar dan semi murni pada pH 5 dan 6 serta suhu inkubasi 35oC dan 40oC dengan aktivitas enzim sebesar 1,98 ± 0,12 U/mL dan 2,45 ± 0,02 U/mL secara berurutan. Rasio enzim:substrat optimum bagi kitinase ekstraseluler adalah 1:3 untuk enzim kasar dan 1:5 untuk enzim semi murni. Lama inkubasi optimum dalam memproduksi Nasetilglukosamin bagi enzim kasar dan enzim semi murni adalah 3 jam, dengan konsentrasi N-asetilglukosamin yang dihasilkan masing-masing sebesar 3876,0 ± 78,8 ppm dan 4181,6 ± 266,1 ppm. / Shrimp shells are known to have a high chitin component that can be utilized in the production of its derivative compounds such as N-acetylglucosamine. Chitinolytic bacteria such as Salmonella enterica was discovered to be able to produce chitinase which can be used in producing N-acetylglucosamine from chitin. This study was conducted to (i) determine the optimum pH and temperature, (ii) to obtain the best ratio of enzyme dan substrate needed, dan (iii) to obtain the optimum incubation time for extracellular chitinase from Salmonella enterica in producing Nacetylglucosamine from vannamei shrimp chitin. The research methodology carried out in the study is an experimental method consisting of 2 stages. the first stage is used to determine the optimum pH by giving an incubation treatment at pH 3, 4, 5, 6, 7, 8, dan 9 as well as determining the optimum temperature from 35o , 40o , 45o , 50o dan 55oC. The second stage was done by incubating the enzyme dan substrate with a ratio of 1:1, 1:3, 1:5, 1:7, dan 1:9 (v/v), dan continued with the incubation period for 1, 2, 3, 4, dan 5 hours. Enzyme activity dan N-acetylglucosamine concentrations were measured by DNS reagents using UV-Vis spectrophotometer with wavelength 540 nm. The analysis showed that the optimum conditions were obtained by crude dan semi crude extracellular chitinases at pH 5 dan 6 with incubation temperatures of 35o C dan 40o C with enzyme activity of 1,98 ± 0,12 U/mL dan 2,45 ± 0,02 U/mL, respectively. The incubation condition was optimum at 1:3 enzyme:substrate ratio dan 3 hours of incubation for crude enzyme dan at 1:5 enzyme:substrate ratio dan 3 hours of incubation for semi crude enzyme, with the highest concentration of N-acetylglucosamine produced 3876,0 ± 78,8 ppm dan 4181,6 ± 266,1 ppm, respectively.

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