DOI: https://doi.org/10.14710/ijfst.19.1.29-39

PENGARUH pH, SUHU DAN JENIS SUBSTRAT TERHADAP AKTIVITAS KITINASE Bacillus sp. RNT9

*Satrio Adil Pamungkas  -  Department of Fisheries, Universitas Gadjah Mada, Jl. Flora Bulaksumur, Karang Malang, Caturtunggal, Depok, Sleman Regency, Special Region of Yogyakarta 55281, Indonesia
Indun Dewi Puspita  -  Departemen Perikanan, Universitas Gadjah Mada, Jl. Flora Bulaksumur, Karang Malang, Caturtunggal, Depok, Sleman Regency, Special Region of Yogyakarta 55281, Indonesia
Ustadi Ustadi  -  Departemen Perikanan, Universitas Gadjah Mada, Jl. Flora Bulaksumur, Karang Malang, Caturtunggal, Depok, Sleman Regency, Special Region of Yogyakarta 55281, Indonesia

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Abstract

Limbah perikanan yang berasal dari udang, kepiting, dan kerang, umumnya mengandung kitin yang merupakan suatu polisakarida. Kitin memiliki struktur polimer linier yang terdiri dari monomer β-1,4-N-asetil-D-glukosamin, dan memiliki banyak manfaat dalam bentuk produk turunannya. Produk turunan kitin, seperti glukosamin dan N-Asetilglukosamin, memiliki nilai ekonomi yang tinggi sebagai bahan baku alternatif di berbagai industri seperti farmasi dan pangan. Proses degradasi kitin menjadi senyawa tersebut dapat dilakukan melalui reaksi enzimatis dengan bantuan enzim kitinase yang diproduksi oleh bakteri. Penelitian ini bertujuan untuk mengetahui pengaruh pH (6, 7 dan 8), suhu (30ºC, 35ºC dan 40ºC) dan jenis substrat (koloidal kitin, kitin serbuk dan tepung cangkang udang) terhadap aktivitas kitinase Bacillus sp. RNT9. Parameter yang diuji adalah aktivitas kitinase (U/mL) dan kadar N-Asetilglukosamin (NAG) medium (ppm). Kedua parameter ini diukur secara kuantitatif dengan metode kolorimetri. Hasil penelitian menunjukan bahwa kondisi terbaik untuk menghasilkan aktivitas kitinase pada Bacillus sp. RNT9 adalah dengan medium pH 8, suhu inkubasi 35ºC dan jenis substrat kitin koloidal. Aktivitas kitinase tertinggi yang diproduksi Bacillus sp. RNT9 berturut-turut sebesar 0,0008 U/mL pada perlakuan pH 8 pada hari ke-2 fermentasi, 0,0013 U/mL yang diperoleh pada perlakuan suhu 35ºC hari untuk ke-4 dan 0,0010 U/mL yang diperoleh pada perlakuan jenis substrat koloidal kitin untuk hari ke-2 fermentasi. Konsentrasi NAG mencapai nilai tertinggi pada optimasi pH 8 untuk hari ke-2 fermentasi sebesar 9,5968 ppm. Pada optimasi suhu 35ºC untuk hari ke-4 fermentasi sebesar 32,387 ppm serta pada perlakuan optimasi jenis substrat koloidal kitin untuk hari ke-4 fermentasi sebesar 26,031 ppm.

 

Fishery waste originating from shrimp, crabs and clams, generally contains chitin which is a polysaccharide. Chitin has a linear polymer structure consisting of β-1,4-N-acetyl-D-glucosamine monomers, and has many benefits in the form of its derivative products. Chitin derivative products, such as glucosamine and N-acetylglucosamine, have high economic value as alternative raw materials in various industries such as pharmaceuticals and food. The process of degradation of chitin into these compounds can be carried out through enzymatic reactions with the help of chitinase enzymes produced by bacteria. This study aims to determine the effect of pH (6, 7 and 8), temperature (30ºC, 35ºC and 40ºC) and type of substrate (colloidal chitin, chitin powder and shrimp shell flour) on the chitinase activity of Bacillus sp. RNT9. The parameters tested were chitinase activity (U/mL) and medium N-Acetylglucosamine (NAG) levels (ppm). Both of these parameters were measured quantitatively by the colorimetric method. The results showed that the best conditions for producing chitinase activity in Bacillus sp. RNT9 is with a medium of pH 8, incubation temperature of 35ºC and the type of substrate is colloidal chitin. The highest chitinase activity produced by Bacillus sp. RNT9 was 0,0008 U/mL, respectively, at pH 8 treatment on the 2nd day of fermentation, 0,0013 U/mL obtained at 35ºC temperature treatment for the 4th day and 0,0010 U/mL obtained at treatment type of chitin colloidal substrate for the 2nd day of fermentation. The concentration of NAG reached the highest value at the optimization of pH 8 for the 2nd day of fermentation of 9,5968 ppm. At the optimization temperature of 35ºC for the 4th day of fermentation it was 32,387 ppm and at the optimization treatment of chitin colloidal substrate for the 4th day of fermentation it was 26,031 ppm.

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Keywords: Bacillus sp. RNT9; jenis substrat; kitinase; pH; suhu

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