Optimasi Kitooligosakarida Cangkang Rajungan ... Jurnal Agroteknologi Vol. 16 No. 01 (2022) 85 OPTIMASI KITOOLIGOSAKARIDA CANGKANG RAJUNGAN (Portunus pelagicus) MENGGUNAKAN ENZIM KITOSANASE DARI Bacillus sp. Optimization of Chitooligosacharides of Cramb Shell (Portunus pelagicus) Using Chitosanase Enzyme from Bacillus sp. -Septi Lilik Yuliana 1) *, Dedin Finatsiyatull Rosida 1) *, Andre Yusuf Trisna Putra 1) 1) Program Studi Teknologi Pangan, Fakultas Teknik, Universitas Pembangunan Nasional “Veteran” Jawa Timur Jalan Rungkut Madya No. 1 Surabaya, 60294, Jawa Timur, Indonesia *Korespondensi Penulis: septililikyuliana04@gmail.com; dedin.tp@upnjatim.ac.id ABSTRACT Chitooligosaccharides (COS) are glycoprotein complex compounds derived from chitosan linked by 1,4 glucosidic bonds with chain lengths of 20 or less. Crab shell is a crustacean waste that contains 20-30% chitin as the basic material for making chitooligosaccharides. Chitooligosaccharide hydrolysis can be carried out enzymatically. Enzymatic hydrolysis is specific and controlled. Enzymes that can be used to hydrolyze chitosan into chitooligosaccharides are chitosanase enzymes. Chitosanase enzyme is a glycosyl hydrolase enzyme that catalyzes the hydrolysis of -1,4 glycosidic bonds of chitosan to produce low molecular weight chitooligosaccharides. One of the bacteria that produces chitosanase enzymes, namely Bacillus sp. The purpose of this study was to optimize the production of chitooligosaccharides by treating the enzyme concentration, hydrolysis time, and hydrolysis temperature. This research was conducted using a completely randomized design (CRD) with the following factors: enzyme concentration (0.5%, 1%, 1.5%, 2%, 2.5%) and hydrolysis temperature (30 o C, 40 o C, 50 o C, 60 o C, 70 o C, 80 o C) and time of hydrolysis (1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours and 8 hours). This enzyme works optimally at pH 4.5. The results showed that this chitosanase enzyme had optimum activity at a concentration of 2% (w/w) chitosan, a temperature of 50 o C, and a hydrolysis time of 5 hours. The chitooligosaccharide has a reducing sugar content of 8.49 ± 0.38 mg/mL, a viscosity of 19.08 ± 0.01 cPs, and a molecular weight of 4.51 kDa. Keywords: chitooligosaccharides, crab shells, chitosanase enzyme, hydrolysis PENDAHULUAN Kitooligosakarida (chitooligosaccha- rides/COS) adalah potongan kitin dan kitosan senyawa kompleks golongan glikoprotein yang memiliki rantai 20 atau kurang dengan ikatan 1,4 glukosamin. Kitooligosakarida merupakan kitosan yang larut dalam air. COS dihasilkan oleh depolimerisasi kitin atau kitosan menggunakan hidrolisis asam, hidrolisis oleh metode fisik, dan degradasi enzimatik (Aam et al., 2010). Rantai ikatan kitosan ß (1,4) glikosidik dapat dipotong dengan menggunakan enzim chitoosanase (Mourya et al., 2011). Enzim kitosanase merupakan salah satu enzim kitinolitik yang menghidrolisis ikatan (1,4)-β glikosidik pada kitosan atau ikatan 2- amino-2-deoksi-β-D-glukosida pada polimer kitosan yang menghasilkan DOI: https://doi.org/10.19184/j-agt.v16i01.28259