Isolasi dan karakterisasi enzim selulase dari rayap (Reticuluterme Fluipes).

Abstract

Enzim selulase mampu menguraikan selulosa dengan memutus ikatan f3 (1,4) glikosida rnenghasilkan selobiosa kemudian diubah lagi menjadi monomer glukosa. Enzim selulase yang dihasilkan mikroorganisme dalam rayap seeara ekonomi sangat berguna untuk berbagai industri. Telah dilakukan isolasi dan karakterisasi enzim selulase dari rayap. Hasil isolasi difraksinasi dengan (NH4)2SO4 secara bertingkat, Fl (0-10%), F2 (10-20%), F3 (20-40%), F4 (40-60%), F5 (60-80%), dan F6 (80400%). Karakterisasi untuk menentukan pH, temperatur, dan waktu inkubasi optimum serta poly inhibisi. Glukosa yang dihasilkan dianalisa dengan metode Nelson-Somogyi menggunakan spektro¬fotometer UV-Vis. Hasil penelitian menunjukkan bahwa fraksi 4 (F4 dengan tingled kejenuhan 40¬60%) mentpunyai aktivitas spesifik terbesar 102,8899 U/mg. Berdasarkan penentuan karakteristiknya enzim ini bekerja optimum pada pH 5,2 dengan aktivitas spesifik 101,7196 U/rng; temperatur 45°C dengan aktivitas spesifik 99,3539 U/mg; dan waktu inkubasi 45 menit dengan aktivitas spesifik 100,5089 U/mg terhadap substrat CMC 0,5%. Sedangkan karakteristik kinetilmya diketahui Km = 2,40574 Ing/rnI, dan Vm4„ =134,6620 U/mg dalam waktu inkubasi optimum tanpa inhibitor Ag+; Km = 2,59183 Ingim12, 6,5523 merni., dan Vmaks = 106,9633 U/mg dalam waktu indeubasi optimum dengan inhibitor Ag+. Pengerjaan dengan inhibitor Ag+ menunjukkart karakteristik inhibisi tak bersaing dan adanya gugus sulhidril pada rantai polipeptida enzim selulase. Cellulase enzyme can change cellulose yields selobiose by breaking the 3 (1,4)-glycoside bond,' then they are changed into monomer glucose again. Cellulase enzyme that microorganism yielded on termite in economic is very useful for industries. Isolation and characterization of the cellulase enzyme from termite has done. The resulted isolation fractionated by (NH4)2SO4 in stages which are Fl (0-10%), F2 (10-20A), F3 (20-40%), F4 (40-60%), F5 (60-80%), and F6 (80-100%). Characterization work to find pH, temperature, optimum incubation time and pattern of inhibition of the enzyme reaction. The glucose as resultant has analized with Nelson Somogyi methode by using UV-Vis spectrophotometer. Research yields that fraction 4 (F4 with saturation degrees of 40-60%) had a biggest specific activities of 102.8899 U/mg. Characterization of the enzyme reaction provided optimum pH of 5.2 with specific activities of 101.7196 U/mg; temperature of 45°C with specific activities of 99.3539 U/mg; and incubation time of 45 minutes with spesific activities of 100.5089 U/mg for CHIC 0.5% as substrate. Determination of enzyme kinetic characteristic gave Km = 2.40574 mg/mL and \irmas = 134.6620 U/mg at optimum incubation time without Ag+ inhibitor; Km = 2.59183 ingimL, = 6.5523 mg/mL and \.7mi„ = 106.9633 U/mg at optimum incubation time with Ag+ inhibitor. Treatment with Ag+ as inhibitor showed that it was the characteristic of noncompetitive inhibition mechanism and it indicated the presence of sulfhydril group in the cellulase enzymes polypeptide chain. vi ThisIdocument is Undip Institutional Repository Collection. The author(s) or copy“,„. that UNDIP-IR may, without changing the content, translate ,ubr{iission to any medium or format for the purpose of preservation. The author(s) or copyright owner(s) also agree that UNDIP-IR may keep more than one copy of this ,4,briaission for purposes of security, back-up and preservation. ( http://eprints.undip.ac.id)