Immobilization of a-Atnylase Enzyme on the (PMMA) Poly(methyl methacrylate) Surface and Its Application for Starch Hydrolysis

• Main Author: Wicaksono, Dedy
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/38574
• Institution: Institut Teknologi Bandung

n1e immobilization of the a-amylase enzyme has been carried out on activated poly (methyl methactylate) (PMMA) surface. Activation of PMMA is needed to enable the fonnation of covalent bonds beffieen enzymes and PMMA. In this study. PMMA was activated using a 112SO. 9 M solution at 60 "C for 3 hours. Based on FTIR characterization there was no significant difference in PMMA surface FTIR spectrwn before and after activation. Then, activated PMMA is immobilized with the a-amy l ase enzyme in borate buffer 0.1 M pH 9. Based on the efiZJ me activity test on activated PMMA a positive result is indicated by the presence of reducing sugar formed . Testing of enz)-'111e activity was canied out using a starch soluti on in acetate buffer 0, I M pH 9. The e\ aluation of immobilization \\as can•ied out in the fonn of enzyme-PMMA contact time. starch substrate. starch concentration.The immobilization of the a-amylase enzyme has been carried out on activated poly (methyl methactylate) (PMMA). Activation of PMMA is needed to enable the formation of covalent bonds between enzymes and PMMA.In this study. PMMA was activated using a H 2SO.9 I\1 solution at 60 °C for 3 hours. Based on FTl R characterizati on there as no significant difference in PMMA surface FTIR spectrum before and after activation. Then, activated PMMA is immobilized with the a­ amylase enzyme in the 0.1 \It borate buffer pH 9. Based on the eOZ)'me activity test on activated PMMA a positive result is indicated by the presence of reducing sugar fanned. The e\aluation of immobilization was carried out in the f01m of enzyme-PMMA contact time. starch substrate, starch concentration. The e\aluation of PMMA-enzyme contact time was canied out from 3. 6. 12. 24 and 36 hours with optimal results at 1 2 hours. Evaluation of contact time for starch substrate was canied out at 5, l 0, 15. 30. 45, 60, 90 and 120 minutes and optimal results were obtained at 45 minutes. Testing the concentration of starch susbtrat (w I v) was carried out at optimal conditions for 1 2 homs enzyme con tact time, contact time of starch substrate -l5 minutes. up to a concentration of l 0 o (w I v) not yet sho ing optimal conditions.The reducing sugar produced increases with the increase in the concentration of starch substrate. but the observation of optimal concentration is constrained by the manufacture of starch because the higher the concentration of starch the more difficult it is to dissolve. At optimal conditions for 1 2 hours of enzyme contact. contact time for starch 45 minutes and starch concentration 0.2°o (w I v) obtained a reducing sugar concentration of 1382.2 ± 28.5 ppm per 0.5 gram of acti\ated Pt>•.lMA. The optimal condition repeat test for the second and third uses gives a value of864.2 ::1: 24.7 and 751.6 ± 12 ppm per 0.5 gram activated PMMA.