Enzim tersekat adalah pemangkin penting yang digunakan terutamanya untuk meningkatkan kadar tindak balas. Salah satu cara enzim untuk disekat gerak adalah dengan ikatan kovalen. Dalam kajian ini, Candida rugosa (C. rugosa) lipase berjaya disekat gerak oleh kaedah ikatan kovalen ke manik kitosan aktif. Sekat gerak lipase pada manik kitosan telah disahkan dengan menentukan aktiviti lipase semasa hidrolisis bahan piawai. Kesan kepekatan lipase dan glutaraldehyde (GA) terhadap hasil sekatan gerak dan hasil aktiviti disiasat. Secara umum, kepekatan GA dan lipase memberikan kesan yang signifikan terhadap hasil sekat gerak dan hasil aktiviti. Memandangkan peningkatan kepekatan GA, hasil aktiviti akan menurun, sementara semakin meningkat kepekatan lipase, hasil sekat gerak berkurang. Hasil sekat gerak tertinggi adalah 77.335 %, di mana dicapai pada 6 % daripada GA dan 0.001 g / mL daripada C. rugosa lipase. Selepas itu, aktiviti tertinggi ialah 50.3613 IU, di mana diperolehi pada 2 % daripada GA dan 0.010 g / mL daripada C. rugosa lipase.
Pengoptimuman parameter proses sekat gerak dijalankan melalui Kaedah Permukaan Respon (RSM). Telah didapati bahawa keadaan optimum untuk enzim disekat gerak adalah pada 4 % kepekatan gluatarldehyde dan 0.005 g / mL daripada C. rugosa lipase yang memberikan 65.14 % hasil imobilisasi dan 32.84 IU daripada aktiviti unit. Lipase tersekat digunakan berulang kali dalam tindak balas hidrolisis. Telah didapati bahawa enzim sekat gerak lebih stabil dan mempunyai keupayaan untuk digunakan beberapa kali. Oleh itu, lipase tersekat gerak pada sokongan chitosan diaktifkan boleh dilakukan dan boleh digunakan semula sebanyak 7 kali. Walau bagaimanapun, aktiviti ini berkurangan kepada 53.67 % selepas kitaran ke-7.
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The immobilized enzyme is an important catalyst which used mainly to increase the rate of reaction. One of the methods to immobilize the enzyme is by covalent bonding. In the present research, Candida rugosa (C. rugosa) lipase was successfully immobilized by the method of covalent bonding onto activated chitosan beads. The immobilization of lipase on the chitosan beads was confirmed by determining lipase activity during hydrolysis of a standard substance. The effect of lipase and glutaraldehyde (GA) concentrations on immobilization yield and activity yield were investigated. In general, the concentration of GA and lipase gave significant effect on immobilization yield and activity yield. As the GA concentration increase, the activity yield was decreased, while as the lipase concentration increase, the immobilization yield relatively decrease. The highest immobilization yield was 77.335 %, were achieved at 6 % of GA and 0.001 g/mL of C. rugosa lipase. After that, the highest activity was 50.3613 IU, where obtained at 2 % of GA and 0.010 g/mL of C. rugosa lipase.
The optimization of immobilization process parameters was carried out via Respond Surface Method (RSM). It has been found that the most optimal condition for immobilization was at 4 % glutaraldehyde concentration and 0.005 g/mL of C. rugosa lipase which gave 65.14 % of immobilization yield and 32.84 IU of unit activity. The immobilized lipase was used repeatedly in the hydrolysis reaction. It was found that the immobilized enzyme was more stable and has the ability to be used multiple times. Therefore, immobilization of lipase onto activated chitosan support was feasible and it can be reused for 7 times. However, the activities reduce to 53.67 % after the 7th cycle.
The immobilization of candida rugosa lipase on the chitosan beads / Muliati Mohd Romli