KONVERSI PENISILIN MENJADI 6-APA OLEH ENZIM PENISILIN ASILASE YANG DIAMOBILKAN DENGAN K-KARAGENAN DAN KITIN

• Main Author: Sebayang, Firman
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/1116
• Institution: Institut Teknologi Bandung

<b>ABSTRACK:</br></b><br /> <p align=\"justify\">The isolation of penicillin acylase enzyme has been done from Escherichia coll. This enzyme catalyses the hydrolysis of penicillin to 6-aminopenicillanic acid (6- APA) and phenyl acetic acid. 6-APA is used as raw material to produce semisynthetic penicillin. Spesific activity of the crude enzyme extract from Escherichia coil, which were grown in Morita and Iwata medium with 0.27. phenyl acetic acid as a inducing agent, was 0.48 unit/mg protein. After purification with ammonium sulphate 20 - 60%, the spesific activity increased to 1.06 unit/mg protein. Enzyme activity was determined by the amount of 6-APA produced from benzyl penicillin as substrate, at pH 7.5 and temperature 45°C after incubation for 30 minutes. The concentration of 6-APA was determined according to Kornfeld method. Protein concentration was measured with Lowry method. <br /> Enzyme was immobilized with k-Carrageenan and Chitin as support material. The spesific activity of penicillin acylase immobilized with k-Carrageenan was 0.87 unit/mg protein at pH 7.5 and temperature 45°C with incubation for 30 minutes, and with Chitin was 0.94 unit/mg protein in the same conditions. The results of the kinetic characteristic for free enzyme, were Vmax 0.076 }.tmol 6-APA/minute, KM 0.655 mg/ml, and K1 1.012 mg/ml with inhibitor of O.5) phenyl acetic acid. For the enzyme immobilized with k-Carrageenan, Vmax were 0.066 umol 6-APA/minute and KM 0.813 mg/ml, where as with Chitin, were Vmax 0.071 umol 6-APA/minute and KM 0.765 mg/ml. The results of this research showed that immobilized enzyme was more stable compared to the free enzyme against storage time at temperature 4°C and 27°C. Immobilized enzymes can be used repeatedly. Penicillin acylase immobilized with Chitin was more stable towards repeated use, compared to enzyme immobilized with k-Carrageenan.