Immobilization of penicillin acylase from Escherichia coli on commercial sepabeads EC-EP carrier
Imobilizacija penicilin-acilaze iz Escherichia coli na komercijalnom sepabeads nosaču
dc.creator | Žuža, Milena | |
dc.creator | Šiler-Marinković, Slavica | |
dc.creator | Knežević, Zorica | |
dc.date.accessioned | 2021-03-10T10:41:54Z | |
dc.date.available | 2021-03-10T10:41:54Z | |
dc.date.issued | 2007 | |
dc.identifier.issn | 1450-7188 | |
dc.identifier.uri | http://TechnoRep.tmf.bg.ac.rs/handle/123456789/1068 | |
dc.description.abstract | This paper describes the covalent immobilization of penicillin G acylase from Escherichia coli on sepabeads EC-EP, an epoxy-activated polymethacrylic carrier and kinetic properties of the immobilized enzyme. The selected enzyme belongs to a class of biocatalysts whose industrial interest is due to their versatility to mediate hydrolysis of penicillins and semi-synthetic β-lactam antibiotics synthesis reactions. About 2.7 mg of the pure enzyme was immobilized onto each gram of sepabeads with an enzyme coupling yield of 96.9%. However, it seems that the activity coupling yield is not correlated with the amount of enzyme bound and the maximum yield of 89.4% can be achieved working at low enzyme loading (0.14 mg g-1). Immobilization of the penicillin acylase resulted in slightly different pH activity profile and temperature optima, indicating that the immobilization by this method imparted structural and conformational stability of this enzyme. It appears that both free and immobilized penicillin acylase followed simple Michaelis-Menten kinetics, implying the same reaction mechanism in both systems. | en |
dc.description.abstract | U radu je ispitana kovalentna imobilizacija penicilin-acilaze iz Escherichia coli na komercijalnom polimetakrilatnom nosaču sa epoksidnim funkcionalnim grupama (Sepabeads EC-EP) i dobijeni imobilisani enzim je okarakterisan u reakciji hidrolize penicilina. Izabrani enzim je od velikog industrijskog značaja jer katalizuje reakcije hidrolize prirodnih penicilina i sinteze polu-sintetskih β-laktamskih antibiotika. Ispitan je uticaj početne koncentracije enzima na masu i aktivnost imobilisanog enzima, kao i na maseni prinos imobilizacije i prinos aktivnosti. Najveća masa imobilisanog enzima je iznosila 2,5 mg po jedinici mase nosača, što odgovara masenom prinosu od 96,9%. Međutim, prinos aktivnosti je obrnuto proporcionalan masi imobilisanog enzima tako da se maksimalni prinos od 89,4% postiže pri imobilizaciji najmanje mase enzima (0,14 mg/g nosača). Imobilizacija enzima je prouzrokovala manje promene u pH profilu aktivnosti i optimalnim vrednostima temperature biokatalizatora, što ukazuje na neznatnu stabilizaciju enzima usled imobilizacije. Kinetika reakcije hidrolize prirodnog penicilina slobodnim i imobilisanim enzimom može se opisati Mihaelis- Mentenovom jednačinom i određene su vrednosti kinetičkih konstanti za oba sistema. | sr |
dc.publisher | Faculty of Technology, Novi Sad | |
dc.rights | openAccess | |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/4.0/ | |
dc.source | Acta periodica technologica | |
dc.subject | penicillin acylase | en |
dc.subject | covalent immobilization | en |
dc.subject | sepabeads carrier | en |
dc.subject | Michaelis-Menten kinetics | en |
dc.title | Immobilization of penicillin acylase from Escherichia coli on commercial sepabeads EC-EP carrier | en |
dc.title | Imobilizacija penicilin-acilaze iz Escherichia coli na komercijalnom sepabeads nosaču | sr |
dc.type | article | |
dc.rights.license | BY-NC-ND | |
dc.citation.epage | 182 | |
dc.citation.issue | 38 | |
dc.citation.other | (38): 173-182 | |
dc.citation.spage | 173 | |
dc.identifier.doi | 10.2298/APT0738173Z | |
dc.identifier.fulltext | http://TechnoRep.tmf.bg.ac.rs/bitstream/id/8578/1450-71880738173Z.pdf | |
dc.identifier.scopus | 2-s2.0-38149032280 | |
dc.type.version | publishedVersion |