Imobilizacija encimov na visoko porozne polimerne nosilce

magistrsko delo

Lara Berložnik (Avtor), Muzafera Paljevac (Mentor), Peter Krajnc (Komentor)

Povzetek

Namen magistrskega dela je bil razviti učinkovite postopke za sintezo poroznih polimernih materialov v različnih oblikah, kot so monoliti in zrna. Monolitne materiale smo pripravili s polimerizacijo v masi in s polimerizacijo zunanje faze emulzij z visokim deležem notranje faze, medtem ko smo zrna sintetizirali s suspenzijsko polimerizacijo. Zrna smo nato uporabili kot nosilce za imobilizacijo encimov. Najprej smo sintetizirali monolitne poli(glicidil metakrilat-ko-etilen glikol dimetakrilatne) materiale s polimerizacijo v masi, pri čemer smo uporabili različne deleže porogenega topila, ki so variirali med 0 in 200 vol. % glede na volumen monomerov. Produkt je bil krhek material, pri čemer se je z nižjimi deleži porogena tvoril material z neporozno morfologijo, z višjimi deleži pa t.i. cvetačna morfologija. S polimerizacijo zunanje faze emulzij z visokim deležem notranje faze, inducirano termično ali s fotopolimerizacijo ter uporabo zamreževala v deležih od 5 mol % do 25 mol %, smo pridobili monolitne materiale s celično morfologijo. Pore v teh materialih so bile raznolike tako po obliki kot po velikosti. Materiale smo karakterizirali z vrstičnim elektronskim mikroskopom za določitev morfologije,FTIR spektroskopijo in elementno analizo za določitev kemijske sestave ter porozimetrijo za določitev porazdelitve velikosti por in specifične površine. Sintetizirana in okarakterizirana poli(glicidil metakrilat-ko-etilen glikol dimetakrilatna) zrna smo uporabili kot nosilce za imobilizacijo encima lipaze iz Candida cylindracea. Imobilizacijo encima smo izvedli na poroznih in neporoznih zrnih, tako z uporabo distančnika in mrežnega zamreževalca kot brez njiju. Imobilizacija je bila uspešna v vseh primerih, pri čemer je bila nekoliko bolj učinkovita ob uporabi distančnika in mrežnega zamreževalca, kar smo potrdili s testno reakcijo hidrolize 4-nitrofenil butirata. Učinkovitost vezave encima smo preverjali z določanjem koncentracije proteina pred in po imobilizaciji z Bradfordovo metodo.

Ključne besede

poliHIPE material;HIP emulzija;suspenzijska polimerizacija;imobilizacija encima;Lipaza Candida Cylindracea;magistrske naloge;

Podatki

Jezik:	Slovenski jezik
Leto izida:	2024
Tipologija:	2.09 - Magistrsko delo
Organizacija:	UM FKKT - Fakulteta za kemijo in kemijsko tehnologijo
Založnik:	[L. Berložnik]
UDK:	577.15:[54.057:678.744.33](043.2)
COBISS:	221210883
Št. ogledov:	0
Št. prenosov:	17
Ocena:	0 (0 glasov)
Metapodatki:

Ostali podatki

Sekundarni jezik:	Angleški jezik
Sekundarni naslov:	Immobilization of enzymes on highly porous polymer supports
Sekundarni povzetek:	The aim of this master thesis was to develop efficient methods for the synthesis of porous polymeric materials in various forms, such as monoliths and beads. The monolithic materials were prepared by bulk polymerization and polymerization of the outer phase of high internal phase emulsions, while the beads were synthesized by suspension polymerization. The beads were then used as carriers for the immobilization of enzymes. First, we synthesized monolithic poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) materials by bulk polymerization using different amounts of porogenic solvents ranging from 150 to 450 vol% relative to the monomer volume. The product was a brittle material, with lower porogen concentrations resulting in a non-porous morphology, while higher concentrations resulted in a so-called cauliflower morphology. By polymerizing the outer phase of high inner phase emulsions, initiated thermally or by photopolymerization, and using crosslinkers at concentrations ranging from 5 to 25 mol%, we obtained monolithic materials with cellular morphology. The pores in these materials varied in both shape and size. The materials were characterized by scanning electron microscopy to determine the morphology, by FTIR spectroscopy and elemental analysis to determine the chemical composition, and by porosimetry to determine the pore size distribution and specific surface area. The synthesized and characterized poly(glycidyl methacrylate-co-ethylene glycol dimethacrylate) beads were used as carriers for the immobilization of Candida cylindracea lipase. Immobilization of the enzyme was performed on both porous and non-porous beads, with and without the use of a spacer and crosslinker. Immobilization was successful in all cases, although a slightly higher efficiency was observed when a spacer and crosslinker were used, as confirmed by the 4-nitrophenylbutyrate hydrolysis test reaction. The efficiency of enzyme binding was evaluated by determining the protein concentration before and after immobilization using the Bradford method.
Sekundarne ključne besede:	polyHIPE material;HIP emulsion;suspension polymerization;enzyme immobilization;Candida cylindracea lipase;
Vrsta dela (COBISS):	Magistrsko delo/naloga
Komentar na gradivo:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Strani:	1 spletni vir (1 datoteka PDF (XIII, 66 f.))
ID:	25054585