Optimizacija parametrov za sintezo zamreženih encimskih skupkov iz encima holesterol oksidaze

diplomska naloga

Nataša Smrečnjak (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor)

Abstract

Delo opisuje optimizacijo ključnih parametrov za sintezo katalitično aktivnih zamreženih encimskih skupkov (CLEAs) iz holesterol oksidaze (ChOx, EC 1.1.3.6). Optimizacija reakcijskih parametrov za sintezo CLEAs je obsegala iskanje optimalnega obarjalnega reagenta, optimalne koncentracije mrežnega povezovalca, optimalne koncentracije biokatalizatorja ChOx ter optimalnega koncentracijskega razmerja med holesterol oksidazo in inertnim proteinom albuminom iz kokošjih jajc (Egg albumin, EA). Sinteza zamreženih encimskih skupkov je bila razdeljena na dva ključna dela; na obarjanje topnega oziroma nativnega biokatalizatorja z ustreznim organskim topilom in na zamreženje tako izoborjenega biokatalizatorja s pomočjo mrežnega povezovalca. V našem primeru smo uspešno sintetizirali zamrežene encimske skupke iz holesterol oksidaze. Aktivnost zamreženih encimskih skupkov smo določili z reakcijo oksidacije holesterola. Optimalni reakcijski pogoji sinteze zamreženih encimskih skupkov so bili doseženi pri koncentraciji encima ChOx 2 mg/mL in koncentraciji ogrodnega proteina albumina 2 mg/mL. Ugotovili smo, da je bila aktivnost zamreženih encimskih skupkov iz biokatalizatorja holesterol oksidaze najvišja pri koncentraciji glutaraldehida 1,8 % (v/v) in acetonu kot obarjalnemu reagentu. Končna aktivnost CLEAs iz holesterol oksidaze pri optimalnih reakcijskih pogojih je bila 94 %.

Keywords

zamreženi encimski skupki;imobilizacija;holesterol oksidaza;optimizacija;reakcijski parametri;

Data

Language:	Slovenian
Year of publishing:	2011
Source:	Maribor
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[N. Smrečnjak]
UDC:	547.914+577.17(043.2)
COBISS:	14932502
Views:	2489
Downloads:	174
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	optimization of parameters for synthesis of cross-linked enzyme clusters from cholesterol oxidase
Secondary abstract:	The study describes optimization of the key parameters for the synthesis of catalytically-active cross-linked enzyme aggregates (CLEAs), made of cholesterol oxidase (ChOx, EC 1.1.3.6). The optimization of the reaction parameters consists of seeking for the best precipitant, for the optimal concentration of the cross-linker, for the optimal enzyme concentration and for the optimal concentration ratio betwen cholesterol oxidase and inert protein albumin from chicken eggs (Egg albumin, EA). The synthesis of cross-linked enzyme aggregates was divided into two main parts; the precipitation of soluble or native enzyme with a suitable organic solvent and cross-linking of enzyme aggregates with the help of a cross-linker. Cross-linked cholesterol oxidase enzyme aggregates, which activity was determined by the reaction of oxidation of cholesterol who successfully synthesized. Optimal reaction conditions for CLEAs preparation were found at ChOx enzyme concentration of 2 mg/mL and the albumin concentration of 2 mg/mL. The higest activity recovery of CLEAs was obtained at glutaraldehyde concentracion 1,8 % (v/v) and when the acetone as a precipitant of choice was used. The final activity of CLEAs of cholesterol oxidase, achieved at the optimal reaction conditions, was 94 %.
Secondary keywords:	cross-linked enzyme aggregates;immobilization;cholesterol oxidase;optimization;reaction parameters;
URN:	URN:SI:UM:
Type (COBISS):	Undergraduate thesis
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	59 f.
Keywords (UDC):	mathematics;natural sciences;naravoslovne vede;matematika;chemistry;crystallography;mineralogy;kemija;organic chemistry;organska kemija;mathematics;natural sciences;naravoslovne vede;matematika;biological sciences in general;biologija;material bases of life;biochemistry;molecular biology;biophysics;biokemija;molekularna biologija;biofizika;
ID:	1001326