diplomsko delo univerzitetnega študijskega programa
Natalija Čurič (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor)

Abstract

Delo opisuje pripravo katalitično aktivnih zamreženih encimskih skupkov iz encima α — amilaze ali na kratko CLEA. Postopek priprave zamreženih encimskih skupkov je bil razdeljen na dva ključna dela; na obarjanje topnega oziroma nativnega encima z ustreznim organskim topilom in zamreženje tako izoborjenega encima s pomočjo mrežnega povezovalca. Sinteza zamreženih encimskih skupkov je potekala pri stalni volumski koncentraciji mrežnega povezovalca (glutaraldehida, GA) 1 % (v/v) in obarjalnega reagenta (metanola) 90 % (v/v).V našem primeru smo aktivnost CLEA iz α - amilaze določili z reakcijo hidrolize škroba topnega v vodi. Optimalni reakcijski pogoji so bili doseženi pri koncentraciji proteinov iz encimskega preparata α — amilaze: γα - amilaza = 6 mg/mL in pri koncentraciji inertnega proteina albumina (EA): γEA = 2,5 mg/mL. Reakcija hidrolize škroba je potekala 3 h. Temperaturno stabilnost zamreženih encimskih skupkov smo določili po 30 minutni izpostavitvi CLEA pri različnih temperaturah. Vpliv temperature na reakcijo hidrolize škroba imobilizirane z α — amilazo smo določili v temperaturnem razponu od 45 °C do 110 °C. Stabilnost proste α — amilaze in zamreženih encimskih skupkov iz encima α — amilaze smo preučili tudi v superkritičnem ogljikovem dioksidu. Testirali smo tudi vpliv večkratne uporabe zamreženih encimskih skupkov na njihovo stabilnost.

Keywords

immobilization;amylase;cross-linked enzyme aggregates;CLEA;glutaraldehyde;

Data

Language: English
Year of publishing:
Source: Maribor
Typology: 2.11 - Undergraduate Thesis
Organization: UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher: [N. Čurič]
UDC: 547.914+577.17(043.2)
COBISS: 14147094 Link will open in a new window
Views: 4065
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Other data

Secondary language: Slovenian
Secondary title: Imobilization of enzyme [alpha] - amylase as cross-linked enzyme aggregates (CLEA)
Secondary abstract: This work describes the preparation of catalytically active enzyme aggregates of the enzyme α – amylase or briefly CLEA. The procedure to prepare cross-linked enzyme aggregates was devided into two major steps: precipitation of soluble or native enzyme with a suitable precipitant such as organic solvent and consequently cross-linking of enzyme aggregates with the cross-linker. Synthesis of cross-linked enzyme aggregates was conducted at a constant volume concentration of the cross-linker (glutaraldehyde, GA) 1 % (v/v) and precipitant (methanol) 90 % (v/v).In the present study, the activity of CLEA from α - amylase was determined by the reaction of hydrolysis of soluble starch. The optimal reaction conditions were found to be: concentration of proteins from α – amylase enzyme preparation γα - amylase = 6 mg/mL and concentration of protein albumin (EA) γEA = 2.5 mg/mL. The hydrolysis of starch was carrier out for 3 h. Temperature stability of cross-linked enzyme aggregates was determined after exposure of the immobilized enzyme at different temperatures for 30 min. Effect of temperature on the reaction of hydrolysis of starch with immobilized α - amylase was determined in the temperature range from 45 °C to 110 °C. The stability of free α – amylase and cross – linked enzyme aggregates was finally tested in supercritical carbon dioxide. We also tested the reusability of cross – linked enzyme aggregates in consecutive cycle of hydrolysis of starch.
Secondary keywords: imobilizacija;amilaza;zamreženi encimski skupki;glutaraldehid;
URN: URN:SI:UM:
Type (COBISS): Undergraduate thesis
Thesis comment: Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages: 90 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: 1011364