Sinteza zamreženih encimskih skupkov iz encima transglutaminaze

Lesičar, Špela

| | SLO | ENG | Cookies and privacy

Bigger font | Smaller font

First page > Show document

Show document

Title:	Sinteza zamreženih encimskih skupkov iz encima transglutaminaze
Authors:	ID Lesičar, Špela (Author) ID Leitgeb, Maja (Mentor) More about this mentor... ID Vasić, Katja (Comentor)
Files:	UN_Lesicar_Spela_2017.pdf (1,88 MB) MD5: 114337D8E7D66C0924FA1E89E7D9C42D PID: 20.500.12556/dkum/87913e25-685c-4003-98a8-313435f518f8
Language:	Slovenian
Work type:	Bachelor thesis/paper
Typology:	2.11 - Undergraduate Thesis
Organization:	FKKT - Faculty of Chemistry and Chemical Engineering
Abstract:	Namen diplomske naloge je bila uspešna sinteza zamreženih encimskih skupkov (CLEAs) iz encima transglutaminaze. Pri tem smo želeli doseči čim višjo učinkovitost imobilizacije in preostalo aktivnost encima transglutaminaze. Sinteza je zajemala postopek obarjanja encima v izbranem obarjalnem reagentu in zamreženja encima z mrežnim povezovalcem glutaraldehidom (GA). Da bi dosegli čim boljše rezultate smo morali izbrati ustrezen obarjalni reagent ter njegovo količino. Optimirali smo parametre pri zamreženju, kot so koncentracija dodanega mrežnega povezovalca, koncentracija dodanega natrijevega cianoborohidrida (NaBH3CN), koncentracija ogrodnih proteinov govejega serumskega albumina (BSA) in jajčnega albumina (EA) ter koncentracija transglutaminaze. Iz rezultatov smo ugotovili, da je 2-propanol najbolj ustrezen obarjalni reagent za sintezo CLEAs iz transglutaminaze. Optimalni volumski procent obarjalnega reagenta je bil 92,4 % (v/v), kar znaša 1215 μl. Za optimalne rezultate smo uporabili raztopino encima s koncentracijo 150 mg/ml s 100 mg/ml EA. Encimske molekule smo zamrežili z 2 % (v/v) GA ob 3 urnem mešanju. Za boljšo stabilnost encima smo dodali 200 μl 0,1 M NaBH3CN. Ugotovili smo, da lahko tako imobilizirano transglutaminazo kot katalizator reakcije uporabimo trikrat z aktivnostjo encima nad 100 % preden se deaktivira.
Keywords:	transglutaminaza, zamreženi encimski skupki, imobilizacija, mrežni povezovalec, glutaraldehid
Place of publishing:	Maribor
Publisher:	[Š. Lesičar]
Year of publishing:	2017
PID:	20.500.12556/DKUM-67603
UDC:	543.645.4:544.478.3(043.2)
COBISS.SI-ID:	21033494
NUK URN:	URN:SI:UM:DK:CCNTOXBG
Publication date in DKUM:	18.09.2017
Views:	1626
Downloads:	124
Metadata:
Categories:	KTFMB - FKKT
:	Copy citation

Average score:	(0 votes)
Your score:	Voting is allowed only for logged in users.
Share:

Hover the mouse pointer over a document title to show the abstract or click on the title to get all document metadata.

Licences

License:	CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International

Link:	http://creativecommons.org/licenses/by-nc-nd/4.0/
Description:	The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.
Licensing start date:	24.08.2017

Secondary language

Language:	English
Title:	Immobilization of transglutaminase as crosslinked enzyme aggregates (CLEAs)
Abstract:	The purpose of this diploma thesis was a successful synthesis of cross-linked enzyme aggregates (CLEAs) from enzyme transglutaminase. The aim was to achieve the maximum possible efficiency of immobilization and the highest remaining activity of transglutaminase. Synthesis of CLEAs consisted of precipitation of the enzyme in chosen precipitant and cross-linking of the enzyme with cross-linking reagent glutaraldehyde (GA). The best precipitant and its optimal volume for synthesis of CLEAs was selected in order to achieve the highest results and we optimized various parameters in process of crosslinking such as concentration of GA and sodium cyanoborohydride (NaBH3CN), concentracion of stabilization proteins bovine serum albumine (BSA) and egg albumin (EA) and concentration of enzyme transglutaminase. The results of diploma thesis shows that the optimal precipitant for synthesis of CLEAs from transglutaminase is 2-propanol with volume percent of 92,4 % (v/v), so the volume of percipitant is 1215 μl. 150 mg/ml of enzyme transglutaminase with 100 mg/ml of EA was used for 3 hours long process of cross-linking with 2% (v/v) GA. We added 200 μl NaBH3CN for stabilization of the enzyme. We also realized that we can reuse the immobilized transglutaminase three times before its activity falls under 100% and the enzyme deactivates.
Keywords:	transglutaminase, cross-linked enzyme aggregates, immobilization, cross-linking agent, glutaraldehyde

Comments

Leave comment

You must log in to leave a comment.

Comments (0)

0 - 0 / 0

There are no comments!

Back