Izolacija encimov iz pleurotus ostreatus

magistrsko delo

Anže Peklar (Author), Mateja Primožič (Mentor), Maja Leitgeb (Co-mentor)

Abstract

Pleurotus ostreatus ali bukov ostrigar je ena izmed najbolj kultiviranih gob v komercialne namene na svetu. Vsebnost širokega spektra encimov in drugi bioučinkovin omogoča njegovo uporabo v različnih panogah. Zaradi vedno večjega pomena pri aplikacijah učinkovin iz gob v industriji in naraščajoče ekonomske zanimivosti le-teh magistrska naloga opisuje izolacijo encimov ɑ-amilaze, lakaze, celulaze, proteaze, glukoamilaze iz ekstrakta micelijev gobe P. ostreatus z dvema različnima pristopoma, gelsko kromatografijo in SDS-PAGE elektroforezo. Analiza z gelsko kromatografija je pokazala, da je izolacija encimov možna ob predpostavki, da potovanje posameznih encimov ni ovirano zaradi neprimerne poroznosti gela, vsebnosti primesi ali pa celo medsebojnega zaviranja potovanja encimov s podobnimi molskimi masami po koloni. Problem, ki se je pojavil, je bilo neizoblikovanje vrha na kromatogramu pri elucijskem volumnu posameznega preiskovanega encima. Elucijski volumni so bili pridobljeni z eksperimentalno določenimi molekulskimi masami encimov. Kontrola učinkovitosti izolacije pa je bila izvedena z določevanjem encimske aktivnosti posameznega encima v izolatu. Ekstrakt smo nato analizirali še s SDS- PAGE elektroforezo. Ugotovili smo, da je pri 12 (w/v) % separacijskem gelu možno izolirati vse preiskovane encime, katerih molekulske mase so glede na proteinski marker sovpadale z eksperimentalno določenimi, razen proteaze in celulaze. Iz teh rezultatov sklepamo, da imata obe uporabljeni tehniki izolacije encimov potencial za aplikacije v laboratorijskem in industrijskem merilu. Z optimizacijo postopka ekstrakcije naravnih materialov iz micelija gliv ter parametrov izbrane izolacijske tehnike pa je možno celo pričakovati izolacijo z večjo ločljivostjo, encimsko aktivnostjo, selektivnostjo in čistoto produkta.

Keywords

izolacija encimov;Pleurotus ostreatus;ekstrakt;gelska kromatografija;SDS-PAGE elektroforeza;ɑ-amilaza;lakaza;celulaza;proteaza;glukoamilaza;magistrske naloge;

Data

Language:	Slovenian
Year of publishing:	2021
Typology:	2.09 - Master's Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[A. Peklar]
UDC:	543.645.4(043.2)
COBISS:	93427203
Views:	135
Downloads:	31
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Isolation of enzymes from pleurotus ostreatus
Secondary abstract:	Pleurotus ostreatus is one of the most cultivated mushrooms for commercial purposes in the world. The content of a wide range of enzymes and other bioactive substances allows its use in various industries. Due to the growing importance in the applications of active ingredients from mushrooms in industry and their growing economic interest, the master's thesis describes the isolation of enzymes ɑ-amylase, laccase, cellulase, protease, glucoamylase from P. ostreatus mycelium extract with two different approaches, gel chromatography and SDS-PAGE electrophoresis. Analysis by gel chromatography showed that the isolation of enzymes was possible on the assumption that the travel of individual enzymes was not hindered by inappropriate gel porosity, impurity content, or even mutual inhibition of enzyme column travel. The problem that arose was the non-formation of a peak on the chromatogram at the elution volume of each investigated enzyme. Elution volumes were obtained by experimentally determined molecular weights. The control of the isolation efficiency, however, was by measuring the enzyme activity. The extract was then analyzed by SDS-PAGE electrophoresis. It was found that with 12% separation gel it is possible to isolate all investigated enzymes whose molecular weights with respect to the protein marker coincided with those determined experimentally, except for protease and cellulase. From these results, we conclude that both of the enzyme isolation techniques used have potential for laboratory and industrial applications. By optimizing the acquisition and extraction of natural materials and the parameters of isolation techniques, it is even possible to expect isolation with higher resolution, enzyme activity, selectivity and purity of the product.
Secondary keywords:	enzyme isolation;Pleurotus ostreatus;extract;gel chromatography;SDS-PAGE electrophoresis;ɑ-amylase;lacase;cellulase;protease;glucoamylase;
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	1 spletni vir (1 datoteka PDF (X, 53 f.))
ID:	13884054