Zamreženje encimov iz gobe Pleurotus ostreatus

magistrsko delo

Mihaela Taljan (Author), Maja Leitgeb (Mentor), Mateja Primožič (Co-mentor), Katja Heržič (Co-mentor)

Abstract

V magistrskem delu je opisano gojenje glive Pleurotus ostreatus (P. ostreatus) na gojiščih različne sestave. Gojišča so bila sestavljena iz mešanice pšeničnih otrobov in odpadnih materialov. Spremljali smo prirast glive, glede na različne sestave gojišča. Tako smo ugotovili, da je najugodnejše gojišče za rast glive P. ostreatus, tisto, ki je sestavljeno iz pšeničnih otrobov in slame. Po 8. dneh gojenja P. ostreatus na različnih gojiščih smo ekstrahirali encime iz glivnega micelija P. ostreatus. V pridobljenih encimskih ekstraktih smo določili koncentracijo proteinov, aktivnosti in specifične aktivnosti encimov celulaze, lakaze in α-amilaze. Ugotovili smo, da je za proizvodnjo teh encimov iz glive P. ostreatus, najugodnejši substrat sestavljen iz pšeničnih otrobov in hruške. Magistrsko delo prav tako opisuje imobilizacijo dobljenih encimov v obliki zamreženih encimskih skupkov (CLEAs) in magnetnih zamreženih encimskih skupkov (mCLEAs). Imobilizacijo v obliki m CLEAs smo izvedli na predhodno sintetizirane magnetne nanodelce prevlečene s hitozanom (H) ali dekstranom (CMD). CLEAs in mCLEAs smo določili učinkovitost imobilizacije in preostalo specifično aktivnost. Ugotovili smo, da sta encima celulaza in lakaza bila uspešno imobilizirana v obliki CMD-mCLEAs, ki smo jih pripravili iz encimskega ekstrakta, katerega gojišče je bilo sestavljeno iz pšeničnih otrobov in trave. Imobilizacija α-amilaze pa je bila najuspešnejša v primeru sinteze H-mCLEAs, ki smo jih pripravili iz istega encimskega ekstrakta. Prav tako smo izvedli študijo stabilnosti CLEAs in mCLEAs pri 4 °C. Po 2 tednih skladiščenja pri 4 °C smo določili še preostalo specifično aktivnost CLEAs in mCLEAs. Preostala specifična aktivnost imobiliziranih encimov se je po 2. tednih skladiščenja zmanjša, kar pomeni, da je stabilnost le teh upadla.

Keywords

Pleurotus ostreatus;celulaza;lakaza;alfa-amilaza;CLEAs;mCLEAs;preostala specifična aktivnost;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:	[M. Taljan]
UDC:	582.28:664.644.5(043.2)
COBISS:	66837251
Views:	468
Downloads:	80
Average score:	0 (0 votes)
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Other data

Secondary language:	English
Secondary title:	Cross-Linking of enzymes from fungus Pleurotus ostreatus
Secondary abstract:	The master's thesis describes the cultivation of the fungus Pleurotus ostreatus (P. ostreatus) on media of different composition. The media consisted of a mixture of wheat bran and waste materials. We observed the growth of the fungus, depending on the different compositions of the medium. We found out that the most favorable medium for the growth of the fungus P. ostreatus is composed from wheat bran and straw. After 8 days, enzymes were extracted from the fungus P. ostreatus. In these enzyme extracts, protein concentration, activity and specific activity of cellulase, laccase and α-amylase enzymes were determined. We found out that for the production of these enzymes, from the fungus P. ostreatus, the most favorable substrate is pear. The master's thesis also describes the crosslinking of enzyme aggregates (CLEAs) and the magnetic crosslinking of enzyme aggregates (mCLEAs) from enzyme extracts. Magnetic crosslinking was performed using dextran magnetic nanoparticles and chitosan maghemite nanoparticles. The efficiency of immobilization and the remaining specific activity were determined by cross-linked enzyme aggregates. We found out that the enzymes cellulase and lacase were successfully immobilized in the form of CMD-mCLEAs, which were prepared from an enzyme extract whose medium consisted of wheat bran and grass. Immobilization of α-amylase was most successful in the case of the synthesis of H-mCLEAs prepared from the same enzyme extract. We were also interested in the stability of CLEAs and mCLEAs if the particles were stored at 4 °C. After 2 weeks of storage at 4 °C, the remaining specific activity of CLEAs and mCLEAs was determined. From the results, we found out that the remaining specific activity of immobilized enzymes after 2 weeks of storage is lower, which means that the stability of these enzymes has decreased.
Secondary keywords:	Pleurotus ostreatus;cellulase;laccase;alpha-amylase;CLEAs;mCLEAs;remaining specific activity;
Type (COBISS):	Master's thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	XI, 68 str.
ID:	12875970