Biotehnološka proizvodnja lakaz in njihova uporaba pri razvoju mikrobiosenzorja

magistrsko delo študija 2. bolonjske stopnje Kemijskega inženirstva

Andreja Petrović (Author), Polona Žnidaršič Plazl (Mentor), Marjan Marinšek (Thesis defence commission member), Gabriela Kalčíková (Thesis defence commission member)

Abstract

Lakaze so zaradi sposobnosti oksidacije najrazličnejših substratov industrijsko zelo zanimivi encimi, obenem pa zaradi možnosti pretvorbe številnih fenolnih spojin igrajo pomembno vlogo pri varovanju okolja ter pri razvoju biosenzorjev. Senzorji na osnovi lakaz so pomembni za analizo kvalitete voda, kjer z njimi zaznavamo prisotnost strupenih fenolnih spojin, prav tako pa se uveljavljajo na področju analize antioksidantov v živilih. Z miniaturizacijo senzorjev zmanjšamo čas analize, stroške reagentov in količino odpadkov. V pričujočem delu sem submerzno gojila glive bele trohnobe Trametes versicolor, Trametes gibbosa in Dichomitus squalens, ki so znane proizvajalke izvenceličnih lakaz. V filtratih produkcijskih gojišč sem ovrednotila lakazno aktivnost ter njeno odvisnost od pH vrednosti. Izbrani filtrat z najvišjo vsebnostjo lakaz sem uporabila za reakcijo oksidacije v mikropretočnem sistemu, v katerega sem uspešno integrirala miniaturiziran pretočni spektrofotometrični detektor in senzor za raztopljeni kisik. Rezultati in-line meritev produkta oksidacije na izstopu iz mikroreaktorja so primerljivi z meritvami v konvencionalnem spektrofotometru, kar je dobra osnova za nadaljnji razvoj miniaturiziranega pretočnega biosenzorja na osnovi lakaz.

Keywords

glive bele trohnobe;encimi;lakaze;encimska aktivnost;submerzno gojenje;mikropretočni sistemi;biosenzorji;magistrska dela;

Data

Language:	Slovenian
Year of publishing:	2021
Typology:	2.09 - Master's Thesis
Organization:	UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher:	[A. Petrović]
UDC:	604.4:577.15(043.2)
COBISS:	53985795
Views:	328
Downloads:	64
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Bioctehnological laccases production and their use in a microbiosensor development
Secondary abstract:	Laccases are industrially relevant enzymes because of their ability to oxidize a wide variety of substrates. At the same time, they play an important role in environmental protection and biosensor development due to their ability to transform many phenolic compounds. Laccase-based sensors are important for water analysis, where they detect the presence of toxic phenolic compounds, and are also used in the food industry to detect antioxidants. By miniaturizing the sensors, we reduce analysis time, reagent costs and waste. In the present work, the submerged cultivation of the white rot fungi Trametes versicolor, Trametes gibbosa and Dichomitus squalens, which are known producers of extracellular laccase, was studied. Laccase activity in the filtrate of the production medium and its dependence on pH were evaluated. The filtrate with the highest laccase activity was used for the oxidation reaction in the microfluidic system, in which a miniaturized flow spectrophotometric detector and an oxygen microsensor were successfully integrated. The results of in-line measurements of the oxidation product at the outlet of the microreactor were comparable to the measurements in a conventional spectrophotometer, which provides a good basis for further development of a miniaturized laccase-based flow-through biosensor.
Secondary keywords:	laccase;enzyme activity;submerged cultivation;microfluidic system;biosensor;
Type (COBISS):	Master's thesis/paper
Study programme:	1000376
Thesis comment:	Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo
Pages:	XVI, 45 str.
ID:	12612162