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Adaptivna laboratorijska evolucija kvasovke Hanseniaspora jakobsenii za izboljšan privzem maltoze
ID Grčar, Neža (Author), ID Čadež, Neža (Mentor) More about this mentor... This link opens in a new window

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Abstract
Fermentacijski procesi pri proizvodnji piva temeljijo na uporabi kvasovk rodu Saccharomyces, saj tovrstne starterske kulture omogočajo konstantnost in učinkovitost proizvodnje. Omejevanje mikrobne raznolikosti starterskih kultur pa hkrati zmanjšuje senzorične lastnosti in kompleksnost končnih produktov. Izboljšanje arome in povečanje raznolikosti okusov je mogoče doseči z bioaromatizacijo piva, ki vključuje uporabo ne-Saccharomyces kvasovk za tvorbo arom. Potencial za bioaromatizacijo ima tudi kvasovka Hanseniaspora jakobsenii, ki pa ima šibko sposobnost rasti na pivini. Zato je bil namen magistrskega dela izboljšanje privzema maltoze kvasovke H. jakobsenii ZIM 2603 z adaptivno laboratorijsko evolucijo (ALE), ki smo jo izvajali 42 dni v mikrotitrski plošči v gojišču z maltozo, rast pa smo spremljali z merjenjem absorbance v realnem času. Za postavitev eksperimenta ALE smo predhodno določili rastne parametre kvasovke H. jakobsenii v gojiščih z glukozo in maltozo kot različnima viroma ogljika ob prisotnosti oziroma odsotnosti kisika. Sevu H. jakobsenii smo določili rastno krivuljo preko meritev absorbance ter izračunali maksimalno specifično hitrost rasti in generacijski čas. Ugotovili smo, da je kvasovka H. jakobsenii najvišjo maksimalno specifično hitrost rasti dosegla v aerobnem okolju v bogatem gojišču YPD, najnižjo maksimalno specifično hitrost rasti pa je dosegla v gojišču YP z 20-odstotno glukozo pod anaerobnimi pogoji. Rast v gojišču z maltozo je bila zelo šibka in se tudi po ALE ni izboljšala. Rezultati so pokazali, da se privzem maltoze ni izboljšal, prav tako ni prišlo do mutacij v genu MALx3 evolviranih klonov populacij.

Language:Slovenian
Keywords:kvasovke, Hanseniaspora jakobsenii, adaptivna laboratorijska evolucija, bioaromatizacija, pivo, maltoza
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[N. Grčar]
Year:2022
PID:20.500.12556/RUL-141198 This link opens in a new window
UDC:579.22/.26:582.28.23:663.4
COBISS.SI-ID:122828547 This link opens in a new window
Publication date in RUL:25.09.2022
Views:682
Downloads:184
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Secondary language

Language:English
Title:Adaptive laboratory evolution of yeast Hanseniaspora jakobsenii for improved maltose uptake
Abstract:
Fermentation processes in beer brewing are based on the use of yeast strains from the Saccharomyces genus. Such starter cultures enable consistent and efficient production of beer but limited microbial diversity in the starter cultures reduces the sensory profile and complexity of the final products. Aroma improvement and an increase in flavor diversity can be achieved by bioaromatization of beer, which involves use of non-Saccharomyces yeasts for aroma production. The yeast Hanseniaspora jakobsenii has the potential for bioaromatization, but it grows only weakly on wort. Therefore, the aim of the master's thesis was to improve the uptake of maltose by the yeast H. jakobsenii ZIM 2603 with adaptive laboratory evolution (ALE). ALE was carried out for 42 days in a microtiter plate in a medium with maltose and growth was monitored by measuring the absorbance in real time. To set up the ALE experiment, we first determined the growth parameters of the yeast H. jakobsenii in media with glucose and maltose as different carbon sources in presence or absence of oxygen. We determined the growth curve of the strain H. jakobsenii using absorbance measurements and calculated maximum specific growth rate and generation time. Our results show that the yeast H. jakobsenii reached the highest maximum specific growth rate in an aerobic environment in the rich YPD medium, while it achieved the lowest maximum specific growth rate in the YP medium containing 20% glucose under anaerobic conditions. Growth in maltose medium was very weak and did not improve even after ALE. The results show that maltose uptake did not improve and that there were no mutations in the MALx3 gene of the evolved clones of the populations.

Keywords:yeasts, Hanseniaspora jakobsenii, adaptive laboratory evolution, bioaromatization, beer, maltose

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