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Določanje encimske aktivnosti lipaz v ekstraktih semen in kalčkov pri različnih vrednostih pH in temperature
ID Junkar, Tajda (Author), ID Poklar Ulrih, Nataša (Mentor) More about this mentor... This link opens in a new window, ID Skrt, Mihaela (Comentor)

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Abstract
Namen magistrskega dela je bil raziskati vpliv različnih temperaturnih in pH pogojev na aktivnost lipaz iz izbranih ekstraktov semen in kalčkov, ki so na voljo na slovenskem trgu. Zastavili smo si tri delovne hipoteze, in sicer, da je aktivnost lipaz odvisna od temperature, da bo najvišja pri nevtralnem pH, višja v ekstraktih semen kot v kalčkih ter se bo razlikovala glede na izvorni rastlinski vir. Analizirali smo semena in kalčke črne soje, gorčice, sončnice, pšenice, sladke koruze, oljne redkve ter semena fižola nežike,sezama in lanene kalčke. V vzorcih smo določili vsebnost vlage oz., delež suhe snovi, koncentracijo proteinov, katero smo določali spektrofotometrično ter izvedli gelsko NaDS elektroforezo. Ugotovili smo, da vsi vzorci vsebujejo proteine v velikosti od 10 do 72 kDa. Delež suhe snovi je večji v vzorcih semen, kot kalčkov, delež proteinov na suho snov pa je v večini vzorcev večji v kalčkih, kot v istovrstnih semenih. Največ proteinov smo določili v ekstraktu semen črne soje (31,7 mg/mL), najmanj pa v ekstraktu gorčičnih kalčkov (0,65 mg/mL). Aktivnost lipaz smo preračunali iz spektrofotometričnih meritev absorbance pri 400 nm, pri pH 7,2 in temperaturnih območjih 15 °C, 25 °C, 37 °C in 45 °C ter pri različnih pH območjih, in sicer pH 4,0, 6,5, 7,2 in 9,0 ter temperaturi 37 °C. Najvišjo aktivnost lipaz smo določili v ekstraktu gorčičnih in pšeničnih kalčkov (0,26 U/mL) pri 45 °C. Najnižjo aktivnost smo določili pri 15 °C v ekstraktu kalčkov sladke koruze (0,0019 U/mL). Aktivnost lipaz se z višjo temperaturo in višjim pH povečuje,iz česar lahko zaključimo, da imajo lipaze iz analiziranih vzorcev višjo aktivnost v bazičnem okolju, medtem ko je kislo okolje (pH 4,0) za lipaze iz izbranih vzorcev bilo najmanj ugodno, saj je bila aktivnost pod mejo detekcije.

Language:Slovenian
Keywords:encimi, lipaze, semena, kalčki, rastlinski ekstrakti, encimska aktivnost
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Publisher:[T. Junkar]
Year:2024
PID:20.500.12556/RUL-164464 This link opens in a new window
UDC:577.15:547.9
COBISS.SI-ID:213101827 This link opens in a new window
Publication date in RUL:26.10.2024
Views:69
Downloads:25
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Secondary language

Language:English
Title:Determination of lipase enzyme activity in seed and germ extracts at different pH and temperature values
Abstract:
The purpose of this master's thesis was to investigate the influence of different temperature and pH conditions on the activity of lipases from selected extracts of seeds and sprouts available on the Slovenian market. We formulated three working hypotheses: first, that lipase activity depends on temperature and is highest at neutral pH; second, that it is higher in seed extracts than in sprouts; and third, that it varies depending on the plant source. We examined seeds and sprouts of black soybeans, mustard, sunflower, wheat, sweet corn, oil radish, nežika beans and sesame seeds and flaxseed sprouts. In the samples, the moisture content or dry matter content was determined, the protein concentration was measured spectrophotometrically, and SDS-PAGE gel electrophoresis was performed. We found that all samples contained proteins ranging from 10 to 72 kDa. The dry matter content was higher in seed samples than in sprouts, and the protein content on dry matter was mostly higher in sprout than in corresponding seeds. The highest protein content was determined in the black soybean seed extract (31,7 mg/mL), and the lowest in mustard sprout extract (0,65 mg/mL). Lipase activity was calculated from spectrophotometric measurements of absorbance at 400 nm at pH 7,2 and temperatures of 15 °C, 25 °C, 37 °C, and 45 °C, as well as at different pH values, pH 4,0, 6,5, 7,2, and 9,0, at temperature of 37 °C. The highest lipase activity was observed in mustard and wheat sprout extracts (0,26 U/mL) at 45 °C. The lowest activity was observed at 15 °C in sweet corn sprout extract (0,0019 U/mL). Lipase activity increased with higher temperature and pH. From these results, we concluded that lipases from the analyzed samples exhibited higher activity in a basic environment (pH 9,0), while acidic conditions (pH 4,0) were the least favorable and resulted in activity below the detection limit.

Keywords:enzymes, lipases, seeds, sprouts, plant extracts, enzyme activity

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