Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa
Proteinases from buckwheat (Fagopyrum esculentum moench) seeds: Purification and properties of the 47 kDa enzyme
Апстракт
Analizirane su aspartične proteinaze semena heljde. Upotrebom pepstatin A afinitetne hromatografije, iz zrelog semena izdvojene su tri forme aspartičnih proteinaza, od 47 kDa, 40 kDa i 28 kDa, dok je u ekstraktu nezrelog semena odsustvovala forma od 40 kDa. Protein od 47 kDa naknadno je razdvojen od ostalih formi kada je hromatografiji prethodila amonijum-sulfatna precipitacija. Pokazano je da tip proteolitičkog delovanja prečišćene forme enzima odgovara delovanju himozina, aspartične proteinaze animalnog porekla, čime bi se mogla objasniti njegova sposobnost da koaguliše mleko. Enzim je lokalizovan u membranskoj ćelijskoj frakciji.
Aspartic proteinases from buckwheat seeds are analyzed. Three forms of 47 kDa, 40 kDa and 28 kDa, were purified from mature buckwheat seeds, while two forms of 47 kDa and 28 kDa were detected in developing buckwheat seeds using pepstatin A affinity chromatography. A form of 47 kDa was selectively precipitated from other forms by ammonium sulfate precipitation. This enzyme resembles the chymosin-like pattern of proteolytic activity, as it was shown using BSA and k-casein as substrates, clarifying its ability for milk-clotting. The 47 kDa aspartic proteinase form is localized in the membrane fraction.
Кључне речи:
pepstatin A affinity chromatography / buckwheat / aspartic proteinaseИзвор:
Archives of Biological Sciences, 2006, 58, 3, 171-177Издавач:
- Srpsko biološko društvo, Beograd, i dr.
Финансирање / пројекти:
- Структура, функција и регулација експресије одабраних биљних гена (RS-MESTD-MPN2006-2010-143017)
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Timotijević, Gordana AU - Radović, Svetlana R. AU - Maksimović, Vesna R. PY - 2006 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/260 AB - Analizirane su aspartične proteinaze semena heljde. Upotrebom pepstatin A afinitetne hromatografije, iz zrelog semena izdvojene su tri forme aspartičnih proteinaza, od 47 kDa, 40 kDa i 28 kDa, dok je u ekstraktu nezrelog semena odsustvovala forma od 40 kDa. Protein od 47 kDa naknadno je razdvojen od ostalih formi kada je hromatografiji prethodila amonijum-sulfatna precipitacija. Pokazano je da tip proteolitičkog delovanja prečišćene forme enzima odgovara delovanju himozina, aspartične proteinaze animalnog porekla, čime bi se mogla objasniti njegova sposobnost da koaguliše mleko. Enzim je lokalizovan u membranskoj ćelijskoj frakciji. AB - Aspartic proteinases from buckwheat seeds are analyzed. Three forms of 47 kDa, 40 kDa and 28 kDa, were purified from mature buckwheat seeds, while two forms of 47 kDa and 28 kDa were detected in developing buckwheat seeds using pepstatin A affinity chromatography. A form of 47 kDa was selectively precipitated from other forms by ammonium sulfate precipitation. This enzyme resembles the chymosin-like pattern of proteolytic activity, as it was shown using BSA and k-casein as substrates, clarifying its ability for milk-clotting. The 47 kDa aspartic proteinase form is localized in the membrane fraction. PB - Srpsko biološko društvo, Beograd, i dr. T2 - Archives of Biological Sciences T1 - Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa T1 - Proteinases from buckwheat (Fagopyrum esculentum moench) seeds: Purification and properties of the 47 kDa enzyme EP - 177 IS - 3 SP - 171 VL - 58 UR - https://hdl.handle.net/21.15107/rcub_imagine_260 ER -
@article{ author = "Timotijević, Gordana and Radović, Svetlana R. and Maksimović, Vesna R.", year = "2006", abstract = "Analizirane su aspartične proteinaze semena heljde. Upotrebom pepstatin A afinitetne hromatografije, iz zrelog semena izdvojene su tri forme aspartičnih proteinaza, od 47 kDa, 40 kDa i 28 kDa, dok je u ekstraktu nezrelog semena odsustvovala forma od 40 kDa. Protein od 47 kDa naknadno je razdvojen od ostalih formi kada je hromatografiji prethodila amonijum-sulfatna precipitacija. Pokazano je da tip proteolitičkog delovanja prečišćene forme enzima odgovara delovanju himozina, aspartične proteinaze animalnog porekla, čime bi se mogla objasniti njegova sposobnost da koaguliše mleko. Enzim je lokalizovan u membranskoj ćelijskoj frakciji., Aspartic proteinases from buckwheat seeds are analyzed. Three forms of 47 kDa, 40 kDa and 28 kDa, were purified from mature buckwheat seeds, while two forms of 47 kDa and 28 kDa were detected in developing buckwheat seeds using pepstatin A affinity chromatography. A form of 47 kDa was selectively precipitated from other forms by ammonium sulfate precipitation. This enzyme resembles the chymosin-like pattern of proteolytic activity, as it was shown using BSA and k-casein as substrates, clarifying its ability for milk-clotting. The 47 kDa aspartic proteinase form is localized in the membrane fraction.", publisher = "Srpsko biološko društvo, Beograd, i dr.", journal = "Archives of Biological Sciences", title = "Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa, Proteinases from buckwheat (Fagopyrum esculentum moench) seeds: Purification and properties of the 47 kDa enzyme", pages = "177-171", number = "3", volume = "58", url = "https://hdl.handle.net/21.15107/rcub_imagine_260" }
Timotijević, G., Radović, S. R.,& Maksimović, V. R.. (2006). Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa. in Archives of Biological Sciences Srpsko biološko društvo, Beograd, i dr.., 58(3), 171-177. https://hdl.handle.net/21.15107/rcub_imagine_260
Timotijević G, Radović SR, Maksimović VR. Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa. in Archives of Biological Sciences. 2006;58(3):171-177. https://hdl.handle.net/21.15107/rcub_imagine_260 .
Timotijević, Gordana, Radović, Svetlana R., Maksimović, Vesna R., "Aspartične proteinaze semena heljde (Fagopyrum esculentum moench) - prečišćavanje i svojstva enzima 47 kDa" in Archives of Biological Sciences, 58, no. 3 (2006):171-177, https://hdl.handle.net/21.15107/rcub_imagine_260 .