Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract
Izolovanje i delimična karakterizacija kisele fosfataze ekstrakta polena Artemisia vulgaris
2002
Аутори
Ćirković-Veličković, TanjaGavrović-Jankulović, Marija
Bukilica, MN
Mandić, Ljuba M.
Petrović, SZ
Jankov, Ratko M.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
An acid phosphatase from an extract of mugwort (Artemisia vulgaris) pollen was purified by a factor of 48 by a combination of ion exchange and gel-chromatography. The molecular weights of the enzyme were 76 kDa and 73 kDa, determined by gel filtration on a Sephadex G-100 sf column and by SDS PAGE (under reducing and non-reducing conditions), respectively. In analytical isoelectrofocusing, the enzyme appears as two very close bands, pI at about 4.2. The optimum pH for the enzyme is 5.4. The apparent K-m for p-nitrophenyl phosphate was estimated to be 0.16 mM. The purified enzyme has broad specificity, and hydrolyses p-nitrophenyl phosphate and alpha-naphthyl phosphate. Pyrophosphate and O-phospho-L-tyrosine were estimated to be the best substrates for this enzyme as potential in vim substrates. The enzyme is inhibited competitively by phosphate (K-i 1.25 mM), molybdate (K-i = 0.055 mM) and pyrophosphate (K-i = 6.7 mM) and non-competitively by fluoride (K-i = 9.8 mM). Metal ions such as ...Hg2+, Cu2+ and Zn2+ express an inhibitory effect on the enzyme, while the enzyme is slightly activated by non-ionic detergents, Tween 20 and Triton X-100. There is no change in the enzyme activity in the presence of tartrate, citrate, EDTA, 1,10-phenanthroline and sulfhydryl-group modifiers such as p-chloromercuribenzoate and N-ethylmaleimide.
Kisela fosfataza ekstrakta polena visokog korova (Artemisia vulgaris) je prečišćena 48 puta kombinacijom jonoizmenjivačke i gel-hromatografije. Molekulska težina enzima je 76 kDa i 73 kDa, određena gel-filtracijom na matriksu Sephadex G-100 sf i SDS PAG elektroforezom (pri redukujućim i neredukujućim uslovima), respektivno. Pri izoelektrofokusiranju, enzim se sastoji iz dve vrlo bliske trake pI vrednosti oko 4,2. Optimalno pH za aktivnost enzima je 5,4. PrividnoKmza hidrolizu p-nitrofenil-fosfata je procenjeno da je 0,16 mM. Prečišćeni enzim ima široku specifičnost hidrolizuje p-nitrofenil-fosfat i α-naftil-fosfat. Pirofosfat i O-fosfo-L-tirozin su procenjeni kao najbolji od potencijalnih in vivo supstrata ovog enzima. Enzim je inhibiran kompetitivno fosfatom (Ki=1,25 mM), molibdatom (Ki=0,055 mM) i pirofosfatom (Ki=6,7 mM) a nekompetitivno fluoridom (Ki= 9,8mM). Joni metala, kao što su Hg2+, Cu2+ i Zn2+ iskazuju inhibitorni efekat na enzim, dok je efekat ne-jonskih detergenata, kao št...o su Tween 20 i Triton X-100 blago aktivirajuć i. Nema promene u aktivnosti enzima u prisustvu tartarata, citrata, EDTA 1,10-fenantrolina i modifikatora sulfhidrilnih grupa kao što su p-hloromerkuribenzoat i N-etilmaleimid.
Кључне речи:
acid phosphatase / Artemisia vulgaris / Artemisia vulgaris / compositae / Compositae / mugwort / mugwort / pollen / pollen / purification / purificationИзвор:
Journal of the Serbian Chemical Society, 2002, 67, 8-9, 567-572Издавач:
- Serbian Chemical Soc, Belgrade
DOI: 10.2298/JSC0209567C
ISSN: 0352-5139
WoS: 000178039500003
Scopus: 2-s2.0-0036378021
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Ćirković-Veličković, Tanja AU - Gavrović-Jankulović, Marija AU - Bukilica, MN AU - Mandić, Ljuba M. AU - Petrović, SZ AU - Jankov, Ratko M. PY - 2002 UR - https://cherry.chem.bg.ac.rs/handle/123456789/508 AB - An acid phosphatase from an extract of mugwort (Artemisia vulgaris) pollen was purified by a factor of 48 by a combination of ion exchange and gel-chromatography. The molecular weights of the enzyme were 76 kDa and 73 kDa, determined by gel filtration on a Sephadex G-100 sf column and by SDS PAGE (under reducing and non-reducing conditions), respectively. In analytical isoelectrofocusing, the enzyme appears as two very close bands, pI at about 4.2. The optimum pH for the enzyme is 5.4. The apparent K-m for p-nitrophenyl phosphate was estimated to be 0.16 mM. The purified enzyme has broad specificity, and hydrolyses p-nitrophenyl phosphate and alpha-naphthyl phosphate. Pyrophosphate and O-phospho-L-tyrosine were estimated to be the best substrates for this enzyme as potential in vim substrates. The enzyme is inhibited competitively by phosphate (K-i 1.25 mM), molybdate (K-i = 0.055 mM) and pyrophosphate (K-i = 6.7 mM) and non-competitively by fluoride (K-i = 9.8 mM). Metal ions such as Hg2+, Cu2+ and Zn2+ express an inhibitory effect on the enzyme, while the enzyme is slightly activated by non-ionic detergents, Tween 20 and Triton X-100. There is no change in the enzyme activity in the presence of tartrate, citrate, EDTA, 1,10-phenanthroline and sulfhydryl-group modifiers such as p-chloromercuribenzoate and N-ethylmaleimide. AB - Kisela fosfataza ekstrakta polena visokog korova (Artemisia vulgaris) je prečišćena 48 puta kombinacijom jonoizmenjivačke i gel-hromatografije. Molekulska težina enzima je 76 kDa i 73 kDa, određena gel-filtracijom na matriksu Sephadex G-100 sf i SDS PAG elektroforezom (pri redukujućim i neredukujućim uslovima), respektivno. Pri izoelektrofokusiranju, enzim se sastoji iz dve vrlo bliske trake pI vrednosti oko 4,2. Optimalno pH za aktivnost enzima je 5,4. PrividnoKmza hidrolizu p-nitrofenil-fosfata je procenjeno da je 0,16 mM. Prečišćeni enzim ima široku specifičnost hidrolizuje p-nitrofenil-fosfat i α-naftil-fosfat. Pirofosfat i O-fosfo-L-tirozin su procenjeni kao najbolji od potencijalnih in vivo supstrata ovog enzima. Enzim je inhibiran kompetitivno fosfatom (Ki=1,25 mM), molibdatom (Ki=0,055 mM) i pirofosfatom (Ki=6,7 mM) a nekompetitivno fluoridom (Ki= 9,8mM). Joni metala, kao što su Hg2+, Cu2+ i Zn2+ iskazuju inhibitorni efekat na enzim, dok je efekat ne-jonskih detergenata, kao što su Tween 20 i Triton X-100 blago aktivirajuć i. Nema promene u aktivnosti enzima u prisustvu tartarata, citrata, EDTA 1,10-fenantrolina i modifikatora sulfhidrilnih grupa kao što su p-hloromerkuribenzoat i N-etilmaleimid. PB - Serbian Chemical Soc, Belgrade T2 - Journal of the Serbian Chemical Society T1 - Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract T1 - Izolovanje i delimična karakterizacija kisele fosfataze ekstrakta polena Artemisia vulgaris VL - 67 IS - 8-9 SP - 567 EP - 572 DO - 10.2298/JSC0209567C ER -
@article{ author = "Ćirković-Veličković, Tanja and Gavrović-Jankulović, Marija and Bukilica, MN and Mandić, Ljuba M. and Petrović, SZ and Jankov, Ratko M.", year = "2002", abstract = "An acid phosphatase from an extract of mugwort (Artemisia vulgaris) pollen was purified by a factor of 48 by a combination of ion exchange and gel-chromatography. The molecular weights of the enzyme were 76 kDa and 73 kDa, determined by gel filtration on a Sephadex G-100 sf column and by SDS PAGE (under reducing and non-reducing conditions), respectively. In analytical isoelectrofocusing, the enzyme appears as two very close bands, pI at about 4.2. The optimum pH for the enzyme is 5.4. The apparent K-m for p-nitrophenyl phosphate was estimated to be 0.16 mM. The purified enzyme has broad specificity, and hydrolyses p-nitrophenyl phosphate and alpha-naphthyl phosphate. Pyrophosphate and O-phospho-L-tyrosine were estimated to be the best substrates for this enzyme as potential in vim substrates. The enzyme is inhibited competitively by phosphate (K-i 1.25 mM), molybdate (K-i = 0.055 mM) and pyrophosphate (K-i = 6.7 mM) and non-competitively by fluoride (K-i = 9.8 mM). Metal ions such as Hg2+, Cu2+ and Zn2+ express an inhibitory effect on the enzyme, while the enzyme is slightly activated by non-ionic detergents, Tween 20 and Triton X-100. There is no change in the enzyme activity in the presence of tartrate, citrate, EDTA, 1,10-phenanthroline and sulfhydryl-group modifiers such as p-chloromercuribenzoate and N-ethylmaleimide., Kisela fosfataza ekstrakta polena visokog korova (Artemisia vulgaris) je prečišćena 48 puta kombinacijom jonoizmenjivačke i gel-hromatografije. Molekulska težina enzima je 76 kDa i 73 kDa, određena gel-filtracijom na matriksu Sephadex G-100 sf i SDS PAG elektroforezom (pri redukujućim i neredukujućim uslovima), respektivno. Pri izoelektrofokusiranju, enzim se sastoji iz dve vrlo bliske trake pI vrednosti oko 4,2. Optimalno pH za aktivnost enzima je 5,4. PrividnoKmza hidrolizu p-nitrofenil-fosfata je procenjeno da je 0,16 mM. Prečišćeni enzim ima široku specifičnost hidrolizuje p-nitrofenil-fosfat i α-naftil-fosfat. Pirofosfat i O-fosfo-L-tirozin su procenjeni kao najbolji od potencijalnih in vivo supstrata ovog enzima. Enzim je inhibiran kompetitivno fosfatom (Ki=1,25 mM), molibdatom (Ki=0,055 mM) i pirofosfatom (Ki=6,7 mM) a nekompetitivno fluoridom (Ki= 9,8mM). Joni metala, kao što su Hg2+, Cu2+ i Zn2+ iskazuju inhibitorni efekat na enzim, dok je efekat ne-jonskih detergenata, kao što su Tween 20 i Triton X-100 blago aktivirajuć i. Nema promene u aktivnosti enzima u prisustvu tartarata, citrata, EDTA 1,10-fenantrolina i modifikatora sulfhidrilnih grupa kao što su p-hloromerkuribenzoat i N-etilmaleimid.", publisher = "Serbian Chemical Soc, Belgrade", journal = "Journal of the Serbian Chemical Society", title = "Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract, Izolovanje i delimična karakterizacija kisele fosfataze ekstrakta polena Artemisia vulgaris", volume = "67", number = "8-9", pages = "567-572", doi = "10.2298/JSC0209567C" }
Ćirković-Veličković, T., Gavrović-Jankulović, M., Bukilica, M., Mandić, L. M., Petrović, S.,& Jankov, R. M.. (2002). Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract. in Journal of the Serbian Chemical Society Serbian Chemical Soc, Belgrade., 67(8-9), 567-572. https://doi.org/10.2298/JSC0209567C
Ćirković-Veličković T, Gavrović-Jankulović M, Bukilica M, Mandić LM, Petrović S, Jankov RM. Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract. in Journal of the Serbian Chemical Society. 2002;67(8-9):567-572. doi:10.2298/JSC0209567C .
Ćirković-Veličković, Tanja, Gavrović-Jankulović, Marija, Bukilica, MN, Mandić, Ljuba M., Petrović, SZ, Jankov, Ratko M., "Isolation and partial characterization of an acid phosphatase from Artemisia vulgaris pollen extract" in Journal of the Serbian Chemical Society, 67, no. 8-9 (2002):567-572, https://doi.org/10.2298/JSC0209567C . .