Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit
2009
Аутори
Jurick, Wayne M., IIVico, Ivana
McEvoy, James L.
Whitaker, Bruce D.
Janisiewicz, Wojciech
Conway, William S.
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Polygalacturonase (PG) was extracted and purified from decayed 'Golden Delicious' apple fruit inoculated with Penicillium solitum. Ammonium sulfate, gel filtration, and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak corresponding to PG activity. The purified PG most likely originates from the fungus because PG activity from healthy and wounded apple tissue was undetectable. Analysis of cation exchange-purified material using sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single 50-kDa band. The enzyme was active over a broad pH range (3 to 7), with optimal activity between pH 4 and 5. PG was highly active at 20 and 37 degrees C but was also detectable at 2, 50, and 75 degrees C. Divalent cations affected PG enzyme activity; Mg and Fe increased, whereas Ca and Mn reduced activity in vitro. Thin-layer chromatographic separation of hydrolysis products and data from a PG plate activity assay based on ...staining with ruthenium red showed that the enzyme exhibits both exo and endo activity. Purified PG incubated with intact apple fruit tissue in vitro caused a 30% reduction in mass after 48 h, suggesting a role in P. solitum-mediated decay of apple fruit.
Кључне речи:
maceration / postharvest decayИзвор:
Phytopathology, 2009, 99, 6, 636-641Издавач:
- Amer Phytopathological Soc, St Paul
Финансирање / пројекти:
- USDA CRISUnited States Department of Agriculture (USDA) [1275-42430-008-00D]
DOI: 10.1094/PHYTO-99-6-0636
ISSN: 0031-949X
PubMed: 19453221
WoS: 000266213100001
Scopus: 2-s2.0-67650090922
Институција/група
Poljoprivredni fakultetTY - JOUR AU - Jurick, Wayne M., II AU - Vico, Ivana AU - McEvoy, James L. AU - Whitaker, Bruce D. AU - Janisiewicz, Wojciech AU - Conway, William S. PY - 2009 UR - http://aspace.agrif.bg.ac.rs/handle/123456789/1987 AB - Polygalacturonase (PG) was extracted and purified from decayed 'Golden Delicious' apple fruit inoculated with Penicillium solitum. Ammonium sulfate, gel filtration, and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak corresponding to PG activity. The purified PG most likely originates from the fungus because PG activity from healthy and wounded apple tissue was undetectable. Analysis of cation exchange-purified material using sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single 50-kDa band. The enzyme was active over a broad pH range (3 to 7), with optimal activity between pH 4 and 5. PG was highly active at 20 and 37 degrees C but was also detectable at 2, 50, and 75 degrees C. Divalent cations affected PG enzyme activity; Mg and Fe increased, whereas Ca and Mn reduced activity in vitro. Thin-layer chromatographic separation of hydrolysis products and data from a PG plate activity assay based on staining with ruthenium red showed that the enzyme exhibits both exo and endo activity. Purified PG incubated with intact apple fruit tissue in vitro caused a 30% reduction in mass after 48 h, suggesting a role in P. solitum-mediated decay of apple fruit. PB - Amer Phytopathological Soc, St Paul T2 - Phytopathology T1 - Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit EP - 641 IS - 6 SP - 636 VL - 99 DO - 10.1094/PHYTO-99-6-0636 ER -
@article{ author = "Jurick, Wayne M., II and Vico, Ivana and McEvoy, James L. and Whitaker, Bruce D. and Janisiewicz, Wojciech and Conway, William S.", year = "2009", abstract = "Polygalacturonase (PG) was extracted and purified from decayed 'Golden Delicious' apple fruit inoculated with Penicillium solitum. Ammonium sulfate, gel filtration, and cation exchange chromatography were used to purify the enzyme. Both chromatographic methods revealed a single peak corresponding to PG activity. The purified PG most likely originates from the fungus because PG activity from healthy and wounded apple tissue was undetectable. Analysis of cation exchange-purified material using sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed a single 50-kDa band. The enzyme was active over a broad pH range (3 to 7), with optimal activity between pH 4 and 5. PG was highly active at 20 and 37 degrees C but was also detectable at 2, 50, and 75 degrees C. Divalent cations affected PG enzyme activity; Mg and Fe increased, whereas Ca and Mn reduced activity in vitro. Thin-layer chromatographic separation of hydrolysis products and data from a PG plate activity assay based on staining with ruthenium red showed that the enzyme exhibits both exo and endo activity. Purified PG incubated with intact apple fruit tissue in vitro caused a 30% reduction in mass after 48 h, suggesting a role in P. solitum-mediated decay of apple fruit.", publisher = "Amer Phytopathological Soc, St Paul", journal = "Phytopathology", title = "Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit", pages = "641-636", number = "6", volume = "99", doi = "10.1094/PHYTO-99-6-0636" }
Jurick, W. M., I., Vico, I., McEvoy, J. L., Whitaker, B. D., Janisiewicz, W.,& Conway, W. S.. (2009). Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit. in Phytopathology Amer Phytopathological Soc, St Paul., 99(6), 636-641. https://doi.org/10.1094/PHYTO-99-6-0636
Jurick WMI, Vico I, McEvoy JL, Whitaker BD, Janisiewicz W, Conway WS. Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit. in Phytopathology. 2009;99(6):636-641. doi:10.1094/PHYTO-99-6-0636 .
Jurick, Wayne M., II, Vico, Ivana, McEvoy, James L., Whitaker, Bruce D., Janisiewicz, Wojciech, Conway, William S., "Isolation, Purification, and Characterization of a Polygalacturonase Produced in Penicillium solitum-Decayed 'Golden Delicious' Apple Fruit" in Phytopathology, 99, no. 6 (2009):636-641, https://doi.org/10.1094/PHYTO-99-6-0636 . .