Biochemical characterization of PEPC from cassava: a preliminary report
We want to understand the physiological mechanisms underlying the high photosynthetic rates of cassava under drought stress and high temperatures. Histological analysis of leaf cross sections with cassava PEPC specific antiserum to establish the enzymic compartmentalization pattem is our next goal....
| Autores principales: | , , , |
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| Formato: | Conference Paper |
| Lenguaje: | Inglés |
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International Center for Tropical Agriculture
1993
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| Acceso en línea: | https://hdl.handle.net/10568/55696 |
| _version_ | 1855532148480540672 |
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| author | López F., Y. Vélez, W. El-Sharkawy, Mabrouk A. Mayer, Jorge Edgard |
| author_browse | El-Sharkawy, Mabrouk A. López F., Y. Mayer, Jorge Edgard Vélez, W. |
| author_facet | López F., Y. Vélez, W. El-Sharkawy, Mabrouk A. Mayer, Jorge Edgard |
| author_sort | López F., Y. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | We want to understand the physiological mechanisms underlying the high photosynthetic rates of cassava under drought stress and high temperatures. Histological analysis of leaf cross sections with cassava PEPC specific antiserum to establish the enzymic compartmentalization pattem is our next goal. Phosphoenolpyruvate carboxylase (PEPC) from cassava has been purified to>95 purity by liquid chromatography (fractionated ammonium sulphate precipitation, desalting by Sephadex G-25, DEAE Sepharose ion exchange, and gel filtration through Sephacryl S-300 HR). One peak of activity was eluted from DEAE Sepharose by salt gradient at 0.125 M ammonium sulphate. Gel filtration yielded two peaks of 350 and 400 kDa, respectively. Specific activity of the main peak was 5.5 units/mg protein. PEPC activities from maize, beans, and cassava leaves were compared using a spectrophotometric assay and Fast Violet detection on polyacrylamide gels. PEPC relative content and activity in cassava have intermediate values between maize and beans. A maize PEPC specific antiserum cross-reacts with cassava PEPC, indicating homologous antigenic determinants. This has also been shown at the DNA level in hybridization studies with a maize ppc probe and total, enzyme digested cassava genomic DNA. The production of a specific antiserum will enable the conduction of histological analysis of PEPC within photosynthetic tissues using immunofluorescence techniques. |
| format | Conference Paper |
| id | CGSpace55696 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 1993 |
| publishDateRange | 1993 |
| publishDateSort | 1993 |
| publisher | International Center for Tropical Agriculture |
| publisherStr | International Center for Tropical Agriculture |
| record_format | dspace |
| spelling | CGSpace556962025-10-28T10:12:08Z Biochemical characterization of PEPC from cassava: a preliminary report López F., Y. Vélez, W. El-Sharkawy, Mabrouk A. Mayer, Jorge Edgard manihot esculenta phosphoenolpyruvate carboxylase enzymic activity electrophoresis leaves fosfenolpiruvato carboxilasa actividad enzimática electroforesis hojas We want to understand the physiological mechanisms underlying the high photosynthetic rates of cassava under drought stress and high temperatures. Histological analysis of leaf cross sections with cassava PEPC specific antiserum to establish the enzymic compartmentalization pattem is our next goal. Phosphoenolpyruvate carboxylase (PEPC) from cassava has been purified to>95 purity by liquid chromatography (fractionated ammonium sulphate precipitation, desalting by Sephadex G-25, DEAE Sepharose ion exchange, and gel filtration through Sephacryl S-300 HR). One peak of activity was eluted from DEAE Sepharose by salt gradient at 0.125 M ammonium sulphate. Gel filtration yielded two peaks of 350 and 400 kDa, respectively. Specific activity of the main peak was 5.5 units/mg protein. PEPC activities from maize, beans, and cassava leaves were compared using a spectrophotometric assay and Fast Violet detection on polyacrylamide gels. PEPC relative content and activity in cassava have intermediate values between maize and beans. A maize PEPC specific antiserum cross-reacts with cassava PEPC, indicating homologous antigenic determinants. This has also been shown at the DNA level in hybridization studies with a maize ppc probe and total, enzyme digested cassava genomic DNA. The production of a specific antiserum will enable the conduction of histological analysis of PEPC within photosynthetic tissues using immunofluorescence techniques. 1993 2015-01-28T14:21:42Z 2015-01-28T14:21:42Z Conference Paper https://hdl.handle.net/10568/55696 en Open Access International Center for Tropical Agriculture López F., Yamel; Vélez, W.; El-Sharkawy, Mabrouk A.; Mayer, Jorge Edgard. 1993. Biochemical characterization of PEPC from cassava: A preliminary report. In: Roca, William M.; Thro, Ann Marie (eds.). International Scientific Meeting Cassava Biotechnology Network (1, 1992, Cartagena de Indias, Colombia). Proceedings. Centro Internacional de Agricultura Tropical (CIAT), Cali, CO. p. 340-343. (Working document no. 123) |
| spellingShingle | manihot esculenta phosphoenolpyruvate carboxylase enzymic activity electrophoresis leaves fosfenolpiruvato carboxilasa actividad enzimática electroforesis hojas López F., Y. Vélez, W. El-Sharkawy, Mabrouk A. Mayer, Jorge Edgard Biochemical characterization of PEPC from cassava: a preliminary report |
| title | Biochemical characterization of PEPC from cassava: a preliminary report |
| title_full | Biochemical characterization of PEPC from cassava: a preliminary report |
| title_fullStr | Biochemical characterization of PEPC from cassava: a preliminary report |
| title_full_unstemmed | Biochemical characterization of PEPC from cassava: a preliminary report |
| title_short | Biochemical characterization of PEPC from cassava: a preliminary report |
| title_sort | biochemical characterization of pepc from cassava a preliminary report |
| topic | manihot esculenta phosphoenolpyruvate carboxylase enzymic activity electrophoresis leaves fosfenolpiruvato carboxilasa actividad enzimática electroforesis hojas |
| url | https://hdl.handle.net/10568/55696 |
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