Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes
The promastigote stage of Leishmania resides in the sand fly gut, enriched with sugar molecules. Recently we reported that Leishmania donovani possesses a sucrose uptake system and a stable pool of intracellular sucrose metabolizing enzyme. In the present study, we purified the intracellular sucrase...
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Wiley
2016-01-01
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| Series: | Biochemistry Research International |
| Online Access: | http://dx.doi.org/10.1155/2016/7108261 |
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| author | Arpita Singh Debjani Mandal |
| author_facet | Arpita Singh Debjani Mandal |
| author_sort | Arpita Singh |
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| description | The promastigote stage of Leishmania resides in the sand fly gut, enriched with sugar molecules. Recently we reported that Leishmania donovani possesses a sucrose uptake system and a stable pool of intracellular sucrose metabolizing enzyme. In the present study, we purified the intracellular sucrase nearly to its homogeneity and compared it with the purified extracellular sucrase. The estimated size of intracellular sucrase is ~112 kDa by gel filtration chromatography, native PAGE, and substrate staining. However, in SDS-PAGE, the protein is resolved at ~56 kDa, indicating the possibility of a homodimer in its native state. The kinetics of purified intracellular sucrase shows its higher substrate affinity with a Km of 1.61 mM than the extracellular form having a Km of 4.4 mM. The highly specific activity of intracellular sucrase towards sucrose is optimal at pH 6.0 and at 30°C. In this report the purification and characterization of intracellular sucrase provide evidence that sucrase enzyme exists at least in two different forms in Leishmania donovani promastigotes. This intracellular sucrase may support further intracellular utilization of transported sucrose. |
| format | Article |
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| institution | Kabale University |
| issn | 2090-2247 2090-2255 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
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| series | Biochemistry Research International |
| spelling | doaj-art-69ee738be3544d9dbdebc91f784bf98a2025-02-03T05:50:17ZengWileyBiochemistry Research International2090-22472090-22552016-01-01201610.1155/2016/71082617108261Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani PromastigotesArpita Singh0Debjani Mandal1Division of Infectious Diseases and Immunology, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata, West Bengal 700032, IndiaDivision of Infectious Diseases and Immunology, Indian Institute of Chemical Biology, 4 Raja S.C. Mullick Road, Jadavpur, Kolkata, West Bengal 700032, IndiaThe promastigote stage of Leishmania resides in the sand fly gut, enriched with sugar molecules. Recently we reported that Leishmania donovani possesses a sucrose uptake system and a stable pool of intracellular sucrose metabolizing enzyme. In the present study, we purified the intracellular sucrase nearly to its homogeneity and compared it with the purified extracellular sucrase. The estimated size of intracellular sucrase is ~112 kDa by gel filtration chromatography, native PAGE, and substrate staining. However, in SDS-PAGE, the protein is resolved at ~56 kDa, indicating the possibility of a homodimer in its native state. The kinetics of purified intracellular sucrase shows its higher substrate affinity with a Km of 1.61 mM than the extracellular form having a Km of 4.4 mM. The highly specific activity of intracellular sucrase towards sucrose is optimal at pH 6.0 and at 30°C. In this report the purification and characterization of intracellular sucrase provide evidence that sucrase enzyme exists at least in two different forms in Leishmania donovani promastigotes. This intracellular sucrase may support further intracellular utilization of transported sucrose.http://dx.doi.org/10.1155/2016/7108261 |
| spellingShingle | Arpita Singh Debjani Mandal Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes Biochemistry Research International |
| title | Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes |
| title_full | Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes |
| title_fullStr | Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes |
| title_full_unstemmed | Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes |
| title_short | Purification and Characterization of a Novel Intracellular Sucrase Enzyme of Leishmania donovani Promastigotes |
| title_sort | purification and characterization of a novel intracellular sucrase enzyme of leishmania donovani promastigotes |
| url | http://dx.doi.org/10.1155/2016/7108261 |
| work_keys_str_mv | AT arpitasingh purificationandcharacterizationofanovelintracellularsucraseenzymeofleishmaniadonovanipromastigotes AT debjanimandal purificationandcharacterizationofanovelintracellularsucraseenzymeofleishmaniadonovanipromastigotes |