| 研究課題/領域番号 |
21K15427
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| 研究機関 | 東京大学 |
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研究代表者 |
Jeelani Ghulam 東京大学, 大学院医学系研究科(医学部), 助教 (60468519)
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| 研究期間 (年度) |
2021-04-01 – 2024-03-31
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| キーワード | Entamoeba histolytica / Polyamine / Protozoan parasite / Metabolisms / Drug developement |
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| 研究実績の概要 |
Expression and purification of two enzyme involved in polyamine biosynthesis pathway have been completed. Gene silencing of one polyamine biosynthesis pathway enzyme (Ornithine decarboxylase) in Entamoeba histolytica have been completed and some phenotypes (growth kinetics, cell shape) have been observed. Metabolome analysis of gene silencing transformant have been completed. Optimization of protein purification using hydrophobic (Butyl-Toyopearl) and anion exchange (MonoQ) columns have been established. A review article was submitted and accepted by Trends in parasitology (https://doi.org/10.1016/j.pt.2022.02.004)
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
Optimization of enzymatic assays (spermidine synthase, spermine synthase and others) using HPLC is taking time. Silencing of one of the novel polyamine pathway enzyme (annotated as ornithine/arginine/lysine decarboxylase in genome database) failed suggesting essential enzyme for parasite survival.
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| 今後の研究の推進方策 |
Complete functional characterization of the two enzyme which we expressed and identification of the there novel enzymatic activity.
Fluxomic based metabolomic approach in order to elucidate the whole polyamine metabolic pathway.
Purification of native E. histolytica polyamine pathway enzymes.
Transcriptional silencing and metabolome analysis of the identified enzyme and their phenotypic analysis.
Complete and submit manuscript for publication.
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| 次年度使用額が生じた理由 |
Due to the coronavirus pandemic, the planned use of the budget for conference attendance was not used.
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