研究課題/領域番号 |
21K06612
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研究種目 |
基盤研究(C)
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配分区分 | 基金 |
応募区分 | 一般 |
審査区分 |
小区分47050:環境および天然医薬資源学関連
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研究機関 | 北海道大学 |
研究代表者 |
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研究期間 (年度) |
2021-04-01 – 2024-03-31
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研究課題ステータス |
交付 (2022年度)
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配分額 *注記 |
4,160千円 (直接経費: 3,200千円、間接経費: 960千円)
2023年度: 780千円 (直接経費: 600千円、間接経費: 180千円)
2022年度: 1,560千円 (直接経費: 1,200千円、間接経費: 360千円)
2021年度: 1,820千円 (直接経費: 1,400千円、間接経費: 420千円)
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キーワード | microbiomes / aquatic animals / biosynhetic genes / Bacterial cultivation / aquatic vertebrates / cultivation / biosynthesis / 3-D cultivation |
研究開始時の研究の概要 |
The important role of gut microbiomes in the prolonged lifespan of eukaryotic hosts has recently been recognized. Our proposed research aims to identify key bacteria, metabolites/natural products, and biosynthetic genes responsible for longevity and healthy lifespan in long-lived aquatic vertebrates. We believe that this has great scientific prospect, because natural products with anti-ageing properties can be developed as pharmaceutical agents that slow ageing or treat age-related diseases, thereby contributing to human healthy lifespan.
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研究実績の概要 |
It has recently been recognized that the important contribution of beneficial microbiomes to the prolonged lifespan of their animal hosts. In our efforts to identify key bacteria, metabolites and biosynthetic genes that may contribute to the longevity of aquatic animals, we initially obtained bacterial isolates from long-lived animals, such as chitons (lifespan ~40 years) and a Japanese rockfish (lifespan >90 years). We isolated and purified metagenome from each chiton sample. Subsequent PCR-cloning and sequencing led to the detection of a biosynthetically promising bacterium in the metagenome from two of five studied chiton samples. We isolated ~430 bacterial strains from different parts of individual chitons and tested them for antibiotic resistance. It was found that ~109 bacterial strains showing resistance to certain antibiotics, which were subsequently subjected to PCR-cloning to detect strains with biosynthetic potential. These screening efforts to identify biosynthetically promising bacteria are still on going. This is in collaboration with the Lab of Prof. Joern Piel (ETH Zurich) to complement with their work on the bacterial chemistry. Using known sponge-derived compounds, we optimized the screening protocol for testing the ability of compounds to stimulate lifespan extension in the organism model Saccharomyces cerevisiae. This was based on monitoring proteasome activity of yeast lysates after being treated with tested compounds in comparison with the positive control (proteasome-stimulating compounds) and the negative control (proteasome inhibitor).
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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
Research has progressed overall more than it was originally planned. A fluorogenic substrate used for proteasome activity assay was obtained commercially in extremely small amounts, which would be insufficient and expensive for high-throughput (HT) screening of many samples. Therefore, assays based on the whole yeast cells to measure replicative lifespan (RLS) or chronological lifespan (CLS) would be applied as an alternative way for the preliminary HT screening of crude extracts; and the proteasome activity assay based on fluorogenic substrate would be used only for relatively pure compounds.
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今後の研究の推進方策 |
We are still conducting screening bacterial isolates to identify biosynthetically promising bacterial strains. This screening is not only based on their effect on lifespan extension using yeast as the organism model, but also based on antibiotic resistance combined with PCR-cloning using probes targeting biosynthetic genes. We plan to perform genome sequencing to identify promising biosynthetic genes. We will also isolate bioactive compounds with lifespan effect from promising isolates.
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