| Project/Area Number |
05454073
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Osaka University (1995)
Hiroshima University (1993-1994) |
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Principal Investigator |
MUROOKA Yoshikatsu Osaka University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (60029882)
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| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Mitsuo Osaka University, Graduate School of Engineering, Assistant Professor, 工学研究科, 助手 (40220347)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥4,800,000 (Direct Cost: ¥4,800,000)
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| Keywords | Cholesterol / Cholesterol oxidase / Ketosteroid dehydrogenase / Lactic acid bacteria / Host-vector system / Fermented foods / コレステロール / ステロイド分解 / 発酵食品 / 分子育種 |
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| Research Abstract |
For the purpose of degradation of cholesterol in foods materials, we have tried to bread lactic acid bacteria having a new function to degrade cholesterol. For three years, we have devoted the following research work and obtained fruitful results.
1. Many strains of lactic acid bacteria from traditional fermented foods in Japan, Indonesia, Philippines, Thailand, Nepal, Vietnam, Mongolia, and Egypt were isolated.
2. A strain of Lactobacillus plantarum isolated from a fermented fishery product, "Burong Bangus" in Philippines was selected and analyzed plasmids. By using the fragment of plasmids from L.plantarum, two useful vectors replicated in lactic acid bacteria and Escherichia coli were constructed.
3. By using the vector constructed, we obtained the best conditions of transformation of L.plantarum strain and established host-vector system in lactic acid bacteria.
4. The cholesterol oxidase gene (choA) was recombined with the vector and the strain of L.plantarum was transformed to choA^+.
5. The genes encoded cholesterol degradating enzymes were cloned and analyzed. The genes encoded ketosteroid-DELTA^I-dehydrogenase, ketosteroid-DELTA^5-isomerase and the regulator were found.
6. These cholesterol degradation enzymes are planning to express in lactic acid bacteria. The cholesterol containing food materials will be treated by these engineered lactic acid bacteria.
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