| Project/Area Number |
14560283
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物資源科学
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| Research Institution | Osaka Prefecture University |
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Principal Investigator |
MIYATAKE Kazutaka Osaka Prefecture University, Department of Applied Biological Chemistry, Professor, 農学生命科学研究科, 教授 (70094513)
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| Co-Investigator(Kenkyū-buntansha) |
UEDA Mitsuhiro Osaka Prefecture University, Department of Applied Biological Chemistry, Lecturer, 農学生命科学研究科, 講師 (50254438)
NAKAZAWA Masami Osaka Prefecture University, Department of Applied Biological Chemistry, Assistant, 農学生命科学研究科, 助手 (90343417)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2004: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2002: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Metabolic Engineering / Citrate / Citrate transporter / Polyglutamic Acid / Bacillus natto / Gene Transportation / トランスポーター / 遺伝子操作 / バチルス / 大腸菌 / 機能発現 / 大量生産 |
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| Research Abstract |
We investigated the citric acid transporter genes in Bacillus and cloned some of the genes. We expressed these genes in E.coli and measured the citrate transport activities. These transport activities were raised in special cultivation conditions, especially the aeration and magnesium concentration. Furthermore, we investigated the incorporation of citric acid through transporters in genes introduced into bacteria, Bacillus natto. We found the polyglutamic acid production was increased by the introduction of the citric acid transporter gene. We also found that the cultivation conditions, especially the oxygen density, the metal ions, especially magnesium, and the extract of yeast influenced the citric acid incorporation in the transformed bacteria.
A 4-5 times productivity was obtained in comparison with the control production conditions.
Introduction of a yraO gene that is thought as a citric acid transporter gene from the genomes of the bacillus, also increased the growth and incorporation of citric acid in medium.
Incorporation of heavy metals in the bacterial cells were accelerated with the co-presence of citric acid, this suggests that this method is useful for heavy metal purification in an polluted environment, the so called bioremediation.
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