1995 Fiscal Year Final Research Report Summary
Molecular Mechanism and Engineering of Sequential Reactions in Natural Products Biosynthesis in Plants
| Project/Area Number |
06680552
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Bioorganic chemistry
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| Research Institution | CHIBA UNIVERSITY |
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Principal Investigator |
SAITO Kazuki Chiba University, Faculty of Pharmaceutical Sciences, Professor, 薬学部, 教授 (00146705)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAZAKI Mami Chiba University, Faculty of Pharmaceutical Sciences, Lecturer, 薬学部, 講師 (70222370)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Genetic engineering / Natural products / Moleculart cloning / Biosynthesis / Amino acids / Sulfur metabolism |
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| Research Abstract |
The non-protein amino acids represent one of the major groups in secondary metabolites in plants. These compounds were synthesized via sequential reactions catalyzed by specific enzymes. The cDNA encoding b-PA/DSase was isolated from an expression cDNA library of watermelon seedlings by genetic conmplementation in Cys-E.coli lacking CSase loci. The expressed protein by the clone exhibited the activity of b-PA synthesis, indicating the identity of the clone encoding b-PA/CSase. Over-expression system of b-PA/CSase in E.coli was established by using a T7 promoter-based vector. The recombinant b-PA/CSase from watermelon was able to synthesize mimosine and quisqualic acid as well as b-PA in vitro. b-PA was also formed by in vivo experiments fed with pyrazole and serine as the biosynthetic precursors. The cDNA clone of SATase was isolated from watermelon and over-expressed in E.coli. Biochemical characterization has been performed by using the purified recombinant protein.
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