| Project/Area Number |
18380057
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied microbiology
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| Research Institution | Kyoto University |
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Principal Investigator |
KATAOKA Michihiko Kyoto University, Grad. Sch. Agriculture, Associate Professor (90252494)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Sakayu Kyoto University, Grad. Sch, Agriculture, Professor (70093250)
OGAWA Jun Kyoto University, Grad. Sch, Agriculture, Assistant Professor (70281102)
SAKURADANI Eiji Kyoto University, Grad. Sch, Agriculture, Assistant Professor (10362427)
HAGISHITA Tairo Kyoto University, Grad. Sch, Agriculture, Associate Professor (20432346)
HIBI Makoto Kyoto University, Grad. Sch, Agriculture, Assistant Professor (30432347)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥16,410,000 (Direct Cost: ¥14,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2007: ¥7,410,000 (Direct Cost: ¥5,700,000、Indirect Cost: ¥1,710,000)
Fiscal Year 2006: ¥9,000,000 (Direct Cost: ¥9,000,000)
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| Keywords | chiral compound / carbonyl reductase / bioreduction / aminoalcohol dehydrogenase / random mutation / Agrobacterium / Rhodococcus / アミノアルコール脱水素酵素 / Hansenula |
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| Research Abstract |
Chiral compounds are useful intermediates for the production of pharmaceuticals and so on. Because of high stereospecificity and environmental aspects, biocatalytic production of chiral compounds has been attracting considerable attention. On the other hand, nitrogen-containing chiral compounds are also important for the synthesis of many useful compounds, such as pharmaceuticals and agrochemicals.
(1) 3-Quinuclidinone reductase was isolated from Agrobacterium tumefaciens, and characterized in some details. The enzyme belongs to SDR superfamily. Based on amino acid sequence of the enzyme, the gene encoding 3-quinuclidinone reductase was cloned and overexpressed in E. coli transformant cells. Furthermore, E. coli coexpressing 3-quinuclidinone reductase and glucose dehydrogenase genes was constructed, and the production system for (R)-3-quinuclidinol via asymmetric reduction with E. coil transformant cells was established.
(2) Aminoalcohol dehydrogenase from Rhodococcus erythropolis is a promising enzyme for the production of d-pseudoephedrine. However, the enzyme was inhibited by d-pseudoephedrine. To improve aminoalcohol dehydrogenase, a random mutagenesis was performed by error-prone PCR. Finally, several mutant enzymes showing the activity in the presence of high concentration of d-pseudoephedrine. Aminoalcohol dehydrogenase was induced by several aminoalcohols. Through the genetic and transcriptional analysis of aminoalcohol dehydrogenase gene, a homologous orf of the GntR family transcriptional regulators was suggested to be a negative transcriptional regulator of aminoalcohol dehydrogenase gene in R. erythropolis.
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