| Project/Area Number |
18550143
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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 |
Environmental chemistry
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| Research Institution | Waseda University |
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Principal Investigator |
ISHII Yoshitaka Waseda University, Consolidated Research Institute for Advanced Science and Medical Care, Associate professor (70339688)
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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 |
¥3,920,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Green chemistry / Enzymatic synthesis / Structure-Activity-Relationship |
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| Research Abstract |
This research purpose is the development of novel enzymatic synthesis method of 2-hydroxyphthalic acid and 3-hydroxy-4-methyl phthalic acid as a monomer for proton-conducting polymer.
Three bacteria showing decarboxylase activity of 2-hydroxyphthalic acid were isolated and designated as WU-0402, WU-0403, and WU-0404, respectively. The 2-hydroxyphthalic acid decarboxylase activity of these strains wasinvestigated. The growing cell of WU-0404 completely degraded 3-hydroxybenzoic acid as a product by the decarboxylation of 2-hydroxyphthalic acid. On the other hand, WU-0402 and WU-0403 were stoichiometrically accumulated 3-hydroxybenzoic acid from 2-hydroxyphthalic acid. Although the carboxylation activity toward 3-hydroxybenzoic acid of three strains was investigated, no activity was shown.
Based on the conserved sequences of hydroxyl benzoic acid decarboxylase, 2-hydroxyphthalic acid decarboxylase from WU-0404 was cloned. The deduced amino acid sequence showed approximately 40% similarity toward hydroxyl benzoic acid decarboxylases that were previously reported. The recombinant E. coli expressing this gene did not show both decarboxylating toward 2-hydroxyphthalic acid and carboxylating activity toward 3-hydroxybenzoic acid.
The 2-hydroxyphthalic acid decarboxylase was purified from WU-0404 by 4 steps chromatography. However, N-terminus amino acid of this decarboxylase could not be determined, because N-terminus may be blocked.
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