| Project/Area Number |
08556016
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Bioproduction chemistry/Bioorganic chemistry
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| Research Institution | Tohoku University |
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Principal Investigator |
OHRUI Hiroshi (1997) Tohoku University, Faculty of Agriculture, Professor, 農学部, 教授 (20100050)
目黒 煕 (1996) 東北大学, 農学部, 教授 (30005590)
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| Co-Investigator(Kenkyū-buntansha) |
AKASAKA Kazuaki Tohoku University, Faculty of Agriculture, Assistant, 農学部, 助手 (10201881)
大類 洋 東北大学, 農学部, 助教授 (20100050)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | (S) -MNB-carboxylic acid / (S) -TBMB-carboxylic acid / D,L-analysis / sequential analysis / on-line HPLC-exciton CD analysis / 蛍光分析 / アミノ酸 / エナンチオマー / キラル分析 / HPLC / ベンゾジオキソ-ルカルボン酸 |
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| Research Abstract |
We have focused on the development of a highly sensitive method for the sequential determination of D,L-amino acid from the C-terminus of peptides. We have developed a method which involves the selective hydrolysis of C-terminal psptide bond as the key step. The method is as follows, (1) ethyl esterification of the C-terminal carboxylic acid, (2) NaBH_4 reduction of the ester, (3) selective hydrolysis of the terminal amide bond under acidic conditions via N*O acyl migration, (4) separation of the resulting amino alcohol from peptide and mono-N-MNB carbonylation, (5) HPLC analysis of the mono-N-MNB carbonylated derivatives.
The so-called "new generation reagent" (^+)-MNB carboxylic acid was superior in the D,L-analysis of amino alcohols than the "old" (^+)-TBMB carboxylic acid. However, the mono-N-MNB-carbonylation method needed the authentic samples for unequivocal identification. Therefore, we next studied the exciton-CD monitoring method by N,O-di-O- (S)-TBMB-carbonylated amino alcohols were clearly separated by normal phase column chromatography but their stereochemistry could not be determined by CD monitoring because they did not show the characteristic exciton CD spectra for the specific stereoisomer. Finally, we have found that the amino alcohols can be determined by the on-line HPLC-exciton CD analysis without reference asmples by N-phthaloyl-O-(S)-TBMB-carbonylation. Thus, we have developed a new sensitive and selective method for the determination of D,L of amino acids.
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