| Project/Area Number |
63570993
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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 |
Chemical pharmacy
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KANEDA Miyuki Hiroshima University, School of Medicine Associate Professor, 医学部, 助教授 (40019675)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Shoshiro Hiroshima University, Professor Emeritus, 名誉教授 (40013304)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1990: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1989: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1988: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Histidinomycin / Antifungal antibiotic / Peptide antibiotic / Histidine / Phytopathogenic fungi / Reverse-phase HPLC / FAB-MS / 2D NMR / 多孔性合成吸着剤 / 逆相中圧カラムクロマトグラフィ- / シアノプロピルシリカ / 逆相中圧力 / ラムクロマトグラフィー / HPLC / アミノ酸組成 / 分子量 / 分子式 |
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| Research Abstract |
Histidinomycin ( HM ) was first isolated from the broth filtrate of Streptomyces thermotolerance and is strongly active against few genera of phytopathogenic fungi.^<1)> However, the reported procedure^<1)> for purification contained very many and complicated steps. Furthermore, the reported chemical properties possessed many inconsistent characters. In this project, I carried out reexamination on the isolation and purification of HM with improved and facile methods and then performed characterization of the purified HM, which prove to be a new antifungal peptide antibiotic. The results are as follows:
[1] Improvement of isolation and purification method. (1) HM hydrochloride can be absorbed onto ion-exchange resin Dowex 50x4 and can be eluted with NH_4Cl solution. (2) Adsorption onto porous polymer Sepabeads SP207 and elution with 100 % MeOH. (3) Separation and purification by preparative reverse-phase HPLC (Inertsil ODS, phosphate buffer(ph 7.3) - MeOH 9:1). (4) Adsorption onto SEP-PAK c-18 and elution with 100 % MeOH(desaltation).
[2] Chemical and physico-chemical properties and structure elucidation of HN. (1) Molecular weight of HM has been established to be 643 by FAB-MS of HM-HCL. (2) It is suggested with a qualitative chemical test that HM has sulfur atom(s) in the molecule. (3) From the ^<13>C NMR spectra date, HM seems to consist of 24 carbons and no methyl groups. (4) From the HRFAB-MS date and the elemental analysis, the molecular formula of HM should be C_<24>H_<37>N_9O_6S_3. (5) On complete acid hydrolysis, HM gave three kinds of amino acids and one of them was assigned to be histidine. (6) HM-HCl can be crystallized from EtOH - water solution to give fine needles.
|) K. Ishimaru, et al. J. Antibiotics 36, 1644, 1983.
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