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Laccase producted by this fungus in cultures supplemented with copper sulfate was purified 60-fold by ammonium-sulfate precipitation, Sephadex G-100 gel-filtration chromatography, DEAE Sepharose ion-exchange chromatography, and Superdex G-75 gel-filtration chromatograpgy, with a yield of 3.3%. Native and SDS-PAGE showed an active band with a molecular mass of approximately 67,000. Purified laccase maintained 70-80% stability at pH 6-10 for 180 min and at 30-40°C for 120 min. The optimum pH for the oxidation of 2,2´-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was pH 3, while that for the oxidation of 2,6-dimethoxyphenol (DMP), guaiacol, and syringaldazine was pH 6. Kinetic constants (Km and kcat/km) showed that 2,6-DMP and syringaldazine were more effective substrates than ABTS and guaiacol. Enzyme activity was completely inhibited by sodium azide or L-cysteine at a concerntration of 1 mM. Laccase activity was also inhibited by 5 mM SDS.\\\n[要約]ブナ材に軟腐朽を起こすGraphiumsp. 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〈Originals〉Purification and partial characterization of laccase from the soft-rot fungus Graphium sp. M-1-9a
https://kindai.repo.nii.ac.jp/records/19191
https://kindai.repo.nii.ac.jp/records/19191f6bf6949-fed1-4c95-abf4-7bc2406db9d1
名前 / ファイル | ライセンス | アクション |
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AN00064044-20180331-0035.pdf (460.1 kB)
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Item type | ☆紀要論文 / Departmental Bulletin Paper(1) | |||||
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公開日 | 2018-03-08 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | 〈Originals〉Purification and partial characterization of laccase from the soft-rot fungus Graphium sp. M-1-9a | |||||
その他(別言語等)のタイトル | ||||||
その他のタイトル | 〈原著〉軟腐朽を起こすGraphium sp. M-1-9のラッカーゼの諸性質について | |||||
著者 |
田中, 裕美
× 田中, 裕美× 山本, 貴義× 本田, 克樹× 板倉, 修司 |
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言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題 | soft-rot fungus, laccase, Graphium sp., wood degradation | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | departmental bulletin paper | |||||
著者(英) | ||||||
en | ||||||
TANAKA,Hiromi | ||||||
著者(英) | ||||||
en | ||||||
YAMAMOTO,Takanori | ||||||
著者(英) | ||||||
en | ||||||
Honda,Katsuki | ||||||
著者(英) | ||||||
en | ||||||
ITAKURA,Shuji | ||||||
著者 所属 | ||||||
近畿大学農学部; 名誉教授 | ||||||
著者 所属 | ||||||
近畿大学大学院農学研究科応用生命化学専攻 | ||||||
著者 所属 | ||||||
近畿大学大学院農学研究科応用生命化学専攻 | ||||||
著者 所属 | ||||||
近畿大学大学院農学研究科応用生命化学専攻 | ||||||
著者所属(翻訳) | ||||||
Kindai University | ||||||
著者所属(翻訳) | ||||||
Kindai University | ||||||
著者所属(翻訳) | ||||||
Kindai University | ||||||
著者所属(翻訳) | ||||||
Kindai University | ||||||
版 | ||||||
出版タイプ | NA | |||||
出版タイプResource | http://purl.org/coar/version/c_be7fb7dd8ff6fe43 | |||||
出版者 名前 | ||||||
出版者 | 近畿大学農学部 | |||||
書誌情報 |
近畿大学農学部紀要 en : MEMOIRS OF THE FACULTY OF AGRICULTURE OF KINDAI UNIVERSITY 号 51, p. 35-50, 発行日 2018-03-31 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 21896267 | |||||
抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | [Synopsis]An extracellular laccase was isolated, purified, and partially characterized from the culture medium of Grphium sp. M-1-9, which causes soft rot in Japanese beech wood. Laccase producted by this fungus in cultures supplemented with copper sulfate was purified 60-fold by ammonium-sulfate precipitation, Sephadex G-100 gel-filtration chromatography, DEAE Sepharose ion-exchange chromatography, and Superdex G-75 gel-filtration chromatograpgy, with a yield of 3.3%. Native and SDS-PAGE showed an active band with a molecular mass of approximately 67,000. Purified laccase maintained 70-80% stability at pH 6-10 for 180 min and at 30-40°C for 120 min. The optimum pH for the oxidation of 2,2´-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was pH 3, while that for the oxidation of 2,6-dimethoxyphenol (DMP), guaiacol, and syringaldazine was pH 6. Kinetic constants (Km and kcat/km) showed that 2,6-DMP and syringaldazine were more effective substrates than ABTS and guaiacol. Enzyme activity was completely inhibited by sodium azide or L-cysteine at a concerntration of 1 mM. Laccase activity was also inhibited by 5 mM SDS.\ [要約]ブナ材に軟腐朽を起こすGraphiumsp. M-1-9を、硫酸銅を添加したブナ木片培地で培養し、ラッカーゼを分離・精製し、その諸性質を検討した。硫安沈殿、ゲルロ過クロマトグラフィー、イオン交換クロマトグラフィーにより60倍に精製されたラッカーゼの収量は3.3%であった。SDS-PAGEにより、分子量は約67000であった。精製したラッカーゼはpH 6-10では180分間、また30-40℃では120分間、70-80%の活性を維持した。最適pHは基質により異なっており、2,2ʹ-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS)ではpH 3、2,6-dimethoxyphenol(DMP)、guaiacol,、syringaldazineではpH 6であった。Kmとkcat/Kmより2,6-DMP とsyringaldazineは、ABTS とguaiacolより効率的な基質であった。酵素活性は1 mM.のアジ化ナトリウムあるいはL-システイン、5 mM SDSで阻害された。 |
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フォーマット | ||||||
内容記述タイプ | Other | |||||
内容記述 | application/pdf |