Purification and Some Properties of a β-Glucosidase from Flavobacterium johnsonae
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- OKAMOTO Katsuyuki
- Showa Sangyo Co., Ltd.
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- NAKANO Hirofumi
- Osaka Municipal Technical Research Inst.
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- YATAKE Tsunneya
- Showa Sangyo Co., Ltd.
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- KISO Taro
- Osaka Municipal Technical Research Inst.
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- KITAHATA Sumio
- Osaka Municipal Technical Research Inst.
書誌事項
- タイトル別名
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- Purification and Some Properties of a .BETA.-Glucosidase from Flavobacterium johnsonae.
- Purification and Some Properties of a ベータ Glucosidase from Flavobacterium johnsonae
- Purification and Some Properties of a β-Glucosidase from<i>Flavobacterium johnsonae</i>
この論文をさがす
抄録
Flavobacterium johnsonae was isolated as a microorganism that produced a β-glucosidase with hydrolytic activity of β-glucosyl ester linkages in steviol glycosides. The enzyme was purified to homogeneity from a cell-free extract by streptomycin treatment, ammonium sulfate fractionation, and column chromatographies on S-Sepharose and phenyl-Toyopearl. The molecular mass of the purified enzyme was about 72 kDa by SDS-PAGE. An isoelectric point of pI 8.8 was estimated by isoelectric focusing. The enzyme was most active at pH 7.0, and was stable between pH 3.0 and 9.0. The optimum temperature was 45°C, and the enzyme was stable below 35°C. The enzyme hydrolyzed glucosyl ester linkages at site 19 of rebaudioside A, stevioside, and rubusoside, although it could not degrad β-glucosidic linkages at site 13 of rebaudioside B or steviol bioside. The enzyme acted on aryl β-glucosides such as p-nitrophenyl β-glucoside, phenyl β-glucoside, and salicin, and glucobioses such as sophorose and laminaribiose. The enzyme activity on Rub was inactivated completely by Hg2+, and reduced by Fe3+, Cu2+, p-chloromercuric benzoate, and phenylmethylsulfonyl fluoride (residual activity; 67.9-84.8%). The pNPG hydrolysis was also inactivated to almost the same degrees. Kinetic behaviors in the mixed substrate reactions of rebaudioside A and steviol monoside, and of steviol monoglucosyl ester and phenyl β-glucoside suggested the glucosidic and glucosyl ester linkages were hydrolyzed at a single active site of the enzyme.<br>
収録刊行物
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 64 (2), 333-340, 2000
公益社団法人 日本農芸化学会
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詳細情報 詳細情報について
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- CRID
- 1390282681449894400
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- NII論文ID
- 110002679914
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- NII書誌ID
- AA10824164
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- COI
- 1:CAS:528:DC%2BD3cXhvVCjt7w%3D
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- ISSN
- 13476947
- 09168451
- http://id.crossref.org/issn/09168451
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- NDL書誌ID
- 5296349
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- PubMed
- 10737190
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- 抄録ライセンスフラグ
- 使用不可