Purification and Characterization of a Novel Sennoside-Hydrolyzing β-Glucosidase from Bifidobacterium Sp. Strain SEN, a Human Intestinal Anaerobe
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概要
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A novel β-glucosidase, which is inducible and capable of catalyzing the hydrolysis of sennosides, was purified from Bifidobacterium sp. strain SEN with Triton X-100 solubilization and DEAE-cellulose column chromatography, by which hydrolytic activities toward sennoside B, 4-methylumbelliferyl β-glucoside (MUG), and p-nitrophenyl β-glucoside (pNPG) were obtained together in the same eluted fractions. The activity was stable against detergents such as sodium dodecyl sulfate (SDS) and Triton X-100,but was denatured by SDS and β-mercaptoethanal when heated. The final preparation was shown to be nearly homogeneous on SDS-polycrylamide gel electrophoresis (PAGE) either after the enzyme was denatured or when it was not denatured. In the non-denaturing SDS-PAGE, a single protein band hydrolyzed MUG on the gel. In the denaturing SDS-PAGE, the subunit mass of the enzyme was estimated to be 110 kDa.The enzyme was optimally active at pH 6.0 for hydrolysis of sennoside B and MUG. K_m values for sennoside B and MUG are 0.94 and 0.53 mM, respectively. The enzyme also catalyzed the hydrolysis of pNPG, amygdalin, geniposide and salicin. It was less active against methyl β-glucoside and incapable of hydrolyzing cellobiose. The β-glucosidase activity was inhibited by deoxynojirimycin and p-chloromercuribenzenesulfonic acid, but was less susceptible to several metals (FeSO_4,ZnCl_2,and CuSO_4), and 5,5'-dithio-bis(2-nitrobenzoic acid).
- 社団法人日本薬学会の論文
- 1996-05-15
著者
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Kobashi K
Toyama Medical And Pharmaceutical Univ. Toyama Jpn
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Kobashi Kyoichi
Faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceutical University
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Yang Ling
中華人民共和国
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Yang Ling
Faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceutical University
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AKAO Teruaki
Faculty of Pharmaceutical Sciences, University of Toyama
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Yang Ling
Department Of Mathematics Suzhou University
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Akao Teruaki
Faculty Of Pharmaceutical Sciences University Of Toyama
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Akao Teruaki
Faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceutical University
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Akao Teruaki
Graduate School Of Medicine And Pharmaceutical Sciences Univ. Of Toyama
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Akao Teruaki
Department Of Enzymatic Physiology Faculty Of Pharmaceutical Sciences Toyama Medical And Pharmaceuti
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Hattori Masao
Research Institute For Wakan-yaku (traditional Sino-japanese Medicines) Toyama Medical And Pharmaceu
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Kobashi Kyoichi
Faculty Of Phamaceutical Sciences Toyama Medical And Pharmaceutical University:fuji Chemical Industr
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HATTORI MASAO
Research Institute for Wakan-Yaku (Oriental Medicines)
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