Isolation of Aspergillus flavus MO-5 Producing Two Types of Intracellular α-D-Xylosidases : Purification and Characterization of α-D-Xylosidase I
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概要
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One strain (MO-5) of fungi producing α-D-xylosidase was isolated from sake koji by a method using glucose oxidase. MO-5 was identified as an Aspergillus flavus strain, produced no aflatoxin (B1, B2, G1, and G2). Two different types of α-D-xylosidases were detected in the cell-free extract. One of the enzymes (α-D-xylosidase I) was purified to an electrophoretically pure state by successive chromatography on Q-Sepharose, Phenyl Superose, PL-SAX, and TSK-gel G3000SW_XL. The purified enzyme hydrolyzed p-nitrophenyl α-D-xylopyranoside (α-p-NPX), methyl α-D-xylopyranoside, isoprimeverose [α-D-xylopy-ranosyl-(1→6)-D-glucopyranose], and xyloglucan oligosaccharide. The activity of this enzyme was highly specific for α-D-xylosidic linkages. The apparent K_m and V_<max> of the enzyme for α-p-NPX and isoprimeverose were 1.32 mM and 4.4 μmol/min/mg protein, and 4.00 mM and 58.8 μmol/min/mg protein, respectively. This enzyme had an apparent molecular weight of 100,000 by SDS-polyacrylamide gel electrophoresis and 400,000 by gel filtration chromatography (TSK-gel G3000SW_XL). The enzyme showed the highest activity at pH 4.5 and 45℃, and was stable from pH 4.5 to 6.0 and at temperatures up to 45℃. The activity was inhibited by SDS and Hg^<2+> , and slightly by Cu^<2+> and Fe^<3+>. This enzyme showed transfer action at high concentrations of isoprimeverose and transfer products were detected, and it had transxylosylation activity on maltose from isoprimeverose as a donor, too. From the results of the enzymatic digestion and the partial acidolysis of these prepared transfer products, their structures may be trisaccharides in which the xylosyl residues were transferred to isoprimeverose and maltose forming α-D-linkages. Accordingly, this enzyme was indicated to be able to transfer xylosyl residues to form α-D-xylosidic linkages.
- 社団法人日本農芸化学会の論文
- 1993-08-23
著者
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Yoshikawa K
Laboratory Of Food Microbiological Engineering Faculty Of Agriculture Kinki University
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Yoshikawa Kentaro
Laboratory Of Food Microbiological Engineering Faculty Of Agriculture Kinki University
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Yamamoto K
Laboratory Of Insect Genetic Resources Faculty Of Agriculture Kyushu University
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Yoshikawa K
Department Of Bioscience Faculty Of Bioresources Mie University
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Okada Shigetaka
Biochemical Research Laboratory, Ezaki Glico Co., Ltd.
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Yamamoto Kazuya
Biochemical Research Laboratory Ezaki Glico Co. Ltd.
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Yoshikawa Kenji
Biochemical Research Laboratory, Ezaki Glico Co., Ltd.
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Okada S
Biochemical Research Laboratory Ezaki Glico Co. Ltd.
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Okada Shigetaka
Biochemical Research Laboratory Ezaki Glico Co. Ltd.
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Yoshida Kouji
Department Of Apple Research National Institute Of Fruit Tree Science
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