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Microbiology and Biotechnology Letters

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Microbiol. Biotechnol. Lett. 2009; 37(3): 204-212

Weissella cibaria가 생산하는${alpha}$-Galactosidase 및 ${beta}$-Glucosidase의 특성

Characterization of ${alpha}$-Galactosidase and ${beta}$-Glucosidase by Weissella cibaria

Sung-Wook Hong , Lae-Kyun You , Byung-Moon Jung , Wan-Sik Kim and Kun-Sub Chung

Division of Biological Science and Technology, Yonsei University, 1Division of Biological Science and Technology, Yonsei University, 2R&D Center, Maeil Dairies Co. Ltd., 3R&D Center, Maeil Dairies Co. Ltd., 4Division of Biological Science and Technology, Yonsei University

Abstract

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A strain producing ${alpha}$-galactosidase and ${beta}$-glucosidase was isolated from Kimchi. The isolated strain was identified as Weissella cibaria by 16S rDNA analysis and designated as Weissella cibaria K-M1-4. The enzyme activity of ${alpha}$-galactosidase and ${beta}$-glucosidase reached the maximum in the soy medium at $37^{circ}C$ for 24 hr. The enzymes were purified by ethanol fractionation, DEAE sepharose fast flow, and sephacryl S-100HR column chromatography. ${alpha}$-Galactosidase specific activity was shown by 576 Units/mg protein and the yield was 3.5% of the total activity of crude extracts. ${beta}$-glucosidase specific activity was shown by 480 Units/mg protein and the yield was 2.9% of the total activity of crude extracts. The optimum temperature for ${alpha}$-galactosidase was $60^{circ}C$ and 43% of its original activity remained when it was treated at $80^{circ}C$ for 30 min. For ${alpha}$-galactosidase shows the optimum pH of 8.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 0.98 mM and $1.81{mu}$mole/min, respectively. The ${beta}$-glucosidase has the optimum temperature of $50^{circ}C$ and 46% of its original activity remained when it was treated at $80^{circ}C$ for 30min. Its optimum pH of 7.0 and is fairly stable between pH5.0 and pH9.0. The enzyme activity was increased in the presence of $Fe^{2+},;Co^{2+}$ and $Cu^{2+}$. The value of Km and Vmax for the enzyme were 1.24 mM and $6.81{mu}$mole/min, respectively.

Keywords: Kimchi, Weissella cibaria, ${alpha}$-galactosidase, ${beta}$-glucosidase

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