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KMID : 0545120010110020242
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Journal of Microbiology and Biotechnology
2001 Volume.11 No. 2 p.242 ~ p.250
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Purification and Properties of Cyclodextrin Glucanotransferase Synthesizing 2-O-α-D-Glucopyranosyl L-Ascorbic Acid from Paenibacillus sp. JB-13
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Bae, Kyung Mi
Kim, Sung Koo/Kong, In Soo/Jun, Hong Ki
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Abstract
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A Gram-positive bacterium (strain JB-13) that was isolated from soil as a producer of cyclodextrin glucanotransferase (CGTase) [EC 2.4.1.19] was identified as Paenibacillus sp. JB-13. This CGTase could catalyze the transglucosylation reaction from soluble starch to L-ascorbic acid (AA). A main product formed by this enzyme with α-glucosidase was identified as 2-O-α-D-glucopyranosyl L-ascorbic acid (AA-2G) by the HPLC profile and the elemental analysis. CGTase was purified to homogeneity using ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-50, and gel chromatography on Sephacryl S-200HR. The molecular weight was determined to be 66,000 by both gel chromatography and SDS-PAGE. The isoelectric point of the purified enzyme was 5.3. The optimum pH and temperature was pH 7.0 and 45℃, respectively. The enzyme was stable in the range of pH 6-9 and at temperatures of 75℃ or less in the presence of 15mM CaCl_2. Hg^2+, Mn^2+, Ag^+, and Cu^2+ all strongly inhibited the enzyme´s activity.
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