Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Purification and Properties of Cyclodextrin Glucanotrnsferase Synthesizing $2-O-{\alpha}-D-Glucopyranosyl{\;}_{L}-Ascorbic$ Acid from Paenibacillus sp. JB-13

  • Bae, Kyung-Mi (Division of Biological Sciences, Pusan National University) ;
  • Kim, Sung-Koo (Department of Biotechnology and Bioengineering, Pukyung National University) ;
  • Kong, In-Soo (Department of Biotechnology and Bioengineering, Pukyung National University) ;
  • Jun, Hong-Ki (Division of Biological Sciences, Pusan National University)
  • Published : 2001.04.01
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Abstract

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 Panibacillus 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 ${\alpha}-glucosidase$ was identified as $2-O-{\alpha}-D-glucopyranosyl{\;}_{L}-ascorbic$ 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-Seohadex 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 $45^{\circ}C$ respectively. The enzyme was stable in the range of pH 6-9 and at temperatures of $75{\circ}C$ or less in the presence of 15 mM ${CaCl_2}.\;{Hg^2+},\;{Mn^+2},{Ag^+},\;and\;{Cu^2+}$ all strongly inhibited the enzyme's activity.

Keywords

References

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