김치 시료로부터 자동산화되지 않으며 열 및 중성 pH에서 안정한 AA 유도체인 AA-2G를 생산할 수 있는 당전이활성을 가진 CGTase 생산균주를 분리하였고, 분리균주의 형태학적, 배양학적, 생리학적 성질 및 16s-rDNA sequences를 조사한 결과 그람 양서의 간균으로 호기성이며 내생포자를 형성하는 전형적인 중온성 Bacillus sp. JK-43으로 동정되었다. Bacillus sp. JK-43의 CGTase는 AA-2G 뿐만 아니라 AA-6G로 추정되는 물질을 함께 생산하였으며, 효소 최적생산조건은 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3;0.1%;K_2HPO_4,;그리고;0.02%;MgSO_4{cdot}7H_2O$가 함유된 배지에서 pH 7.0, $37^{circ}C$에서 26시간 동안 진탕배양하였을 때였다. 각종 당공여채에 따른 Bacillus sp. JK-43의 AA-2G 생산성을 조사한 결과 ${eta}-CD$에서 가장 높은 AA-2G 생산성을 보였으며, 식혜제도페액인 엿기름 및 밥당화액에서도 비교적 높은 AA-2G 생산성을 보였다. 또한 여러 가지 당수용체에 대한 JK-43의 CGTase의 당전이 반응을 검토한 결과 sucrose, mannitol 및 inositol에서 높은 당전이 수율인 70~90%를 나타내었다.
A bacterial strain, designated as JK-43, producing extracellular cyclodextrin glucanotransferase (CGTase)[EC 2.4.1.19] was isolated from kimchi. The CGTase from isolated strain JK-43 showed the transglucosylation activity from soluble starch to L-ascorbic acid(AA) compared to those obtained from other strains. A main product formed by this reaction was identified as $2-O-{alpha}-glucopyranosyl$ L-ascorbic acid(AA-2G) by testing its susceptibility to ${alpha}-glucosidase$ hydrolysis, the HPLC profiles, and through the elementary analysis. the ${eta}-CD,;{gamma}-CD$, potato starch and corn starch were identified to be suitable glucosyl donor for transglucosylation reaction on AA by CGTase. Acceptor specificity on AA-2G production was examined by use of AA, Iso-AA and AA-2P. Transglucosylation was observed toward AA-2P as well as AA and Iso-AA. The microorganism isolated from kimchi was identified as a strain of Bacillus sp. JK-43 based on the morphological, cultural, biochemical characteristics and partial 16SrDNA sequence analysis. The maximal CGTase production was observed in a medium containing 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3;0.1%;K_2HPO_4,;and;0.02%;MgSO_4{cdot}7H_2O$ with initial pH 7.0. The strain was cultured at $37^{circ}C$ for 26 hrs with reciprocal shaking.
