Rebaudioside A was modified via glucosylation by recombinant dextransucrase of Leuconostoc lactis EG001 in Escherichia coli BL21 (DE3), forming single O‐α‐D‐glucosyl‐(1″→6′) rebaudioside A with yield of 86%. O‐α‐D‐glucosyl‐(1″→6′) rebaudioside A was purified using HPLC and Diaion HP‐20 and its properties were characterized for possible use as a food ingredient. Almost 98% of O‐α‐D‐glucosyl‐(1″→6′) rebaudioside A was dissolved after 15 days of storage at room temperature, compared to only 11% for rebaudioside A. Compared to rebaudioside A, O‐α‐D‐glucosyl‐(1″→6′) rebaudioside A showed similar or improved acidic or thermal stability in commercial drinks. Thus, O‐α‐D‐glucosyl‐(1″→6′) rebaudioside A could be used as a highly pure and improved sweetener with high stability in commercial drinks. Practical Application The proposed method can be used to generate glucosyl rebaudioside A by enzymatic glucosylation. Simple glucosyl rebaudioside A exhibited high acid/thermal stability and improved sweetener in commercialized drinks. This method can be applied to obtain high value‐added bioactive compounds by enzymatic modification.

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