Main content area

Highly efficient biosynthesis of sucrose-6-acetate with cross-linked aggregates of Lipozyme TL 100 L

Yang, Xu’e, Zheng, Pu, Ni, Ye, Sun, Zhihao
Journal of biotechnology 2012 v.161 no.1 pp. 27-33
acetates, biosynthesis, coprecipitation, crosslinking, glutaraldehyde, immobilized enzymes, sucralose, sucrose, temperature, transesterification
As a short chain monoester, sucrose-6-acetate (S-6-a) is a key intermediate in the preparation of an eminent sweetener (sucralose). To replace the traditional multi-step chemical route for sucralose biosynthesis, enzymatic synthesis of S-6-a was investigated, using cross-linked enzyme aggregates (CLEAs) of Lipozyme TL 100 L. The optimal CLEA preparation conditions was obtained as follows: using 33.3% (v/v) PEG600 co-precipitated with additive of d-sorbierite, then cross-linking with 1.5% (v/v) glutaraldehyde at 0°C for 4h. As a result, the immobilized Lipozyme had high specific bioactivity (34.64U/g) of transesterification in non-aqueous media. With these immobilized enzymes, the optimum transesterification conditions were investigated systematically, including CLEA loading, the mole ratio of vinyl acetate versus sucrose, temperature and reaction time, etc. The results showed that the highest concentration and yield of S-6-a was 49.8g/L and 87.46%, respectively. Further experiments showed that the resulting CLEAs also had much higher operational stability than the commercial Lipozyme TLIM. The present work has paved a new path for the large-scale bioproduction of S-6-a with immobilized lipase in the future.