Key Laboratory of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, National Engineering Laboratory for Industrial Enzymes, Tianjin 300457, PR China.
Key Laboratory of Industrial Fermentation Microbiology of the Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science and Technology, National Engineering Laboratory for Industrial Enzymes, Tianjin 300457, PR China.
Food Chem. 2023 Feb 1;401:134199. doi: 10.1016/j.foodchem.2022.134199. Epub 2022 Sep 12.
D-Allulose, as low-calorie rare sugar, possessed several notable biological activities and was biosynthesized by D-allulose 3-epimerase (DAEase). Here, CcDAE from Clostridium cellulolyticum was successfully immobilization via covalent attachment (RI-CcDAE), and Resin-SpyCatcher/SpyTag-CcDAE modular (DI-CcDAE). Both immobilized CcDAEs exhibited higher thermal and pH stabilities than the free form, and they maintained 80.0 % of relative activity after 7 consecutive cycles and 25 days of storage. Predominantly, DI-CcDAE represented superior catalytic efficiency with a 2.4-fold increase of k/K, compared with RI-CcDAE (0.75 s mM vs 0.31 s mM). The RI-CcDAE and DI-CcDAE were then applied in mixed fruit Jiaosu to convert D-fructose into D-allulose, which exhibited the productivity of D-allulose 1.08 g/Lh and 1.57 g/Lh, respectively. This research provided a promising directional immobilization strategy for DAEase, and robust biocatalyst for production of functional foodstuff containing D-allulose.
D-阿洛酮糖作为一种低热量稀有糖，具有多种显著的生物学活性，可通过 D-阿洛酮糖 3-差向异构酶（DAEase）生物合成。本文通过共价附着（RI-CcDAE）和树脂-SpyCatcher/SpyTag-CcDAE 模块（DI-CcDAE）成功实现了来自纤维丁酸弧菌的 CcDAE 的固定化。与游离形式相比，两种固定化的 CcDAE 均表现出更高的热稳定性和 pH 稳定性，经过 7 次连续循环和 25 天储存后，相对活性仍保持在 80.0%。尤其是 DI-CcDAE 的催化效率更高，k/K 值增加了 2.4 倍，而 RI-CcDAE 为 0.75 s mM 和 0.31 s mM。然后将 RI-CcDAE 和 DI-CcDAE 应用于混合水果酱汁中，将 D-果糖转化为 D-阿洛酮糖，其产物的 D-阿洛酮糖得率分别为 1.08 g/Lh 和 1.57 g/Lh。本研究为 DAEase 的定向固定化提供了一种很有前途的策略，也为生产含有 D-阿洛酮糖的功能性食品提供了一种稳健的生物催化剂。
