Purification, immobilization, evolution, and characterization of D-allulose 3-epimerase from Antarctic Microbacterium

文献类型： 外文期刊

第一作者： Li, Jingqi

作者： Li, Jingqi;Sun, Jingjing;Wang, Wei;Jiang, Chengcheng;Hao, Jianhua;Li, Jingqi;Hao, Jianhua;Hao, Jianhua

作者机构：

关键词： D-allulose; Biosynthesis; Immobilized enzyme

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）

ISSN： 0141-8130

年卷期： 2025 年 310 卷

页码：

收录情况： SCI

摘要： D-allulose is a rare, low-calorie sugar substitute with multiple physiological functions and is widely used in food and pharmaceutical industries. D-allulose is primarily produced by the catalytic conversion of D-fructose via allulose 3-epimerase (DAEase). In this study, a DAEase gene, dpema4, was isolated from an Antarctic bacterium and expressed in Escherichia coli. The recombinant DAEase was a homotetramer with an optimal temperature 60 degrees C and pH of 7.5. Its catalytic activity was not strictly dependent on metal ions, making it a safer alternative the other reported DAEases. The recombinant DAEase showed exhibited the highest activity towards D-allulose, and the bioconversion rate was 29 %. For immobilization, the cellulose-binding domain (CBD) was fused DAEase, and the fusion protein was immobilized on microcrystalline cellulose. The immobilized DAEase showed highly improved pH stability and maintained approximately 44 % of catalytic activity after 10 continuous action cycles. The single-point mutant A248H showed high thermal stability and catalytic activity at 60 degrees C, the bioconversion rate of D-fructose reached 32 %. In summary, the recombinant DAEase can serve as a good candidate enzyme for the production of D-allulose, and the establishment of a one-step purification immobilization of DAEase can facilitate its industrial application.

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