GM3 trisaccharide biosynthesis and process optimization using engineered E. coli lysate and whole-cell catalysis
GM3 trisaccharide biosynthesis and process optimization using engineered E. coli lysate and whole-cell catalysis
复制标题
使用工程大肠杆菌裂解物和全细胞催化进行 GM3 三糖生物合成和工艺优化
DOI：
10.1080/07328303.2020.1788576
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发表时间：
2020
影响因子：
1
通讯作者：
Zhimeng Wu
中科院分区：
文献类型：
--
作者：
Lipeng Feng;Jie Shi;Haofei Hong;Zhifang Zhou;Zhimeng Wu
构建了同时编码CMP-Neu5Ac合成酶（SiaB）和α-2-3-唾液基转移酶（PmST3）的工程大肠杆菌BL21 (DE3、SiaB、PmST3、△lacZ、△nanZ)，合成GM3三糖。在最佳条件下，大肠杆菌裂解液可以很容易地用于GM3三糖的生物合成，产量接近定量。此外，在最佳条件下，该工程大肠杆菌菌株作为全细胞催化剂，在40ml培养基中可产生约1g的GM3三糖。该全细胞催化剂可循环使用至少3次，酶活性无明显损失。
An engineered E. coli strain BL21 (DE3, SiaB, PmST3, △lacZ, △nanZ) that simultaneously encoded CMP-Neu5Ac synthase (SiaB) and α-2-3-sialyltransferase (PmST3) was constructed to synthesize GM3 trisaccharide. The crude E. coli lysate could be readily used for biosynthesis of GM3 trisaccharide with a nearly quantitative yield under optimal conditions. In addition, this engineered E. coli strain as whole-cell catalyst could generate approximately 1 g of GM3 trisaccharide in 40 mL of medium under optimal conditions. The whole-cell catalyst could be recycled and reused for at least 3 rounds without obvious loss of enzymatic activity.