State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
J Agric Food Chem. 2022 Jul 27;70(29):9017-9025. doi: 10.1021/acs.jafc.2c02484. Epub 2022 Jul 14.
2'-Fucosyllactose (2'-FL) is the most abundant oligosaccharide in human milk. In this study, a highly efficient biosynthetic route for 2'-FL production was designed the pathway of GDP-l-fucose using engineered BL21(DE3). Specifically, plasmid-based strains with previously deleted and were further reconstructed by introducing pathway genes and α1,2-fucosyltransferase-encoding to realize 2'-FL synthesis. The 2'-FL titer was enhanced to 3.92 g/L by further introducing and . Subsequently, the additional expression cassette was chromosomally integrated into locus to strengthen fucosylation reaction and a strong constitutive promoter (P) was used to replace the original promoters of and to improve 2'-FL synthesis. The maximal 2'-FL titer reached 9.06 and 79.23 g/L in shake-flask and fed-batch cultivation, respectively. The 2'-FL productivity reached 1.45 g/L/h, showing remarkable production potential in large-scale industrial application.
2'-岩藻糖基乳糖（2'-FL）是母乳中含量最丰富的寡糖。在本研究中，设计了一种高效的 2'-FL 生产生物合成途径，利用工程化的 BL21(DE3)利用 GDP-L-岩藻糖途径。具体来说，通过引入途径基因和编码α1,2-岩藻糖基转移酶的，进一步构建了先前缺失的和的基于质粒的菌株，以实现 2'-FL 的合成。通过进一步引入和，2'-FL 的产量提高到 3.92 g/L。随后，将额外的表达盒染色体整合到位点，以加强岩藻糖基化反应，并使用强组成型启动子（P）替代和的原始启动子，以提高 2'-FL 的合成。在摇瓶和分批补料培养中，2'-FL 的最高产量分别达到 9.06 和 79.23 g/L。2'-FL 的生产率达到 1.45 g/L/h，在大规模工业应用中显示出显著的生产潜力。
