Zhu Yingying, Chen Roulin, Wang Hao, Chen Yihan, Liu Yuanlin, Zhou Jingwen, Mu Wanmeng
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
Bloomage Biotechnology Corporation, Limited, Jinan, Shandong 250010, People's Republic of China.
J Agric Food Chem. 2023 Mar 29;71(12):4915-4923. doi: 10.1021/acs.jafc.3c00139. Epub 2023 Mar 6.
2'-Fucosyllactose (2'-FL) is a kind of fucosylated human milk oligosaccharide (HMO), representing the most abundant oligosaccharide in breast milk. We conducted systematic studies on three canonical α1,2-fucosyltransferases (WbgL, FucT2, and WcfB) to quantify the byproducts in a - and -deleted BL21(DE3) basic host strain. Further, we screened a highly active α1,2-fucosyltransferase from sp. 11S02629-2 (BKHT), which exhibits high 2'-FL productivity without the formation of byproducts difucosyl lactose (DFL) and 3-FL. The maximum 2'-FL titer and yield reached 11.13 g/L and 0.98 mol/mol of lactose, respectively, in shake-flask cultivation, both approaching the theoretical maximum value. In a 5 L fed-batch cultivation, the maximum 2'-FL titer reached 94.7 g/L extracellularly with a yield of 0.98 mol of 2'-FL/mol of lactose and productivity of 1.14 g L h. Our reported 2'-FL yield is the highest from lactose reported to date.
2'-岩藻糖基乳糖（2'-FL）是一种岩藻糖基化的人乳寡糖（HMO），是母乳中含量最丰富的寡糖。我们对三种典型的α1,2-岩藻糖基转移酶（WbgL、FucT2和WcfB）进行了系统研究，以定量在缺失α和β的BL21(DE3)基础宿主菌株中的副产物。此外，我们从嗜热栖热放线菌11S02629-2（BKHT）中筛选出一种高活性的α1,2-岩藻糖基转移酶，该酶具有高2'-FL生产能力，且不形成副产物二岩藻糖基乳糖（DFL）和3-FL。在摇瓶培养中，2'-FL的最大滴度和产量分别达到11.13 g/L和0.98 mol/mol乳糖，均接近理论最大值。在5 L补料分批培养中，细胞外2'-FL的最大滴度达到94.7 g/L，产量为0.98 mol 2'-FL/mol乳糖，生产效率为1.14 g L h。我们报道的2'-FL产量是迄今为止以乳糖为原料所报道的最高产量。
