College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, China.
School of Pharmaceutical Sciences, Nanjing Tech University, 30 Puzhunan Road, Nanjing 211816, China.
Int J Mol Sci. 2023 Jul 17;24(14):11555. doi: 10.3390/ijms241411555.
2'-fucosyllactose (2'FL) is an important nutrient in human milk that stimulates beneficial microbiota and prevents infection. α-L-fucosidase is a promising component for 2'FL synthesis. In this study, a soil-oriented α-L-fucosidase-producing strain from ZS1 was isolated. was employed as a host for cloning and expressing the α-L-fucosidase gene (). The EntFuc was predicted as a member of the GH29 family with a molecular mass of 58 kDa. The optimal pH and temperature for the activity of EntFuc were pH 7.0 and 30 °C, respectively. The enzyme exhibited a strictly specific activity for 4-Nitrophenyl-α-L-fucopyranoside (pNP-Fuc) and had a negligible effect on hydrolyzing 2'FL. EntFuc could catalyze the synthesis of 2'FL via transfucosylation action from pNP-Fuc and lactose. The yield of 2'FL reached 35% under optimal conditions. This study indicated that EntFuc with a high conversion rate is a promising enzyme source for the biosynthesis of 2'FL.
2'-岩藻糖基乳糖(2'FL)是母乳中的一种重要营养物质,它能刺激有益菌群并预防感染。α-L-岩藻糖苷酶是合成 2'FL 的一种有前途的成分。本研究从 ZS1 中分离出一株以土壤为导向的α-L-岩藻糖苷酶产生菌。 被用作克隆和表达α-L-岩藻糖苷酶基因()的宿主。EntFuc 被预测为 GH29 家族的一员,分子量为 58 kDa。EntFuc 的最适 pH 和温度分别为 pH 7.0 和 30°C。该酶对 4-硝基苯基-α-L-岩藻吡喃糖苷(pNP-Fuc)具有严格的特异性活性,对水解 2'FL 的影响可以忽略不计。EntFuc 可以通过 pNP-Fuc 和乳糖的转糖基化作用催化 2'FL 的合成。在最佳条件下,2'FL 的产率达到 35%。本研究表明,具有高转化率的 EntFuc 是 2'FL 生物合成的一种有前途的酶源。