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从 中鉴定和表征一种新型的α-L-岩藻糖苷酶及其在 2'-岩藻糖基乳糖生产中的应用。

Identification and Characterization of a Novel α-L-Fucosidase from and Its Application for Production of 2'-Fucosyllactose.

机构信息

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.

出版信息

Int J Mol Sci. 2023 Jul 17;24(14):11555. doi: 10.3390/ijms241411555.

Abstract

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.

摘要

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。

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