利用1-2-岩藻糖基转移酶进行化学酶法合成岩藻糖基化寡糖及其在肠道微生物群调节中的应用

Chemoenzymatic synthesis of fucosylated oligosaccharides using 1-2-fucosyltransferase and their application in the regulation of intestinal microbiota.

作者信息

Zhong Ruting, Gao Luying, Chen Zhengxin, Yuan Sinan, Chen Xi, Zhao Chao

机构信息

Engineering Research Centre of Fujian-Taiwan Special Marine Food Processing and Nutrition, Ministry of Education, Fuzhou 350002, China.

College of Food Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China.

出版信息

Food Chem X. 2021 Nov 2;12:100152. doi: 10.1016/j.fochx.2021.100152. eCollection 2021 Dec 30.

A novel bacterial 1-2-fucosyltransferase ( 2FT) from sp. NK55a (Ts2FT) has been discovered and characterized. It shares 28-62% protein sequence homology to 2FTs reported previously. The Ts2FT was cloned as an -terminal His-tagged recombinant protein (His-Ts2FT) and expressed in BL21 (DE3). It was expressed at a level of 6.2 mg/L culture after induction with 0.05 mM of isopropylβ-d-1-thiogalactoside (IPTG) at 16 °C for 20 h. It showed the optimal activity at a reaction temperature of 40 °C and pH of 7.0. The presence of a Mg improved its catalytic efficiency. Ts2FT displayed a strict acceptor specificity and could recognize only β1-3-galatoside acceptors. It was used efficiently for one-pot multienzyme synthesis of fucosylated oligosaccharides. One of the products, lacto--fucopentaose I was shown to promote the growth of intestinal probiotics including those belonging to Acidobacteria, Actinobacteria, Proteobacteria, Planctomycetes, and Chloroflexi.

已发现并鉴定了一种来自sp. NK55a的新型细菌1-2-岩藻糖基转移酶（2FT，即Ts2FT）。它与先前报道的2FTs的蛋白质序列同源性为28%-62%。Ts2FT被克隆为N端带有His标签的重组蛋白（His-Ts2FT），并在BL21(DE3)中表达。在16℃用0.05 mM异丙基-β-D-1-硫代半乳糖苷（IPTG）诱导20小时后，其表达水平为6.2 mg/L培养物。它在40℃的反应温度和pH 7.0时表现出最佳活性。Mg的存在提高了其催化效率。Ts2FT表现出严格的受体特异性，只能识别β1-3-半乳糖苷受体。它被有效地用于一锅多酶合成岩藻糖基化低聚糖。其中一种产物，乳糖-N-岩藻五糖I被证明能促进包括属于酸杆菌、放线菌、变形菌、浮霉菌和绿弯菌的肠道益生菌的生长。