Fucosylated oligosaccharides present in human milk perform various biological functions that benefit infants’ health. These compounds can be also obtained by enzymatic synthesis. In this work, the effect of the immobilization of α-l-fucosidase from Thermotoga maritima on the synthesis of fucosylated oligosaccharides was studied, using lactose and 4-nitrophenyl-α-l-fucopyranoside (pNP-Fuc) as acceptor and donor substrates, respectively, and Eupergit^® CM as an immobilization support. The enzyme was immobilized with 90% efficiency at pH 8 and ionic strength of 1.5 M. Immobilization decreased enzyme affinity for the donor substrate as shown by a 1.5-times higher K_M value and a 22-times decrease of the k_cat/K_M ratio in comparison to the unbound enzyme. In contrast, no effect was observed on the synthesis/hydrolysis ratio (r_s/r_h) when α-l-fucosidase was immobilized. Also, the effect of initial concentration of substrates was studied. An increase of the acceptor concentration improved the yields of fucosylated oligosaccharides regardless enzyme immobilization. The synthesis yields of 38.9 and 40.6% were obtained using Eupergit^® CM-bound or unbound enzyme, respectively, and 3.5 mM pNP-Fuc and 146 mM lactose. In conclusion, α-l-fucosidase from Thermotoga maritima was efficiently immobilized on Eupergit^® CM support without affecting the synthesis of fucosylated oligosaccharides.

人乳中存在的岩藻糖基化低聚糖具有多种有益婴儿健康的生物学功能。这些化合物也可以通过酶促合成获得。在这项工作中，以乳糖和4-硝基苯基-α - 1-岩藻糖苷（p NP-Fuc）为受体和供体底物，研究了从滨海嗜热菌中固定化α- 1-岩藻糖苷酶对岩藻糖基化寡糖合成的影响。分别与Eupergit ^® CM作为固定载体。在pH 8和1.5 M的离子强度下，以90％的效率固定了酶。固定化降低了酶对供体底物的亲和力，如K _M高1.5倍所示与未结合的酶相比，k _cat / K _M值降低了22倍。相反，当固定α- 1-岩藻糖苷酶时，未观察到对合成/水解比（r _s / r _h）的影响。此外，还研究了底物初始浓度的影响。受体浓度的增加提高了岩藻糖基化寡糖的产率，而与酶的固定无关。使用Eupergit得到38.9和40.6％的产率合成^® CM结合或未结合的酶，分别和3.5毫的pNP-Fuc的和146 mM的乳糖。总之，α-升从-fucosidase海栖热袍被有效固定在Eupergit ^® CM支持而不影响岩藻糖化寡糖的合成。