Laboratory of Enzyme Chemistry, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, 159, Prospect 100-let Vladivostoku, 690022, Vladivostok, Russia.
Laboratory of Enzyme Chemistry, G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far-Eastern Branch of the Russian Academy of Sciences, 159, Prospect 100-let Vladivostoku, 690022, Vladivostok, Russia.
Arch Biochem Biophys. 2022 Oct 15;728:109373. doi: 10.1016/j.abb.2022.109373. Epub 2022 Aug 5.
In present work we provide the bioinformatic and biochemical characterization of six α-L-fucosidases that belong to the 29 and 95 families of glycoside hydrolases (GH) from the fucoidan-degrading locus of the marine bacterium Wenyingzhuangia fucanilytica CZ1127. The fucosidases FucWf1, FucWf2, FucWf3 and FucWf6 are relegated to the subfamily A of the GH29 family. The fucosidase FucWf4 bears a distant resemblance to the GH29 and does not belong to either the GH29A or the GH29B subfamilies. Apparently, FucWf4 is the first representative of a new subfamily within the GH29 family of α-L-fucosidases. For the first time the specificity of fucosidases has been studied using a series of fucoidan-related sulfated oligosaccharides. Studied α-L-fucosidases are able to cleave l-fucose from sulfated fucooligosacchrides after their treatment with exo-sulfatases. All studied α-L-fucosidases are cleaving the α-1→3- and α-1→4-linked terminal l-fucose in sulfated fucooligosaccharides. However, only FucWf3 is able to cleave off an α-1→2-linked l-fucose. The fucosidase FucWf5 of the GH95 family is shown to have higher activity on fucoidans than fucosidases of the GH29 family. Supposedly, the α-l-fucosidase FucWf5 participates in fucoidan debranching. The obtained data indicate different roles of fucosidases of the GH29 and GH95 families in the process of fucoidan degradation by the marine bacteria W. fucanilytica CZ1127.
在本工作中，我们提供了属于糖苷水解酶（GH）29 家族和 95 家族的六种α-L-岩藻糖苷酶的生物信息学和生化特性，这些酶来自海洋细菌Wenyingzhuangia fucanilytica CZ1127 的岩藻聚糖降解基因座。岩藻糖苷酶 FucWf1、FucWf2、FucWf3 和 FucWf6 被归类于 GH29 家族的 A 亚家族。岩藻糖苷酶 FucWf4 与 GH29 具有较远的相似性，不属于 GH29A 或 GH29B 亚家族。显然，FucWf4 是 GH29 家族中α-L-岩藻糖苷酶新亚家族的第一个代表。我们首次使用一系列岩藻聚糖相关的硫酸化寡糖研究了岩藻糖苷酶的特异性。研究的α-L-岩藻糖苷酶在用外切硫酸酯酶处理后能够从硫酸化岩藻寡糖中切割 L-岩藻糖。所有研究的α-L-岩藻糖苷酶都能切割硫酸化岩藻寡糖中α-1→3-和α-1→4-连接的末端 L-岩藻糖。然而，只有 FucWf3 能够切割掉α-1→2-连接的 L-岩藻糖。GH95 家族的岩藻糖苷酶 FucWf5 被证明对岩藻聚糖的活性高于 GH29 家族的岩藻糖苷酶。推测α-L-岩藻糖苷酶 FucWf5 参与了岩藻聚糖的分支切割。获得的数据表明 GH29 和 GH95 家族的岩藻糖苷酶在海洋细菌 W. fucanilytica CZ1127 降解岩藻聚糖的过程中发挥不同的作用。
