College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China.
College of Food Science and Engineering, Ocean University of China, Qingdao 266404, China; Laboratory for Marine Drugs and Bioproducts, Pilot National Laboratory for Marine Science and Technology, Qingdao 266237, China.
Int J Biol Macromol. 2023 May 31;238:124037. doi: 10.1016/j.ijbiomac.2023.124037. Epub 2023 Mar 15.
Sulfated fucan is an important functional polysaccharide with various physiological activities. Carbohydrate-binding module (CBM) is a representative class of carbohydrate-binding protein, which could be employed as a favorable tool for the investigations and applications of polysaccharides. Nevertheless, only one confirmed sulfated fucan-binding CBM has been hitherto reported. In the present study, an unknown domain with a predicted β-sandwich fold was discovered from a fucanase Fun174A, and further cloned and heterologously expressed in Escherichia coli. The expressed protein Fun174A-CBM displayed a specific binding capacity to sulfated fucan. The bio-layer interferometry assays showed that the protein could bind to the sulfated fucan tetrasaccharide with an affinity constant of 2.83 × 10 M. Fun174A-CBM shared no significant sequence similarity to any identified CBMs, indicating that it represents a new CBM family. The discovery of Fun174-CBM and the novel CBM family would be beneficial to the investigations of sulfated fucan-binding proteins.
硫酸化岩藻聚糖是一种具有多种生理活性的重要功能多糖。糖结合模块(CBM)是糖结合蛋白的一个代表性类别,可作为多糖研究和应用的有利工具。然而,迄今为止仅报道了一种已确认的硫酸化岩藻聚糖结合 CBM。在本研究中,从岩藻聚糖酶 Fun174A 中发现了一个具有预测β-三明治折叠的未知结构域,并进一步在大肠杆菌中克隆和异源表达。表达的蛋白 Fun174A-CBM 显示出对硫酸化岩藻聚糖的特异性结合能力。生物层干涉测定法显示,该蛋白可以与硫酸化岩藻四糖结合,亲和力常数为 2.83×10^-5 M。Fun174A-CBM 与任何已鉴定的 CBM 均无明显的序列相似性,表明它代表了一个新的 CBM 家族。Fun174-CBM 的发现和新型 CBM 家族将有助于硫酸化岩藻聚糖结合蛋白的研究。