Characterization and elucidation of a novel M-specific alginate lyase Aly7Aq with strict recognition at subsites ±2

• Published in: International journal of biological macromolecules 2024-10, Vol.277 (Pt 1), p.133972, Article 133972
• Main Authors: Li, Jiajing, Sun, Menghui, Liu, Guanchen, Zhou, Jinhang, Chang, Yaoguang, Xue, Changhu
• Format: Article
• Language: English
• Subjects: Alginate lyase; M-specific; Oligosaccharide products
• ISSN: 0141-8130; 1879-0003; 1879-0003
• DOI: 10.1016/j.ijbiomac.2024.133972

A novel alginate lyase Aly7Aq was cloned and heterologous expressed by a combination of bioinformatics and molecular biology. Aly7Aq was an M-specific alginate lyase, exhibiting optimum reaction conditions at 50 °C and pH 10.0. Aly7Aq was determined to degrade polysaccharides in a random endo-acting manner. The minimum reaction substrate was tetrasaccharide, and Aly7Aq mainly attacked the third glycosidic linkage from the reducing end of oligosaccharide substrates. The disaccharide product of Aly7Aq was ΔM and the trisaccharide products were ΔMM and ΔMG, which differed from all previously characterized M-specific alginate lyases. The degradation products demonstrated that the ±2 subsites of Aly7Aq strictly recognized M units, while the −1 subsite accommodated both M and G units. Therefore, the substrate specificity of Aly7Aq was derived from the specificity of ±2 subsites. This is the first report on the specificity at subsite ±2 of M-specific alginate lyase. The novel M-specific Aly7Aq could serve as a potential tool in the specific degradation of alginate and targeted preparation of oligosaccharide. •A novel M-specific alginate lyase Aly7Aq was cloned, expressed and characterized.•Aly7Aq chiefly cut the third glycosidic linkage from the reducing end of oligomannuronates.•The unique products of Aly7Aq were showed by HPAEC-PAD/MS with the trisaccharide products ΔMM and ΔMG.•The substrate specificity of Aly7Aq was derived from the strict recognition of its ±2 subsites.