Direct Influence of the Conserved Motif in PL7 Family Alginate Lyases on Enzyme Cold Adaptability

Abstract

Alginate lyase, a vital component of polysaccharide lyases, is instrumental in the efficient degradation of alginate and the production of single oligosaccharides. Although numerous alginate lyases have been characterized, only a few display extreme cold adaptability in the range of 0–20 °C. In this study, we identified a novel cold-adapted alginate lyase, Aly423, from Tamlana laminarinivorans PT2-4 isolated from Sargassum. Phylogenetic classification, enzyme structure, and catalytic property analyses confirmed that Aly423 could be classified as a member of subfamily 5 of the PL7 family and exhibited significant cold adaptability at low temperatures. Further analysis of the secondary structure and homology modeling of several cold-adapted enzymes revealed two variable amino acid sites in the conserved amino acid motif (YFK*G*Y) of Aly423, which may affect the cold adaptation mechanism. Point mutation experiments demonstrated that mutant A304T significantly altered the temperature adaptation of Aly423, highlighting the critical role of this amino acid site in the cold-adaptation mechanism of the enzyme. In summary, we effectively enhanced the enzymatic activity of the PL7 alginate cold-adapted enzyme through a rational design using computational methods. This advancement is of significant importance for the efficient utilization of sodium alginate in the food, agricultural, and pharmaceutical industries under low-temperature conditions.