Cloning, Secretory Expression and Characterization of a Unique pH-Stable and Cold-Adapted Alginate Lyase

Abstract

Cold-adapted alginate lyases have unique advantages for alginate oligosaccharide (AOS) preparation and brown seaweed processing. Robust and cold-adapted alginate lyases are urgently needed for industrial applications. In this study, a cold-adapted alginate lyase-producing strain <i>Vibrio</i> sp. W2 was screened. Then, the gene <i>ALYW201</i> was cloned from <i>Vibrio sp.</i> W2 and expressed in a food-grade host, <i>Yarrowia lipolytica</i>. The secreted Alyw201 showed the activity of 64.2 U/mL, with a molecular weight of approximate 38.0 kDa, and a specific activity of 876.4 U/mg. Alyw201 performed the highest activity at 30 °C, and more than 80% activity at 25-40 °C. Furthermore, more than 70% of the activity was obtained in a broad pH range of 5.0-10.0. Alyw201 was also NaCl-independent and salt-tolerant. The degraded product was that of the oligosaccharides of DP (Degree of polymerization) 2-6. Due to its robustness and its unique pH-stable property, Alyw201 can be an efficient tool for industrial production.