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

• Published in: Marine drugs Vol. 18; no. 4; p. 189
• Main Authors: Wang, Zhi-Peng, Cao, Min, Li, Bing, Ji, Xiao-Feng, Zhang, Xin-Yue, Zhang, Yue-Qi, Wang, Hai-Ying
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.04.2020; MDPI
• Subjects: Algae; Alginate lyase; Alginates; Alginates - metabolism; Alginic acid; Amino acids; Bacteria; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bioinformatics; Biomass; Biostimulants; Cloning; Cloning, Molecular; Cold; Cold Temperature - adverse effects; cold-adapted; Degree of polymerization; Enzyme Assays; Enzyme Stability; Enzymes; Hydrogen-Ion Concentration; Industrial applications; Industrial Microbiology; Industrial production; Lyases; Molecular Weight; NaCl-independent; Oligosaccharides; Oligosaccharides - metabolism; pH effects; pH-stable; Phaeophyceae - chemistry; Phylogenetics; Polymerization; Polysaccharide-Lyases - chemistry; Polysaccharide-Lyases - genetics; Polysaccharide-Lyases - metabolism; Processing fishery products; Proteins; Salinity tolerance; Seaweed - chemistry; Seaweed meal; Seaweed processing; Seaweeds; Sodium chloride; Substrate Specificity; Vibrio; Vibrio - enzymology; Vibrio - genetics; Waterborne diseases; Yarrowia - genetics; alginate lyase; pH-stable; cold-adapted; NaCl-independent
• ISSN: 1660-3397; 1660-3397
• DOI: 10.3390/md18040189

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 Vibrio sp. W2 was screened. Then, the gene ALYW201 was cloned from Vibrio sp. W2 and expressed in a food-grade host, Yarrowia lipolytica. 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.