Biochemical and computational study of an alginate lyase produced by Pseudomonas aeruginosa strain S21

• Main Authors: Firoozeh Piroozmand, Parinaz Ghadam, Mahboobe Zarrabi, Ahya Abdi Ali
• Format: Article
• Language: English
• Published: Mashhad University of Medical Sciences 2020-04-01
• Series: Iranian Journal of Basic Medical Sciences
• Subjects: alginate; alginate lyase; computational biology; enzyme assay; pseudomonas aeruginosa; strain s21
• Online Access: http://ijbms.mums.ac.ir/article_14788_098ee43aa148f3558057f72d42b539c8.pdf

<em><strong>Objective(s):</strong></em> Alginates play a key role in mucoid Pseudomonas aeruginosa colonization, biofilm formation, and driving out of cationic antibiotics. P. aeruginosa alginate lyase (AlgL) is a periplasmic enzyme that is necessary for alginate synthesis and secretion. It also has a role in depolymerization of alginates. Using AlgLs in cystic fibrosis patients along with antibiotics enhances bacterial killing and host healing. In this study, we investigated the different biochemical properties of a newly isolated AlgL from P. aeruginosa S21 to complete the databank of AlgLs<br /><em><strong>Materials and Methods:</strong></em> The enzyme was extracted from the periplasmic space of the bacteria by the heat shock method. Using the TBA method, the enzyme activity and biochemical properties were assessed. The mutability of P. aeruginosa S21 AlgL to increase its thermal stability was investigated. The most favorable mutations were studied computationally. The molecular dynamics simulation (MDS) package GROMACS was used for determining the effect of S34R mutation on enzyme’s thermal stability.<br /><em><strong>Results:</strong></em> Data showed that this enzyme has the best activity at 37 °C and pH 7.5 and it can degrade mannuronate blocks, guluronate blocks, and sodium alginate. After 7 hr at 80 °C, 45% of the enzyme activity was retained. This enzyme needed 15 min to completely degrade accessible sodium alginate.  Tris buffer, pH 8.5 and Britton-Robinson buffer, pH 7.0 were the preferable buffers for the enzyme activity. MDS of native and mutated enzymes showed desirable results.<br /><em><strong>Conclusion:</strong></em> P. aeruginosa S21 AlgL can be used in medical and industrial applications to degrade alginates.