The Role of Surface Exposed Lysine in Conformational Stability and Functional Properties of Lipase from Staphylococcus Family

• Published in: Molecules (Basel, Switzerland) Vol. 25; no. 17; p. 3858
• Main Authors: Ahmad, Nurul Nadirah, Ahmad Kamarudin, Nor Hafizah, Leow, Adam Thean Chor, Rahman, Raja Noor Zaliha Raja Abd
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 25.08.2020; MDPI
• Subjects: Amino Acid Substitution; Amino acids; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; charged residue; Computer applications; Computer simulation; Dynamic stability; Enzyme Stability; Experimental methods; Exposure; Hot Temperature; Lipase; Lipase - chemistry; Lipase - genetics; Lysine; Lysine - chemistry; Lysine - genetics; mutagenesis; Mutants; Mutation; Mutation, Missense; Protein Domains; Proteins; Residues; Stability analysis; staphylococcal lipase; Staphylococcus - enzymology; Staphylococcus - genetics; Substrate preferences; Substrates; Surface charge; Surface stability; lysine; enzyme stability; charged residue; mutagenesis; staphylococcal lipase
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules25173858

Surface charge residues have been recognized as one of the stability determinants in protein. In this study, we sought to compare and analyse the stability and conformational dynamics of staphylococcal lipase mutants with surface lysine mutation using computational and experimental methods. Three highly mutable and exposed lysine residues (Lys91, Lys177, Lys325) were targeted to generate six mutant lipases . The model structures were simulated in water environment at 25 °C. Our simulations showed that the stability was compromised when Lys177 was substituted while mutation at position 91 and 325 improved the stability. To illustrate the putative alterations of enzyme stability in the stabilising mutants, we characterized single mutant K325G and double mutant K91A/K325G. Both mutants showed a 5 °C change in optimal temperature compared to their wild type. Single mutant K325G rendered a longer half-life at 25 °C (T = 21 h) while double mutant K91A/K325G retained only 40% of relative activity after 12 h incubation. The optimal pH for mutant K325G was shifted from 8 to 9 and similar substrate preference was observed for the wild type and two mutants. Our findings indicate that surface lysine mutation alters the enzymatic behaviour and, thus, rationalizes the functional effects of surface exposed lysine in conformational stability and activity of this lipase.