Investigation of the Contribution of Late Embryogenesis Abundant (LEA) K Peptide in Enhancing the Expression of Lipase from Sphingobacterium sp.: In vitro and in Silico Studies

Abstract

Sphingobacterium sp. AB3 lipase is a psychrophilic enzyme with optimal lipolytic activity at pH 7 and 15°C. The cold-adapted properties of the lipase render it suitable for various low-temperature industrial applications such as biodiesel production, detergent formulation, and non-thermal food processing. Recent studies have shown that co-expression of LEA K peptide with AB3 lipase resulted in enhanced protein expression in Escherichia coli. In this study, the purified AB3 lipase was characterized by substrate specificity, followed by tertiary structure prediction of the lipase and LEA K peptide using SWISS-MODEL and PEP-FOLD 3.5, respectively. Molecular docking studies were conducted to study the lipase-LEA K interactions using ClusPro and lipase-olive oil interactions with and without LEA K using Autodock Vina. Based on the findings, AB3 lipase showed the highest preference for olive oil with a lipase-specific activity of 153.3 U/mg. In the presence of LEA K, the binding affinity of AB3 lipase with olive oil improved from -4.7 to -7.0, kcal/moL with increased hydrophobic interactions and hydrogen bonding with catalytic residues of the lipase. Overall, understanding the interaction between the AB3 lipase and LEA K peptide offers valuable insights into the mechanisms underlying the improved stability and affinity of the protein-peptide complex.