Tan_2023_Mol.Catal_546_113271

Increasing environmental concerns and significant demand for industrial fuels necessitate the development of alternate and sustainable energy sources. Biodiesel is a renewable and environmentally friendly fuel produced by trans-esterifying a variety of feedstocks. Lipases are robust triacylglycerol ester hydrolases that catalyze hydrolysis, esterification, interesterification, transesterification, acylation, acidolysis, alcoholysis, aminolysis, and resolution of racemates. Although the lipase-assisted greener biosynthesis of biodiesel has numerous advantages over the traditional alkali-based process, low catalytic efficiency, marginal stability, and high cost of lipase enzymes limit its widespread industrial processability. Protein engineering methodologies such as directed evolution, semi-rational design, and rational design can effectively tailor or improve the biocatalytic characteristics of lipase enzymes used in biodiesel generation, such as thermostability, solvent tolerance, activity, and substrate specificity. These innovative techniques improved our ability to predict structure-function relationships, engineering qualities, computational tools for designing new biocatalysts, and functional screening to manipulate enzyme traits for application in prevalent industrial bioprocesses. Many recent studies have demonstrated improved lipase performance, such as activity, stability, and specificity via protein engineering. This review spotlights a current overview of lipase engineering at the molecular scale with robust biocatalytic properties for biodiesel synthesis.