Efficient preparation of fatty acid methyl esters using liquid lipase from thermotolerant fungus Rasamsonia emersonii via synergistic strategies

Lipase-catalyzed biodiesel preparation in non-aqueous media has the advantages of reduced energy consumption, simplified recovery of fatty acid methyl esters (FAMEs), and minimized environment pollution, etc. To discover a novel thermophilic lipase, achieve its high-level expression, and develop it as an efficient liquid biocatalyst for FAMEs production via methanolysis of plant oils in non-aqueous media, a lipase gene from the thermophilic fungus Rasamsonia emersonii was cloned and efficiently expressed through synergistic strategies including computational codon optimization and increased gene dosage in the host genome. The resulting liquid lipase preparation was directly employed as a biocatalyst for FAMEs production. Under optimized reaction conditions, a conversion rate of approximately 95% was achieved. This study provides a feasible approach for the large-scale production of thermophilic lipase. The strategy of directly using of liquid lipase as a biocatalyst for FAMEs preparation substantially simplifies enzyme immobilization, regeneration, and product recovery processes, offering an efficient alternative for enzymatic FAMEs production.