Biomineralization-driven one-pot separation and stabilization of extracellular lipases bypassing purification

The industrial applications of enzymes are often hindered by labor-intensive purification and immobilization processes, which reduce enzyme activity and limit scalability. To overcome these limitations, we developed a one-pot calcium-based biomineralization strategy that enables the direct separation and immobilization of extracellular lipases secreted by Magnusiomyces sp. LA-1, without prior purification. During the biomimetic process, in situ precipitation of calcium carbonate microbeads entrapped secreted lipases (CaCO₃@MLALPs) under mild aqueous conditions, maintaining their native conformation and catalytic functionality. The biomineralized lipases exhibited approximately seven-fold higher hydrolytic activity and 1.6-fold greater esterification efficiency than the purified enzyme, indicating that the mineral matrix provided both structural rigidity and molecular stabilization. The CaCO₃@MLALPs retained 46 % of their activity after nine reuse cycles and 40 % after 100 d of storage, demonstrating excellent long-term durability and reusability. Under the esterification conditions, a glyceryl caprylate yield of 253 mM was achieved, confirming its practical potential in synthetic bioprocesses. Overall, this study developed a cost-effective and sustainable biomineralization platform for the direct recovery of extracellular enzymes, highlighting a purification-free and scalable approach to enzyme immobilization with enhanced stability and catalytic performance.