Significant components of cost-effective medium for Magnusiomyces capitatus A4C extracellular lipase (ECL) production were optimized via a five-level factorial design. A simplistic, economical, and green approach was adopted for biomimetic mineralization to prepare multilayered nano-entrapped ECL, which were then applied as biocatalysts for the production of fatty acid methyl ester (FAME). The optimal ECL (0.8 mg protein/mL) and CuSO4 center dot 5H(2)O (1.2 mM) showed the highest capacity for enzyme loading. The ECL-CuSO4-hybrid showed an 89.7% conversion of triacylglycerides into FAME via transesterification and a 98.7% conversion of oleic acid into FAME via esterification at 72 h. The ECL-CuSO4-hybrid gave 65% and 78.7% FAME production after 5 successive reuses via transesterification and esterification reactions, respectively. Therefore, these ECL-inorganic hybrid biocatalysts have high economical potential to be used for the production of biodiesel as the future petrodiesel replacement.