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∙5H₂O (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.

通过五级因子设计优化了用于人头大孢菌A4C细胞外脂肪酶（ECL）生产的经济高效培养基的重要组成部分。采用一种简单，经济和绿色的方法进行仿生矿化，制备了多层纳米级包裹的ECL，然后将该ECL用作生产脂肪酸甲酯（FAME）的生物催化剂。最佳的ECL（0.8 mg蛋白/ mL）和CuSO ₄ ∙5H ₂ O（1.2 mM）显示出最高的酶负载能力。ECL-CuSO ₄杂化物在72小时内显示出通过酯交换反应将三酰基甘油酯转化为FAME的89.7％和通过酯化反应将油酸转化为FAME的98.7％。ECL-CuSO ₄经过酯交换和酯化反应连续5次重复使用后，β-杂化生成的FAME分别为65％和78.7％。因此，这些ECL-无机杂化生物催化剂具有很高的经济潜力，可用于生产生物柴油作为未来的石油柴油替代品。