Biodiesel offers an important alternative to fossil fuel. In this work, Eversa Transform 2.0 lipase was immobilized onto 3-aminopropyltriethoxysilane (APTES) modified silica clay (SC) by glutaraldehyde. The characteristics of the functionalized supports and the immobilized lipase were investigated by FTIR, TEM, BET, and XRD. The results show that the optimal conditions of lipase immobilization are as follows: 2% glutaraldehyde concentration, 15 mg/mL lipase concentration and incubating at 25 °C for 60 min. The immobilized lipase showed a high tolerance to temperature and pH variation in comparison to the free lipase. The immobilized lipase on SC was applied as a biocatalyst for the synthesis of biodiesel from methanol and canola oil. A biodiesel yield of 86% was obtained at a temperature of 45 °C via a three-step methanol addition. A conversion yield of 67% was maintained after reusing the immobilized lipase for five cycles. This work provides a strategy for the preparation of an efficient biocatalyst for the synthesis of biodiesel.

The hydrophobicity of a support plays a critical role in the catalytic efficiency of immobilized lipases. 3-aminopropyltriethoxysilane (APTES)-modified silica clay (A-SC) was coupled with silane coupling agents of different alkyl chains (methyl triethoxysilane, vinyl triethoxysilane, octyl triethoxysilane, and dodecyl triethoxysilane) to prepare a series of hydrophobic support for lipase immobilization. The lipases were immobilized onto the support by conducting glutaraldehyde cross-linking processes. The results showed that the activity of the immobilized biocatalyst increased with hydrophobicity. The hydrolytic activity of Lip-Glu-C12-SC (contact angle 119.8°) can reach 5900 U/g, which was about three times that of Lip-Glu-A-SC (contact angle 46.5°). The immobilized lipase was applied as a biocatalyst for biodiesel production. The results showed that the catalytic yield of biodiesel with highly hydrophobic Lip-Glu-C12-SC could be as high as 96%, which is about 30% higher than that of Lip-Glu-A-SC. After being recycled five times, the immobilized lipase still maintained good catalytic activity and stability. This study provides a good strategy to improve the efficiency of immobilized lipases, showing great potential for future industrial application on biodiesel production.

In the extraction of collagens from mammalian tissues, the free pepsin used in the acid–pepsin extraction system is hard to recycle, and there is a risk of enzyme protein contamination in the extracted collagen products, which limits their applications.