Candida rugosa lipase nanoparticles as robust catalyst for biodiesel production in organic solvents

Inexpensive but resourceful sources of lipids, for example, used cooking oil (UCO) and brown grease (BG), which often contain large amounts of free fatty acids (FFA), are difficult to convert into biodiesel economically and in good yield. Candida rugosa lipase nanoparticles (cNP) were formed first and subsequently cross-linked nanoparticles (CLNP) were obtained by crosslinking of them. Alternatively, cNP were conjugated to magnetic nanoparticles (mNP) to achieve a cNP-mNP conjugate. All three formulations were employed in three different organic solvents (n-heptane, 1,4-dioxane, and t-butanol) to produce biodiesel using BG and UCO in the transesterification reaction with ethanol and methanol. The radii of nanoparticles (NP) were 5.5, 75, 100, 85 nm for mNP, cNP, CLNP, and cNP-mNP, respectively, as measured by scanning/transmission electron microscopy and dynamic light scattering. The catalytic efficiency (Kcat/KM) of cNP, CLNP, and cNP-mNP was increased ca. -25, -68, -176 folds in n-heptane and -35, -131, -262 folds in 1,4-dioxane compared to the lyophilized lipase in the model transesterification reaction of p-nitrophenyl palmitate (PNPP) with ethanol. In biodiesel formation, the best performance with 100% conversion of BG was achieved under optimum conditions with cNP-mNP, ethanol at a 1:3 molar ratio of lipid-to-alcohol, NP at a 1:0.1 weight ratio of lipid-to-enzyme, and water at a 1:0.04 weight ratio of enzyme-to-water at 30 oC for 35 h. The operational stability of the CLNP and cNP-mNP was sustained even after five consequent biodiesel batch conversions while 50% and 82% residual activity (storage stability) were retained after 40 d.