Influence of Dlutaraldehyde Cross-Linking Modes on the Recyclability of Immobilized Lipase B from Candida antarctica for Transesterification of Soy Bean Oil

• Published in: Molecules (Basel, Switzerland) Vol. 23; no. 9; p. 2230
• Main Authors: A Modenez, Iago, Sastre, Diego E, C Moares, Fernando, Marques Netto, Caterina G C
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 02.09.2018
• Subjects: biodiesel synthesis; Biofuels; Candida - enzymology; Cross-Linking Reagents - chemistry; cross-linking types; enzyme immobilization; Enzymes, Immobilized - metabolism; Esterification; Glutaral - chemistry; Kinetics; lipase; Lipase - metabolism; magnetic nanoparticles; Magnetite Nanoparticles - chemistry; Models, Molecular; Soybean Oil - metabolism; Thermodynamics; enzyme immobilization; magnetic nanoparticles; biodiesel synthesis; lipase; cross-linking types
• ISSN: 1420-3049
• DOI: 10.3390/molecules23092230

Lipase B from (CAL-B) is largely employed as a biocatalyst for hydrolysis, esterification, and transesterification reactions. CAL-B is a good model enzyme to study factors affecting the enzymatic structure, activity and/or stability after an immobilization process. In this study, we analyzed the immobilization of CAL-B enzyme on different magnetic nanoparticles, synthesized by the coprecipitation method inside inverse micelles made of zwitterionic surfactants, with distinct carbon chain length: 4 (ImS4), 10 (ImS10) and 18 (ImS18) carbons. Magnetic nanoparticles ImS4 and ImS10 were shown to cross-link to CAL-B enzyme via a Michael-type addition, whereas particles with ImS18 were bond via pyridine formation after glutaraldehyde cross-coupling. Interestingly, the Michael-type cross-linking generated less stable immobilized CAL-B, revealing the influence of a cross-linking mode on the resulting biocatalyst behavior. Curiously, a direct correlation between nanoparticle agglomerate sizes and CAL-B enzyme reuse stability was observed. Moreover, free CAL-B enzyme was not able to catalyze transesterification due to the high methanol concentration; however, the immobilized CAL-B enzyme reached yields from 79.7 to 90% at the same conditions. In addition, the transesterification of lipids isolated from oleaginous yeasts achieved 89% yield, which confirmed the potential of immobilized CAL-B enzyme in microbial production of biodiesel.