CALB Immobilized onto Magnetic Nanoparticles for Efficient Kinetic Resolution of Racemic Secondary Alcohols: Long-Term Stability and Reusability

• Published in: Molecules
• Main Authors: Xiu Xing, Jun-Qi Jia, Jing-Fan Zhang, Zi-Wen Zhou, Jun Li, Na Wang, Xiao-Qi Yu
• Format: Article in Journal/Newspaper
• Language: English
• Published: MDPI AG 2019
• Subjects: CALB; magnetic nanoparticles; cross-linked enzyme aggregates; resolution; secondary alcohols; Organic chemistry; QD241-441; Antarc*; Antarctica
• Online Access: https://doi.org/10.3390/molecules24030490; https://doaj.org/article/45b573a0dad3473983b77ea541883bd1

In this study, an immobilization strategy for magnetic cross-linking enzyme aggregates of lipase B from Candida antarctica (CALB) was developed and investigated. Magnetic particles were prepared by conventional co-precipitation. The magnetic nanoparticles were modified with 3-aminopropyltriethoxysilane (APTES) to obtain surface amino-functionalized magnetic nanoparticles (APTES⁻Fe 3 O 4 ) as immobilization materials. Glutaraldehyde was used as a crosslinker to covalently bind CALB to APTES⁻Fe 3 O 4 . The optimal conditions of immobilization of lipase and resolution of racemic 1-phenylethanol were investigated. Under optimal conditions, esters could be obtained with conversion of 50%, enantiomeric excess of product (ee p ) > 99%, enantiomeric excess of substrate (ee s ) > 99%, and enantiomeric ratio (E) > 1000. The magnetic CALB CLEAs were successfully used for enzymatic kinetic resolution of fifteen secondary alcohols. Compared with Novozym 435, the magnetic CALB CLEAs exhibited a better enantioselectivity for most substrates. The conversion was still greater than 49% after the magnetic CALB CLEAs had been reused 10 times in a 48 h reaction cycle; both ee s and ee p were close to 99%. Furthermore, there was little decrease in catalytic activity and enantioselectivity after being stored at −20 °C for 90 days.