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

Abstract

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–Fe3O4) as immobilization materials. Glutaraldehyde was used as a crosslinker to covalently bind CALB to APTES–Fe3O4. 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 (eep) > 99%, enantiomeric excess of substrate (ees) > 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 ees and eep were close to 99%. Furthermore, there was little decrease in catalytic activity and enantioselectivity after being stored at −20 °C for 90 days.

Enhancing the Thermo-Stability and Anti-Biofilm Activity of Alginate Lyase by Immobilization on Low Molecular Weight Chitosan Nanoparticles

“…Reusability can be defined as multi-use of the enzyme with the activity does not change much and it shown to be an important index for immobilized enzyme [36]. The main advantage of immobilized strategy is that it can increase the reusability of the enzyme preparation.…”

Microbial Proteases Applications

“…Immobilization technology has been employed to obtain a high yield of alkaline proteases against a solid support of matrix (Kalisz, 1988; Rao et al, 1998). The proteases are usually immobilized in the alginate–chitosan beads, which exhibit reasonable stability and good activity at 47°C (Mehde et al, 2018; Xu et al, 2018; Özacar et al, 2019; Xing et al, 2019). Genetic engineering with the aim of hyperproduction of enzyme, cost-effectiveness, and quality helps scientists to capture the biotechnology market worldwide.…”

“Fishing and Hunting”—Selective Immobilization of a Recombinant Phenylalanine Ammonia-Lyase from Fermentation Media

“…The advantage of the last two methods is that the undesirable enantiomer of a racemate can be utilized via a racemization step or it is not even forming in the biotransformation. During the preceding decades, several valuable biotransformations have been implemented: resolution processes via hydroxyl or amine functional groups [15,16,17,18,19,20,21,22], oxidations [23,24,25] and reductions [26,27,28,29] in one step or in cascade reactions [30,31,32], production of amines with transaminases [30,32,33,34], imine reductases [28,35] or amine dehydrogenase [34,36] and amino acid formations [37,38,39]. Most of these reactions have industrial importance not only in pharmaceutical field [3,40] but in food [41,42,43,44,45] and fine chemical industries [46,47,48] or in modern analytics and diagnostic technologies as well [43,49,50,51,52,53].…”