Lipase B from Candida antarctica Immobilized on Epoxy-functionalized Hollow Silica Microspheres: Efficient Biocatalysts for Enantiomer Selective Acylation of Alcohols and Amines

Abstract

Hollow silica microspheres with promising physical properties (MAT540TM) as support for enzyme immobilization and biocatalyst were investigated in this study. The amine-functionalized MAT540TM was activated by six bisepoxides inclosing different spacers and used as epoxy-functionalized carrier for immobilization of lipase B from Candida antarctica (CaLB). The novel, covalently fixed CaLB biocatalysts were compared in kinetic resolution (KR) of racemic 1-phenyethanol rac-1 and five racemic amines rac-3a-e using shaken flasks and continuous-flow packed-bed microreactors. Mechanic stability, re-usability and the effect of temperature (0–90 °C) on productivity and enantiomer selectivity of the covalently immobilized CaLB were investigated. The best performing CaLB biocatalyst showed good mechanic stability after 24 h operation time in continuous-flow mode at 60 °C and provided in KRs of racemic 1-phenyethanol rac-1 with vinyl acetate and of five racemic amines with isopropyl 2-ethoxyacetate as acylating agent the non-reacted (S)-alcohol [(S)-1] or (S)-amines [(S)-3a-e] and the forming (R)-ester [(R)-2] or (R)-amide [(R)-4a-e] in good yields with high enantiomeric excess (ee > 99 %, for all).

Conservation of the Biocatalytic Activity of Whole Yeast Cells by Supported Sol – Gel Entrapment for Efficient Acyloin Condensation

“…In case of the 2 nd generation sol -gel method, wholecells and a proper inert solid support were co-immobilized. Hollow silica microspheres with amino/vinyl grafted surface (M540) were previously described as efficient enzyme carrier [41,42], thus M540 was tried in 2 nd generation sol -gel systems as support material as well.…”

Advances in Recombinant Lipases: Production, Engineering, Immobilization and Application in the Pharmaceutical Industry

“…The carrier hollow silica microspheres (MAT540TM) activated by six bisepoxides enclosing different spacers was applied for the immobilization of CALB. The immobilized preparation was used in the kinetic resolution of racemic 1-phenyethanol and five racemic amines using shaken flasks and continuous-flow packed-bed microreactors with good yields and high enantiomeric excess (ee > 99%, for all) [144]. Furthermore, the recombinant CALB expressed in K. phaffii was immobilized by cross-linked enzyme aggregate (CLEA) technology and tested for the synthesis of olvanil [145].…”

“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].…”

“…Abundantly, glutaraldehyde is applied for the formation of covalent bonds [105]. Besides the often used glutaraldehyde, epoxy functioned supports have been used for a long time [106], and activated carboxyl, amino, hydroxyl or thiol groups are also suitable for covalent immobilization [21,104,107,108,109]. The two-step method of the covalent immobilization can be enhanced with heterofunctional epoxy supports [109,110,111,112].…”

“…Also the pH during the immobilization has a great impact on the formation of stabile multipoint covalent attachment between the enzyme and the support [114,120]. Nowadays, a great variety of supports are used: macroporous polymers [107,109], silica particles in different size and shape [21], magnetic nanoparticles [121] and several type of nanofibers [122] or nanotubes [123] are just some of the most popular ones. Also modification of the surface with inert groups besides the reactive functions has an effect on the biocatalytic activity [103,109].…”