Simple Technique for Preparing Stable and Recyclable Cross-Linked Enzyme Aggregates with Crude-Pored Microspherical Silica Core

Abstract

With the aim of providing a highly stable and easy recyclable biocatalyst, we prepared a new biocatalyst formulation based on cross-linked enzyme aggregate (CLEA) technology by the physical adsorption of cross-linked phenylalanine ammonia lyase on a highly porous microspherical silica gel solid support (S-CLEAs). The main factors in the preparation of S-CLEAs were optimized by response surface methodology (RSM). Analysis by scanning electron microscopy (SEM), FT-IR spectroscopy, and confocal laser scanning microscopy (CLSM) indicated that the CLEAs were successfully immobilized on the crude-pored microspherical silica gel. Compared to standard CLEAs, the S-CLEAs can be easily separated from the reaction mixture by natural precipitation without centrifugation and filtration treatments. Although the S-CLEAs revealed an excellent stability of the enzyme against high temperature and denaturants, the volumetric activity of the S-CLEAs decreased because of the presence of silica gel. In a word, the S-CLEA technology in this study might open an attractive route toward effectively improving the catalytic properties of traditional CLEAs.

Encapsulation of Spherical Cross-Linked Phenylalanine Ammonia Lyase Aggregates in Mesoporous Biosilica

“…Therefore, it is important to control the particle size of CLEAs and to improve their mechanical stability. , To overcome these problems, CLEAs of papain in commercial macroporous silica gel were prepared and showed excellent operation stability . The formation of CLEAs of phenylalanine ammonia lyase (PAL) from Rhodotorula glutinis into a macroporous silica gel exhibited good reusability . The magnetic CLEAs of α-amylase were prepared on amino-functionalized magnetite nanoparticles and exhibited excellent thermostability, storage stability, and reusability .…”

Bio-based and cost effective method for phenolic compounds removal using cross-linked enzyme aggregates

“…In addition, the combination of traditional techniques with CLEAs technology may prove useful, as in the case of the adsorbed cross-linked phenylalanine ammonia lyase aggregate on the crude-pored microspherical silica core. The immobilized CLEAs can be precipitated naturally without filtration or centrifugation, thus reducing mass-transfer limitations (Cui et al, 2014). However, it is important to take into consideration the biochemical properties and protein structure of each enzyme before proceeding to any immobilization (Yamaguchi et al, 2018).…”

Immobilization of Phenylalanine Ammonia-lyase via EDTA Based Metal Chelate Complexes – Optimization and Prospects

“…Thus, phenylalanine ammonia-lyase from Petroselinum crispum (parsley) (PcPAL) having a tetrameric structure and promising biocatalytic activity has been selected as target enzyme for this study. So far mostly covalent immobilization methods [47,48,[53][54][55][56][57] and encapsulation techniques [58,59] have been studied for PAL enzymes, another affinity-based method have already been described in our previous work [24].…”