A versatile ceramic capillary membrane reactor system for continuous enzyme‐catalyzed hydrolysis

Abstract

As an alternative to classical batch processes, enzyme-catalyzed hydrolysis can also be carried out continuously. To facilitate this, a continuous ceramic capillary membrane reactor system (CCCMRS) was developed which can be operated with various proteolytic enzymes immobilized on the porous ceramic capillary membranes. This system has several advantages over common batch processes regarding stability, reproducibility and controllability and can easily be adapted to optimal reaction conditions and individual preferences. Two exemplary applications utilizing the CCCMRS were carried out and investigated in long-term stability studies. In the first application the continuous enzymatic cleavage of human IgG into the antibody fragments Fab and Fc by immobilized papain was performed. A total volume of 22 mL of 1 mg mL -1 IgG-solution was enzymatically cleaved over a period of 33.3 h. The antibody cleavage products could be detected in an SEC-HPLC over the whole process time thus indicating long-term stability of the continuous hydrolysis process. The second application investigated the continuous digestion of pea and almond protein isolates by immobilized Alcalase resulting in the generation of a large variety of different peptides. This peptide fingerprint remains constant over a long period of time enabling fractionation and thus making the peptides accessible for further bioactivity studies in sufficient quantities. The constant peptide fingerprint could be shown in the RP-HPLC analysis for all 30 samples with a total volume of 29.7 mL collected over a period of 45 h.

Microfluidic Immobilized Enzymatic Reactors for Proteomic Analyses—Recent Developments and Trends (2017–2021)

“…Furthermore, µ-IMERs are often prepared in a relatively low-flow format, such as a capillary. However, if the inner diameter of these capillaries reaches 1 mm (e.g., [85][86][87]) then these devices cannot be considered microfluidic systems. µ-IMERs are generally very small devices/chips consisting of channels/capillaries with inner diameters less than a few hundred micrometers.…”

“…Enzymes have also been attached to porous ceramic capillary membranes [85,86]. The tubular shape with an inner diameter of ~1 mm was prepared by extrusion.…”

Advanced enzymatic hydrolysis of food proteins for the production of bioactive peptides

“…Proteolytic enzymes have been assayed on different types of support materials. For instance, ceramic capillary membranes were used for immobilising Alcalase and its suitability was proved with the hydrolysis of casein, sunflower, lupin, pea and almond proteins [53]. Hollow mesoporous silica was modified with a variety of metal ions chelated through triamino ligands.…”

Liposome–Papain Conjugates for Catalytic Digestion of Antibody Producing Fab Fragments

“…Immobilized papain is, therefore, useful to develop economic biocatalytic processes with continuous or recycling use of the catalyst, giving catalyst-free products. Papain was applied in its various immobilized forms as a catalyst for producing peptides from food proteins, processing milk to remove allergenic proteins, an electrochemical reaction, and synthesizing peptides. , Carrier materials used for the papain immobilization were elaborated in terms of their structure, interfacial property, , or solution condition-dependent solubility, so that the immobilized papain could exhibit catalytic activity toward macromolecular substrates and the catalyst could be recovered for reuse. Furthermore, the physicochemical properties of carrier materials are also important for the immobilized papain to keep its biologically active conformation and proper molecular orientation on a solid surface. , …”