Electric‐field‐enhanced selective separation of products of an enzymatic reaction in a membrane micro‐contactor

• Published in: Biotechnology and bioengineering 2021-02, Vol.118 (2), p.715-724
• Main Authors: Romanov, Alexandr, Slouka, Zdeněk, Přibyl, Michal
• Format: Article
• Language: English
• Subjects: 6‐aminopenicillanic acid; Aminopenicillanic acid; Antibiotics; Dialysis; Diffusion; Direct current; electric field; Electric fields; Enzymes; Mass transfer; Membranes; microextraction; Microfluidics; Penicillin; Penicillin acylase; Phenylacetic acid; Reaction products; Residence time distribution; Selectivity; Semipermeable membranes; Separation
• ISSN: 0006-3592; 1097-0290
• DOI: 10.1002/bit.27597

Processes employed in separations of products of enzyme reactions are often driven by diffusion, and their efficiency can be limited. Here, we exploit the effect of a direct current (DC) electric field that intensifies mass transfer through a semipermeable membrane for fast, continuous, and selective separation of electrically charged molecules. Specifically, we separate low‐molecular‐weight reaction products (phenylacetic acid, 6‐aminopenicillanic acid) from the original reaction mixture containing a free enzyme (penicillin acylase). The developed microfluidic dialysis‐membrane contactor allows a stable counter‐current arrangement of the retentate and permeates liquid streams on which DC electric field is perpendicularly applied. The applied electric field significantly accelerates the transport of electrically charged products through the semipermeable membrane yielding high separation efficiencies at short residence times. The residence time of 5 min is sufficient to reach 100% separation yield in the electric field. The same residence time provides only a 50% yield in the diffusion‐controlled experiments. We experimentally demonstrated that a combined microreactor–microextractor with a recycle of the soluble penicillin acylase can continuously produce both the reaction products at high concentrations. The developed membrane‐contactor is a versatile platform allowing to tune its characteristics, such as selectivity given by the membrane, or the type of the retentate phase, for a specific application. Direct current (DC) electric field intensified mass transfer through a semipermeable membrane for fast, continuous, and selective separation of phenylacetic acid and 6‐aminopenicillanic acid from the original reaction mixture containing a free penicillin acylase. The residence time of 5 min is sufficient to reach 100% separation yield. The developed microfluidic dialysis‐membrane contactor allows a stable counter‐current arrangement of the retentate and permeates liquid streams on which DC electric field is perpendicularly applied.