Rapid Identification of an Extraction System for the Purification of Enzymes

Industrial relevant enzymes have reached industrial competitiveness over classical chemical catalysts due to gentle
operating conditions and high specificity for the substrate. Within their production, the purification of the enzyme using
Aqueous Two-Phase Extraction (ATPE) has been shown to be a promising alternative to time- and cost-intensive
chromatographic separation. Within this work, we applied an optimized method to identify tailor-made Aqueous Two-Phase
Systems (ATPS) to be used in ATPE. It was shown, that for an industrially relevant enzyme (that is Chimera [supplied by
Georgia Tech, Bommarius Lab]) it was possible to identify a suitable ATPS as well as a process window for the ATPE in
less than three weeks. With achievable yields above 78%, the possibility to influence the partitioning behavior of Chimera,
this system outclasses the previously randomly selected ATPS (Trial-and-Error High Throughput Screening) by a factor of
four, at the same time keeping precipitation of the enzyme below 17% (previously 70%).