Immobilized Dehydrogenases for the Biosynthesis of Phenylacetic Acids

Two phenylacetaldehyde dehydrogenases, originating from Escherichia coli K-12 (FeaB-K-12) and Sphingopyxis fribergensis Kp5.2 (FeaB-Kp5.2), were immobilized on powdery silica carrier with various functionalization. First, the suitability of these carriers for application in combination with phenylacetaldehydes and phenylacetic acids was studied. Out of two carriers functionalized differently, mesoporous cellular foam, whose surface was modified with 3-glycidyloxypropyl groups (MCF-G), showed promising results. Hence, this carrier was further tested at 17 different immobilization conditions. Despite both enzymes showing high immobilization efficiency, the initial activities were relatively low compared to the free enzymes. Interestingly, the immobilized FeaB-Kp5.2 on MCF-G-Kw showed about 80% of retained activity after two months of incubation at 0 °C, indicating that the immobilization enhances the stability of this enzyme. In contrast, no changes in the temperature stability of FeaB-Kp5.2 due to immobilization could be noted. However, relative enzyme activities towards all three substituted phenylacetaldehydes could be increased by the immobilization to approximately 130%. The most active and stable powdery immobilizate was MCF-G-Kw-FeaB-Kp5.2 at pH 8. In addition, FeaB-Kp5.2 was also immobilized and tested on monolith silica carrier for continuous catalysis to produce phenylacetic acids.