Inversion of Enzymatic Enantiocatalysis Mediated by Biomolecular Condensates.

Herein, we report the enantioselective phosphodiesterase behavior of an enzyme, alkaline phosphatase (ALP) (using both R- and S-forms of the RNA-model substrate, 2-hydroxypropyl-p-nitrophenyl phosphate (HPNPP)), in protein (bovine serum albumin (BSA)) and DNA-based biocondensates. The native preference toward ALP enzyme is toward S-HPNPP in aqueous buffer with an enantiomeric excess ratio (E) value of around 33%. This preference for S-HPNPP remains intact when ALP is encapsulated in BSA condensate (E-value range: 10 to 34%). A complete reversal of the E value (-20 to -47%) is observed in the DNA-based complex coacervate, as ALP here reacts faster with R-HPNPP. We finally demonstrate that the reversal of enzyme enantioselectivity in the condensed phase is linked to the enzyme's conformational dynamics inside the coacervate. This work will be important in synthetic protocells studies, as it provides novel insights into enzyme promiscuity and chiral selectivity in the diversified internal environment of biocondensates.