Dual-stimuli-responsive porous polymer enzyme reactor for tuning enzymolysis efficiency

A strategy for preparing a dual-stimuli-responsive porous polymer membrane enzyme reactor (D-PPMER) is described, consisting of poly (styrene-maleic anhydride-N-isopropylacrylamide-acrylate-3',3'-dimethyl-6-nitro-spiro[2H-1-benzopyran-2,2'-indoline]-1'-esterspiropyran ester) [P(S-M-N-SP)] and D-amino acid oxidase. Tunable control via "on/off" 365 nm UV light irradiation and temperature variation was used to change the membrane surface configuration and adjust the enzymolysis efficiency of the D-PPMER. A chiral capillary electrophoresis technique was developed for evaluation of the enzymatic efficiency of D-PPMER with a Zn(II)-dipeptide complex as the chiral selector and D,L-serine as the substrate. Interestingly, the enzymatic kinetic reaction rate of D-PPMER under UV irradiation at 36 degrees C (9.2 x -10(-2) mM.min(-1)) was 3.2-fold greater than that of the free enzyme (2.9 x -10(-2) mM.min(-1)). This was because upon UV irradiation at high temperature, the P(SP) and P(N) moieties altered from a "stretched" to a "curled" state to encapsulate the enzyme in smaller cavities. The confinement effect of the cavities further improved the enzymatic efficiency of the D-PPMER. This protocol highlights the outstanding potential of smart polymers, enables tunable control over the kinetic rates of stimuli-responsive enzyme reactors, and establishes a platform for adjusting enzymolysis efficiency using two different stimuli.