Citation: Juan Qiao, Lili Liu, Ji Shen, Li Qi. Enzyme immobilization on a pH-responsive porous polymer membrane for enzymatic kinetics study[J]. Chinese Chemical Letters, ;2021, 32(10): 3195-3198. doi: 10.1016/j.cclet.2021.03.021 shu

Enzyme immobilization on a pH-responsive porous polymer membrane for enzymatic kinetics study

Juan Qiao ^{a, b} ,
Lili Liu ^{a, c} ,
Ji Shen ^{a, b} ,
Li Qi ^{a, b, *,}

a.
Beijing National Laboratory of Molecular Sciences, Key Laboratory of Analytical Chemistry for Living Bio-systems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China
b.
School of Chemical Sciences, University of Chinese Academy of Sciences, Beijing 100049, China
c.
College of Chemistry & Environmental Science, Hebei University, Baoding 071002, China

qili@iccas.ac.cn

Received Date: 9 December 2020
Revised Date: 20 January 2021
Accepted Date: 9 March 2021
Available Online: 11 March 2021

Figures(4)

Immobilization of enzymes onto carriers is a rapidly growing research area aimed at increasing the stability, reusability and enzymolysis efficiency of free enzymes. In this work, the role of phase-separation and a pH-responsive "hairy" brush, which greatly affected the topography of porous polymer membrane enzyme reactors (PMER), was explored. The porous polymer membrane was fabricated by phase-separation of poly(styrene- co-maleic anhydride-acrylic acid) and poly(styrene-ethylene glycol). Notably, the topography and pores size of the PMER could be controlled by phase-separation and a pH-responsive "hairy" brush. For evaluating the enzymolysis efficiency of d-amino acid oxidase (DAAO) immobilized carrier (DAAO@PMER), a chiral ligand exchange capillary electrophoresis method was developed with d-methionine as the substrate. The DAAO@PMER showed good reusability and stability after five continuous runs. Notably, comparing with free DAAO in solution, the DAAO@PMER exhibited a 17.7-folds increase in catalytic velocity, which was attributed to its tailorable topography and pH-responsive property. The poly(acrylic acid) moiety of poly(styrene- co-maleic anhydride-acrylic acid) as the pH-responsive "hairy" brush generated topography changing domains upon adjusting the buffer pH, which enable the enzymolysis efficiency of DAAO@PMER to be tuned based upon the well-deﬁned architectures of the PMER. This approach demonstrated that the topographical changes formed by phase-separation and the pH-responsive "hairy" brush indeed made the proposed porous polymer membrane as suitable supports for enzyme immobilization and fitting for enzymolysis applications, achieving high catalytic performance.
- Enzyme immobilization,
- Porous polymer membrane topography,
- Phase separation,
- pH-stimuli responsive hairy brush
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