Construction of a thermoresponsive magnetic porous polymer membrane enzyme reactor for glutaminase kinetics study

Liping Zhao, Juan Qiao, Meyong Hee Moon, Li Qi

Research output: Contribution to journalArticle

Abstract

Fabrication of polymer membranes with nanopores and a confinement effect toward enzyme immobilization has been an enabling endeavor. In the work reported here, an enzyme reactor based on a thermoresponsive magnetic porous block copolymer membrane was designed and constructed. Reversible addition–fragmentation chain transfer polymerization was used to synthesize the block copolymer, poly(maleic anhydride–styrene–N-isopropylacrylamide), with poly(N-isopropylacrylamide) as the thermoresponsive moiety. The self-assembly property of the block copolymer was used for preparation of magnetic porous thin film matrices with iron oxide nanoparticles. By covalent bonding of glutaminase onto the surface of the membrane matrices and changing the temperature to tune the nanopore size, we observed enhanced enzymolysis efficiency due to the confinement effect. The apparent Michaelis–Menten constant and the maximum rate of the enzyme reactor were determined (Km = 32.3 mM, Vmax = 33.3 mM min−1) by a chiral ligand exchange capillary electrochromatography protocol with l-glutamine as the substrate. Compared with free glutaminase in solution, the proposed enzyme reactor exhibits higher enzymolysis efficiency, greater stability, and greater reusability. Furthermore, the enzyme reactor was applied for a glutaminase kinetics study. The tailored pore sizes and the thermoresponsive property of the block copolymer result in the designed porous membrane based enzyme reactor having great potential for high enzymolysis performance. [Figure not available: see fulltext.].

Original languageEnglish
Pages (from-to)5211-5218
Number of pages8
JournalAnalytical and Bioanalytical Chemistry
Volume410
Issue number21
DOIs
Publication statusPublished - 2018 Aug 1

Fingerprint

Glutaminase
Polymers
Block copolymers
Membranes
Kinetics
Enzymes
Nanopores
Enzyme immobilization
Capillary Electrochromatography
Reusability
Glutamine
Self assembly
Pore size
Polymerization
Immobilization
Nanoparticles
Ligands
Fabrication
Thin films
Substrates

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry

Cite this

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Construction of a thermoresponsive magnetic porous polymer membrane enzyme reactor for glutaminase kinetics study. / Zhao, Liping; Qiao, Juan; Moon, Meyong Hee; Qi, Li.

In: Analytical and Bioanalytical Chemistry, Vol. 410, No. 21, 01.08.2018, p. 5211-5218.

Research output: Contribution to journalArticle

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