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Facile Preparation of an Enzyme‐Immobilized Microreactor using a Cross‐Linking Enzyme Membrane on a Microchannel Surface
Author(s) -
Honda Takeshi,
Miyazaki Masaya,
Nakamura Hiroyuki,
Maeda Hideaki
Publication year - 2006
Publication title -
advanced synthesis and catalysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.541
H-Index - 155
eISSN - 1615-4169
pISSN - 1615-4150
DOI - 10.1002/adsc.200606224
Subject(s) - microreactor , chemistry , immobilized enzyme , microchannel , polymerization , biocatalysis , laminar flow , microfluidics , chromatography , enzyme , chemical engineering , polymer , organic chemistry , nanotechnology , catalysis , materials science , reaction mechanism , physics , engineering , thermodynamics
The enzyme microreactor has considerable potential for use in biotechnological syntheses and analytical studies. Simplifying the procedure of enzyme immobilization in a microreactor is attractive, and it is achievable by utilizing enzyme immobilization techniques and taking advantage of the characteristics of microfluidics. We previously developed a facile and inexpensive preparation method for an enzyme‐immobilized microreactor. The immobilization of enzymes can be achieved by the formation of an enzyme‐polymeric membrane on the inner wall of the microchannel through cross‐linking polymerization in a laminar flow. However, this method is unsuitable for use in conjunction with electronegative enzymes. Therefore, a novel preparation method using poly‐ L ‐lysine [poly(Lys)] as a booster and an adjunct for the effective polymerization of electronegative enzymes was developed in this study. Using aminoacylase as a model for an electronegative enzyme, the reaction conditions for the enzyme‐cross‐linked aggregation were optimized. On the basis of the determined conditions, an acylase‐immobilized tubing microreactor was successfully prepared by cross‐linking polymerization in a concentric laminar flow. The resulting microreactor showed a higher stability against heat and organic solvents compared to those of the free enzyme. The developed method using poly(Lys) was applicable to various enzymes with low isoelectric points, suggesting that this microreactor preparation utilizing a cross‐linked enzyme in a laminar flow could be expanded to microreactors in which a broad range of functional proteins are employed.