The enzyme-microreactor has a strong potential for applications such as biotechnological syntheses and analyses. At present, interesting microreactors with various enzymes have been reported. However, the developed methods of microreactor preparations require high-level techniques and/or multi-step procedure which could lead to low cost performance. Also, disposability of the reactor is considered essential in bioanalyses. Simplifying the procedure of enzyme-immobilization in microreactor has been attractive. This is achievable by concertedly utilizing the reaction for enzyme-immobilization and the characteristic of microfluidics. We have designed an enzyme-immobilized microreactor possessing a substrate membrane that covers the internal surface of the micro-channel. The cylindrical membrane is composed of cross-linked polymerized enzyme product. This carrier-free method based on enzyme-cross-linked aggregation (CLEA) realized a facile and inexpensive preparation of an enzyme-immobilized microreactor. In this study, α-chymotrypsin was used as a model enzyme for protein digestion. Recently, microreactors capable of protein digestion has been attractive in proteomics studies. By utilizing the optimized condition in CLEA formation, α-chymotrypsin-immobilized tubing microreactor was successfully prepared by cross-linking polymerization in a concentric laminar flow. The resulting microreactor showed higher stability against heat and organic solvent than those of the free enzyme. These results show that this preparation method leads to a flexible technology platform for screening and designing a potential enzyme-microreactor for broad applications.
CITATION STYLE
Honda, T., Miyazaki, M., Nakamura, H., & Maeda, H. (2006). Immobilization of enzymes on microchannel surface through cross-linking polymerization. In Topical Conference on Applications of Micro-Reactor Engineering 2006, Held at the 2006 AIChE Spring National Meeting (pp. 103–110). AIChE.
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