A Continuous Bioreactor Prepared via the Immobilization of Trypsin on Aldehyde-Functionalized, Ring-Opening Metathesis Polymerization-Derived Monoliths

作者：Sudheendran Mavila; Buchmeiser Michael R^*

来源：Macromolecules, 2010, 43(23): 9601-9607.

DOI：10.1021/ma101922s

The ring-opening metathesis polymerization (ROMP) of norborn-2-ene (NBE) and cis-cyclooctene (COE) was initiated with well-defined Grubbs-type initiators, i.e., RuCl(2)(CHPh)(PCy(3))(2) (1), [RuCl(2)(PCy(3))(IMesH(2))(CHPh)] (2), and [RuCl(2)(3-Br-Py)(2)(IMesH(2))(CHPh)] (3) (MesH(2) = 1,3-bis(2,4,6-trimethylphenyl)imidazolin-2-ylidene, PCy(3) = tricyclohexylphosphine, 3-Br-Py = 3-bromopyridine). Reaction of the living polymers with O(2) (air) resulted in the formation of aldehyde-semitelechelic polymers in up to 80% yield, depending on the initiator and monomer used. To proof aldehyde formation, the terminal aldehyde groups were converted into the corresponding 2,4-dinitrophenylhydrazine derivatives, and the structure of the hydrazones was confirmed by NMR and IR spectroscopy. This simple methodology was then used for the functionalization of ROMP-derived monoliths prepared from NBE, 1,4,4a,5,8,8a-hexahydro-1,4,5,8-exo-endo-dimethanonaphthalene (DMN-H6) and (NBE-CH(2)O)(3)SiCH(3), to yield aldehyde-funtionalized monoliths. The extent of aldehyde formation was determined by hydrazone formation. Up to 8 mu mol of aldehyde groups/g monolith could be generated by this approach. Finally, these aldehyde-functionalized monoliths were used for the immobilization of trypsin. Excellent proteolytic activity of the immobilized enzyme was found both under batch and continuous flow conditions.

• 出版日期2010-12-14