Immobilization of Enzymes via Microcontact Printing and Thiol–Ene Click Chemistry

Abstract

This Communication describes a bioconjugation method for the generation of enzyme microarrays on surfaces using photochemical thiol-ene chemistry in combination with microcontact printing. Glucose oxidase and lactase were readily immobilized (i.e., printing time 2 min) on alkene terminated self-assembled monolayers on glass as demonstrated by X-ray photoelectron spectroscopy and fluorescence microscopy. Furthermore, the activity of both immobilized enzymes was confirmed in single enzyme as well as cascade transformations.

Surface Functionalization with Carboxylic Acids by Photochemical Microcontact Printing and Tetrazole Chemistry

“…7 By this method, biotin and galactoside microarrays 4,8 could be fabricated as well as multifunctional surfaces 9 and enzyme microarrays. 10 Along with the use of photomasks, microcontact printing can be a method of choice to prepare patterned surfaces in (sub-) micrometer-sized resolutions. In microcontact printing, a patterned elastomer stamp is used to create surface functionalization.…”

Thiol–ene photoimmobilization of chymotrypsin on polysiloxane gels for enzymatic peptide synthesis

“…To this end, the light-stimulated thiol-ene [31][32][33] photoimmobilization, which utilizes multifunctional mercapto and vinyl monomers to form polymer networks, provides an alternative approach as a mild, efficient and green method for surface immobilization purposes. [34][35][36] Herein, we report a facile and fast thiol-ene photoimmobilization of olen-terminated enzyme onto poly[3mercaptopropylmethylsiloxane] (PMMS). [37][38][39][40] Chymotrypsin (CT) is chosen as a modal biocatalyst for peptide synthesis in this study because it possesses many accessible amino groups of side-chain-exposed amino acids (such as Lys, Ser, Gly, Cys, and Tyr residues) which can be chemically modied.…”

One-step purification and oriented attachment of protein A on silica and graphene oxide nanoparticles using sortase-mediated immobilization

“…23 Diels-Alder cycloaddition, thiol-ene additions and oxime reaction are the other approaches that are frequently used for the oriented immobilization of proteins on properly activated supports. 24,25 Chemoenzymatic approach for covalent immobilization is another technique that can be used for site specic conjugation of proteins on solid supports. This methodology is based on utilization of the inherent chemoselectivity of enzymes.…”