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

ABSTRACT: Chemical incorporation of enzymes onto polymeric materials has recently attracted intense scientific attention. Cross-linked polysiloxane gels as a typical super-hydrophobic support, are a good candidate for supporting enzymes in low-water organic medium to efficiently catalyze peptide synthesis because the hydrophobic polysiloxane matrix can prevent water from attacking the acyl-enzyme intermediate, which is beneficial for the shift in equilibrium to peptide formation. In this work, we develop a facile strategy to photoimmobilize olefin-functionalized chymotrypsin onto cross-linked polysiloxane gels via UV-initiated thiol-ene click chemistry. The impacts of water addition amount, heat-treatment and recyclability of the immobilized chymotrypsin influencing the peptide synthesis efficiency are investigated. Compared with the native chymotrypsin, polysiloxane-immobilized chymotrypsin showed advantageous catalytic activity, higher thermal stability and superior recyclability.

SUBMITTER: Wang M

PROVIDER: S-EPMC9079220 | biostudies-literature | 2018 Mar

REPOSITORIES: biostudies-literature

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

Wang Meng M Xing Jun J Sun Yu-Tang YT Guo Ling-Xiang LX Lin Bao-Ping BP Yang Hong H

RSC advances 20180327 22

Chemical incorporation of enzymes onto polymeric materials has recently attracted intense scientific attention. Cross-linked polysiloxane gels as a typical super-hydrophobic support, are a good candidate for supporting enzymes in low-water organic medium to efficiently catalyze peptide synthesis because the hydrophobic polysiloxane matrix can prevent water from attacking the acyl-enzyme intermediate, which is beneficial for the shift in equilibrium to peptide formation. In this work, we develop ...[more]

PMID: 35539381

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Project description:Vascular endothelial growth factor (VEGF) plays a vital role in promoting attachment and proliferation of endothelial cells, and induces angiogenesis. In recent years, much research has been conducted on functionalization of tissue engineering scaffolds with VEGF or VEGF-mimetic peptide to promote angiogenesis. However, most chemical reactions are nonspecific and require organic solvents, which can compromise control over functionalization and alter peptide/protein activity. An attractive alternative is the fabrication of functionalizable electrospun fibers, which can overcome these hurdles. In this study, we used thiol-ene chemistry for the conjugation of a VEGF-mimetic peptide to the surface of poly (ε-caprolactone) (PCL) fibrous scaffolds with varying amounts of a functional PCL-diacrylate (PCL-DA) polymer. 30% PCL-DA was selected due to homogeneous fiber morphology. A VEGF-mimetic peptide was then immobilized on PCL-DA fibrous scaffolds by a light-initiated thiol-ene reaction. 7-Mercapto-4-methylcoumarin, RGD-FITC peptide and VEGF-TAMRA mimetic peptide were used to validate the thiol-ene reaction with fibrous scaffolds. Tensile strength and elastic modulus of 30% PCL-DA fibrous scaffolds were significantly increased after the reaction. Conjugation of 30% PCL-DA fibrous scaffolds with VEGF peptide increased the surface water wettability of the scaffolds. Patterned structures could be obtained after using a photomask on the fibrous film. Moreover, in vitro studies indicated that scaffolds functionalized with the VEGF-mimetic peptide were able to induce phosphorylation of VEGF receptor and enhanced HUVECs survival, proliferation and adhesion. A chick chorioallantoic membrane (CAM) assay further indicated that the VEGF peptide functionalized scaffolds are able to promote angiogenesis in vivo. These results show that scaffold functionalization can be controlled via a simple polymer mixing approach, and that the functionalized VEGF peptide-scaffolds have potential for vascular tissue regeneration.

Dataset Information

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

Publications

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

Similar Datasets

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