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

机译:胰凝乳蛋白酶在聚硅氧烷凝胶上的巯基烯烃光固定化,用于酶促肽合成

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摘要

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.
机译:酶在聚合物材料上的化学掺入最近引起了强烈的科学关注。交联聚硅氧烷凝胶作为典型的超疏水载体,是在低水有机介质中支持酶以有效催化肽合成的良好候选者,因为疏水性聚硅氧烷基质可以防止水侵蚀酰基酶中间体,这是有益的平衡向肽形成的转变。在这项工作中,我们开发了一种简便的策略,可通过紫外线引发的硫醇-烯点击化学将烯烃官能化的胰凝乳蛋白酶光固定在交联的聚硅氧烷凝胶上。研究了固定化胰凝乳蛋白酶的加水量,热处理和可回收性对肽合成效率的影响。与天然胰凝乳蛋白酶相比,固定化聚硅氧烷的胰凝乳蛋白酶具有有利的催化活性,更高的热稳定性和可回收性。

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