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Synergistic Enhancement of Enzyme Performance and Resilience via Orthogonal Peptide–Protein Chemistry Enabled Multilayer Construction
Biomacromolecules ( IF 5.5 ) Pub Date : 2018-05-16 00:00:00 , DOI: 10.1021/acs.biomac.8b00306
Xue-Jian Zhang 1, 2 , Xiao-Wei Wang 2 , Jia-Xing Sun 1 , Chao Su 1 , Shuguang Yang 1 , Wen-Bin Zhang 2
Affiliation  

Protein immobilization is critical to utilize their unique functions in diverse applications. Herein, we report that orthogonal peptide–protein chemistry enabled multilayer construction can facilitate the incorporation of various folded structural domains, including calmodulin in different states, affibody, and dihydrofolate reductase (DHFR). An extended conformation is found to be the most advantageous for steady film growth. The resulting protein thin films exhibit sensitive and selective responsive behaviors to biosignals, such as Ca2+, trifluoperazine, and nicotinamide adenine dinucleotide phosphate (NADPH), and fully maintain the catalytic activity of DHFR. The approach is applicable to different substrates such as hydrophobic gold and hydrophilic silica microparticles. The DHFR enzyme can be immobilized onto silica microparticles with tunable amounts. The multilayer setup exhibits a synergistic enhancement of DHFR activity with increasing numbers of bilayers and also makes the embedded DHFR more resilient to lyophilization. Therefore, this is a convenient and versatile method for protein immobilization with potential benefits of synergistic enhancement in enzyme performance and resilience.

中文翻译:

通过正交肽-蛋白质化学实现多层构建,协同增强酶的性能和弹性

蛋白质固定化对于在各种应用中发挥其独特功能至关重要。在本文中,我们报道了正交肽-蛋白质化学能够实现多层构建,可以促进各种折叠结构域的整合,包括不同状态的钙调蛋白,亲和体和二氢叶酸还原酶(DHFR)。发现延长的构象对于稳定的膜生长是最有利的。所得的蛋白质薄膜对生物信号(例如Ca 2+)表现出敏感和选择性的响应行为,三氟拉嗪和烟酰胺腺嘌呤二核苷酸磷酸(NADPH),并完全保持DHFR的催化活性。该方法适用于不同的基底,例如疏水性金和亲水性二氧化硅微粒。DHFR酶可以固定在二氧化硅微粒上。多层装置显示出随着双层数的增加,DHFR活性协同增强,并且还使嵌入的DHFR对冻干更具弹性。因此,这是一种方便且通用的蛋白质固定方法,具有协同增强酶性能和弹性的潜在好处。
更新日期:2018-05-16
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