热稳定性
葡萄糖-6-磷酸异构酶
固定化酶
共价键
化学
肽
异构酶
酶
配体(生物化学)
亲和层析
组合化学
色谱法
生物化学
有机化学
受体
作者
Nian‐Qing Shi,Min Zheng,Xiaofeng Wu,Ning Chen,Luying Jiang,Baogen Chang,Fuping Lu,Fufeng Liu
标识
DOI:10.1021/acs.jafc.3c02644
摘要
A novel affinity peptide orientation and light-controlled covalent immobilized method was developed. Sucrose isomerase (SI) was selected as the model enzyme. Molecular simulation was first performed to select the targeted immobilization region. Subsequently, a short peptide (H2N-VNIGGX-COOH, VG) with high affinity to this region was rationally designed. Thereafter, 4-benzoyl-l-phenylalanine with the photosensitive group of benzophenone was introduced. Then, the affinity between the ligand and the SI was validated using molecular dynamics simulation. Thereafter, the SI was directionally immobilized onto the surface of the epoxy resin (EP) guided by VG via photo-crosslinking, and thus the oriented photo-crosslinking enzymes were obtained. The enzymatic activity, thermostability, and reusability of the affinity directional photo-crosslinked immobilized sucrose isomerase (hv-EP-VG-SI) were systematically studied. The oriented immobilization enzymes were significantly improved in recycling and heat resistance. Moreover, hv-EP-VG-SI retained more than 90% of the original activity and 50% of the activity after 11 cycles.
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