当前位置：
X-MOL 学术
›
ChemCatChem
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Smart Nanoparticles for Selective Immobilization of Acid Phosphatases
ChemCatChem ( IF 3.8 ) Pub Date : 2018-07-17 , DOI: 10.1002/cctc.201800405 Flóra Nagy 1 , Gábor Tasnádi 2, 3 , Diána Balogh-Weiser 1 , Evelin Bell 1 , Mélanie Hall 3 , Kurt Faber 3 , László Poppe 1, 4
ChemCatChem ( IF 3.8 ) Pub Date : 2018-07-17 , DOI: 10.1002/cctc.201800405 Flóra Nagy 1 , Gábor Tasnádi 2, 3 , Diána Balogh-Weiser 1 , Evelin Bell 1 , Mélanie Hall 3 , Kurt Faber 3 , László Poppe 1, 4
Affiliation
|
An easy to use method combining the selectivity of metal chelate affinity binding with strong covalent linking was developed for immobilization of non‐specific acid phosphatases bearing a His‐tag from crude cell lysate. Silica nanoparticles were grafted with aminopropyl functions which were partially transformed further with EDTA dianhydride to chelators. The heterofunctionalized nanoparticles charged with Ni2+ as the most appropriate metal ion were applied as support. First, the His‐tagged phosphatases were selectively bound to the metal‐chelate functions of the support. Then, the enzyme‐charged silica nanoparticles were further stabilized by forming a covalent linkage between nucleophilic moieties at the enzyme surface and free amino groups of the support using neopentylglycol diglycidylether as the most effective bifunctional linking agent. The phosphatase biocatalysts obtained by this method exhibited better phosphate transfer activity with a range of alcohols and PPi as phosphate donor in aqueous medium applying batch and continuous‐flow modes than the ones immobilized on conventional supports. Furthermore, this novel strategy opens up novel possibility for efficient immobilization of other His‐tagged recombinant enzymes.
中文翻译：
用于选择性固定酸性磷酸酶的智能纳米颗粒
开发了一种易于使用的方法，将金属螯合物亲和结合的选择性与强共价连接相结合，用于固定来自粗细胞裂解液的带有组氨酸标签的非特异性酸性磷酸酶。二氧化硅纳米粒子接枝有氨丙基官能团，其部分用 EDTA 二酐进一步转化为螯合剂。负载Ni 2+作为最合适的金属离子的异功能化纳米粒子被用作载体。首先，组氨酸标记的磷酸酶选择性地与载体的金属螯合功能结合。然后，使用新戊二醇二缩水甘油醚作为最有效的双功能连接剂，通过在酶表面的亲核部分和载体的游离氨基之间形成共价键，进一步稳定带酶的二氧化硅纳米颗粒。通过该方法获得的磷酸酶生物催化剂在水性介质中以一系列醇和 PP i作为磷酸盐供体，采用间歇和连续流动模式，表现出比固定在传统载体上的磷酸酶生物催化剂更好的磷酸盐转移活性。此外，这种新策略为有效固定其他组氨酸标签重组酶开辟了新的可能性。
更新日期：2018-07-17
中文翻译：
用于选择性固定酸性磷酸酶的智能纳米颗粒
开发了一种易于使用的方法，将金属螯合物亲和结合的选择性与强共价连接相结合，用于固定来自粗细胞裂解液的带有组氨酸标签的非特异性酸性磷酸酶。二氧化硅纳米粒子接枝有氨丙基官能团，其部分用 EDTA 二酐进一步转化为螯合剂。负载Ni 2+作为最合适的金属离子的异功能化纳米粒子被用作载体。首先，组氨酸标记的磷酸酶选择性地与载体的金属螯合功能结合。然后，使用新戊二醇二缩水甘油醚作为最有效的双功能连接剂，通过在酶表面的亲核部分和载体的游离氨基之间形成共价键，进一步稳定带酶的二氧化硅纳米颗粒。通过该方法获得的磷酸酶生物催化剂在水性介质中以一系列醇和 PP i作为磷酸盐供体，采用间歇和连续流动模式，表现出比固定在传统载体上的磷酸酶生物催化剂更好的磷酸盐转移活性。此外，这种新策略为有效固定其他组氨酸标签重组酶开辟了新的可能性。
京公网安备 11010802027423号