基于异腈的多组分反应在多壁碳纳米管上固定化脂肪酶：用于外消旋布洛芬的动力学拆分

Immobilization of lipase through isocyanide-based multi component reaction on multi-walled carbon nanotube: application for kinetic resolution of rac-ibuprofen.

作者信息

Moguei Mohamad Reza Safarpoor, Habibi Zohreh, Shahedi Mansour, Yousefi Maryam, Alimoradi Abouzar, Mobini Sepideh, Mohammadi Mehdi

机构信息

Department of Organic Chemistry and Oil, Faculty of Chemistry, Shahid Beheshti University, Tehran, Iran.

Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.

出版信息

Biotechnol Rep (Amst). 2022 Aug 12;35:e00759. doi: 10.1016/j.btre.2022.e00759. eCollection 2022 Sep.

lipase (TLL) was immobilized on epoxy functionalized hydroxyl multi-walled carbon nanotube (MWCNT-OH) via an isocyanide-based multicomponent reaction. The immobilization process was carried out in deionized water (pH 7.0) at room temperature resulting in loading of 600 mg enzyme/g of support with specific activity 16.9 U/mg. An immobilization yield of 100% was obtained with the expressed activity 60%. The immobilized preparation exhibited an increased thermal stability with 49% residual activity at 75 °C compared with 19% for the free enzyme at the same temperature. Solvent stability in a high ratio of DMSO was improved from 52% in free TLL to 75% in immobilized TLL. The immobilized preparation was used for kinetic resolution of rac-ibuprofen through esterification of ibuprofen in isooctane as solvent. The best result was obtained with ethanol at 45 °C and molar ratio of 2.5:1 ethanol:ibuprofen in 1 ml isooctane with 99% ee and E-value 300.

脂肪酶（TLL）通过异腈基多组分反应固定在环氧官能化的羟基多壁碳纳米管（MWCNT-OH）上。固定化过程在室温下于去离子水（pH 7.0）中进行，得到的负载量为每克载体600毫克酶，比活性为16.9 U/毫克。固定化产率为100%，表达活性为60%。固定化制剂表现出更高的热稳定性，在75°C时残留活性为49%，而游离酶在相同温度下为19%。在高比例二甲基亚砜中的溶剂稳定性从游离TLL的52%提高到固定化TLL的75%。固定化制剂用于通过在异辛烷作为溶剂中对布洛芬进行酯化来动力学拆分外消旋布洛芬。在45°C、乙醇与布洛芬摩尔比为2.5:1、体积为1毫升异辛烷的条件下使用乙醇，得到了最佳结果，对映体过量值（ee）为99%，E值为300。