化学
生物催化
三氟甲基
对映体
十二烷基硫酸钠
对映体过量
胺化
生物合成
共价键
产量(工程)
酶
核化学
色谱法
立体化学
有机化学
催化作用
离子液体
烷基
对映选择合成
材料科学
冶金
作者
Yuan Lü,Jinmei Wang,Pengpeng Cheng,Chuyue Zhang,Jinghua Li,Zhimin Ou
标识
DOI:10.1016/j.procbio.2023.10.003
摘要
Purified SpyCatcher-ATA117 (5 µM) and SpyTag-ADH (5 µM) were covalently combined to form self-assembled dual-enzyme clusters (DENCs) at 4 °C for 3 h. The successful assembly of DENCs was verified using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and dynamic light scattering (DLS). DENCs improved the storage stability by 36.2% and pH stability by 11.2% compared with those of free double enzymes. 56.89 g/L (R)− 1-[3,5-bis(trifluoromethyl)phenyl]ethanamine (R-BPA) was synthesized using DENCs with high yield (99.9%) and high enantiomeric excess (99.9%) at 45 °C and pH 9 for 24 h. Moreover, DENCs retained more than 78% of the initial activity after being reused in the preparation of R-BPA for five cycles. Thus, DENCs were successfully formed, and efficient amination was achieved, providing a new biocatalyst for efficient biosynthesis of chiral amines via asymmetric transamination cascade reaction.
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