Improvement of the Process Stability of Arylmalonate Decarboxylase by Immobilization for Biocatalytic Profen Synthesis
Improvement of the Process Stability of Arylmalonate Decarboxylase by Immobilization for Biocatalytic Profen Synthesis
复制标题
DOI：
10.3389/fmicb.2017.00448
复制
发表时间：
2017-03-16
影响因子：
5.2
通讯作者：
Kara, Selin
中科院分区：
文献类型：
--
作者：
Assmann, Miriam;Muegge, Carolin;Kara, Selin
芳基丙二酸脱羧酶 (AMDase) 能够通过二羧酸前体的简单单步脱羧选择性合成对映体纯 (S)-芳基丙酸酯。然而，酶稳定性差，在工艺条件下半衰期约为 1.2 小时，严重限制了生产率，导致需要高催化剂负载。通过固定在氨基 C2 丙烯酸酯载体上，(S) 选择性 AMDase 变体 G74C/M159L/C188G/V43I/A125P/V156L 的操作稳定性增加至约 8.6 天的半衰期，这代表了 158 倍的改进。通过简单固定细胞裂解物来实现进一步的优化，以消除成本和时间密集的酶纯化步骤。
The enzyme arylmalonate decarboxylase (AMDase) enables the selective synthesis of enantiopure (S)-arylpropinates in a simple single-step decarboxylation of dicarboxylic acid precursors. However, the poor enzyme stability with a half-life time of about 1.2 h under process conditions is a serious limitation of the productivity, which results in a need for high catalyst loads. By immobilization on an amino C2 acrylate carrier the operational stability of the (S)-selective AMDase variant G74C/M159L/C188G/V43I/A125P/V156L was increased to a half-life of about 8.6 days, which represents a 158-fold improvement. Further optimization was achieved by simple immobilization of the cell lysate to eliminate the cost- and time intensive enzyme purification step.