College of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, Anhui Province, PR China.
College of Food and Biological Engineering, Hefei University of Technology, Hefei 230009, Anhui Province, PR China.
Bioorg Chem. 2024 Jun;147:107382. doi: 10.1016/j.bioorg.2024.107382. Epub 2024 Apr 16.
Amino acid transferase is a family of enzymes used to catalyze and separate chiral amino acids. However, due to the low efficiency, by-products and reverse reactions occur in cascade reactions. Therefore, in the research, phenylglycine aminotransferase and aspartate aminotransferase were self-assembled in vitro by leucine zipper. The self-assembled enzyme system with d-phenylglycine and α-ketoglutarate as substrates were used for the chiral transformation reaction. By studying the enzyme combination, kinetic reaction stability and catalytic efficiency, it was found that the self-assembled enzyme showed improved stability and better affinity to the substrate than the control and achieved only ee value of 17.86% for the control at the substrate ratio was 1:2. In contrast, the self-assembled enzyme basically catalyzed the complete conversion of d-Phg to l-Phg, with the ee value as 99%. These results demonstrated the feasibility of the leucine zipper and the conversion of d-phenylglycine to the l-type by fusion enzyme.
氨基酸转移酶是一类用于催化和分离手性氨基酸的酶。然而，由于效率低，级联反应中会发生副产物和逆反应。因此，在研究中，通过亮氨酸拉链将苯丙氨酸氨基转移酶和天冬氨酸氨基转移酶在体外自组装。以 d-苯甘氨酸和α-酮戊二酸为底物的自组装酶系统用于手性转化反应。通过研究酶的组合、动力学反应稳定性和催化效率，发现自组装酶的稳定性得到提高，对底物的亲和力更好，在底物比为 1:2 时，控制组的 ee 值仅为 17.86%，而自组装酶基本催化 d-Phg 完全转化为 l-Phg，ee 值为 99%。这些结果表明亮氨酸拉链和融合酶将 d-苯甘氨酸转化为 l-型的可行性。
