School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, China.
School of Biological Engineering, Dalian Polytechnic University, Dalian, 116034, China.
Protein Expr Purif. 2024 Feb;214:106376. doi: 10.1016/j.pep.2023.106376. Epub 2023 Oct 14.
The novel cold-adapted lipase (Lip ZC12) derived from Psychrobacter sp. ZY124 exhibited higher catalytic activity at 20-40 °C, the whole gene was then sequenced, analyzed, and overexpressed. However, its intrinsic structural characteristics lead to a decreased affinity toward the substrate, thus limiting the improvement of catalytic efficiency. Modeling the homologous structure and simulating the binding process of Lip ZC12 with the substrate. It was found that truncated lid (lip-Δlid) could not only increase the k, but also significantly enhance the substrate affinity, the substrate affinity and catalytic efficiency of Lip ZC12 modified by lid truncation were significantly improved. The results revealed that the k/K value of lip-Δlid was 1.6 times higher than that of free lipase. This improved catalytic performance of cold-adapted lipase, and these findings laid an important foundation for further application.
新型低温适应脂肪酶（Lip ZC12）来源于Psychrobacter sp. ZY124，在 20-40°C 时表现出更高的催化活性，随后对整个基因进行了测序、分析和过表达。然而，其内在的结构特征导致其对底物的亲和力降低，从而限制了催化效率的提高。通过模拟同源结构并模拟 Lip ZC12 与底物的结合过程，发现截短的盖子（lip-Δlid）不仅可以提高 kcat，还可以显著提高底物亲和力，经盖子截短修饰后的 Lip ZC12 的底物亲和力和催化效率得到显著提高。结果表明，lip-Δlid 的 kcat/Km 值比游离脂肪酶高 1.6 倍。这种改进的低温适应脂肪酶的催化性能为进一步的应用奠定了重要基础。
