Research Institute for Creating the Future, Fuji Oil Holdings Inc., Tsukubamirai-shi, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Japan.
FEBS Lett. 2024 Jun;598(11):1411-1421. doi: 10.1002/1873-3468.14883. Epub 2024 Apr 24.
Lipases with high interesterification activity are important enzymes for industrial use. The lipase from Burkholderia stagnalis (BsL) exhibits higher interesterification activity than that from Burkholderia plantarii (BpL) despite their significant sequence similarity. In this study, we determined the crystal structure of BsL at 1.40 Å resolution. Utilizing structural insights, we have successfully augmented the interesterification activity of BpL by over twofold. This enhancement was achieved by substituting threonine with serine at position 289 through forming an expansive space in the substrate-binding site. Additionally, we discuss the activity mechanism based on the kinetic parameters. Our study sheds light on the structural determinants of the interesterification activity of lipase.
脂肪酶具有很高的酯交换活性，是工业用途的重要酶。尽管 Burkholderia stagnalis（BsL）脂肪酶和 Burkholderia plantarii（BpL）脂肪酶的序列有显著的相似性，但 BsL 的酯交换活性却高于 BpL。在这项研究中，我们解析了 BsL 的晶体结构，分辨率为 1.40 Å。利用结构信息，我们成功地将 BpL 的酯交换活性提高了两倍以上。这是通过将 289 位的苏氨酸突变为丝氨酸，在底物结合位点形成一个扩展的空间来实现的。此外，我们还根据动力学参数讨论了其活性机制。我们的研究揭示了脂肪酶酯交换活性的结构决定因素。
