College of Life Sciences, Nankai University, Tianjin, 300071, China.
College of Life Sciences, Nankai University, Tianjin, 300071, China; State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300071, China.
Biochem Biophys Res Commun. 2020 Mar 19;523(4):954-960. doi: 10.1016/j.bbrc.2020.01.052. Epub 2020 Jan 19.
Pseudomonas aeruginosa can metabolize acyclic monoterpenoids (such as citronellol and geraniol) as the only carbon and energy sources. A total of seven proteins (AtuA, AtuB, AtuCF, AtuD, AtuE, AtuG, AtuH) have been identified in Pseudomonas aeruginosa as participating in the acyclic terpene utilization pathway. AtuB is a dehydrogenase enzyme responsible for citronellol and geraniol catabolism in the acyclic terpene utilization (Atu) pathway, although its structure and function have not been characterized to date. Here we report the crystal structure of AtuB from Pseudomonas aeruginosa PAO1 (PaAtuB) to 1.8 Å resolution. PaAtuB crystallizes in the space group F222 with a single monomer in the asymmetric unit. Analytical ultracentrifugation data shows that PaAtuB forms a stable tetramer in solution, which is consistent with the structure. Structural analysis confirms that AtuB belongs to the short-chain dehydrogenase/reductase (SDR) family. AtuB is predicted to bind NADP(H) from the crystal structure, which is confirmed by MicroScale Thermophoresis analysis that shows PaAtuB binds NADP(H) with a Kd value of 258 μM. This work provides a starting point to explore potential biotechnology and pharmaceutical applications of AtuB.
铜绿假单胞菌可以将环状单萜（如香茅醇和香叶醇）作为唯一的碳源和能源进行代谢。铜绿假单胞菌中有七种蛋白（AtuA、AtuB、AtuCF、AtuD、AtuE、AtuG、AtuH）被鉴定为参与环状萜烯利用途径。AtuB 是一种脱氢酶，负责环状萜烯利用（Atu）途径中的香茅醇和香叶醇代谢，尽管其结构和功能尚未得到阐明。在这里，我们报道了铜绿假单胞菌 PAO1（PaAtuB）中 AtuB 的晶体结构，分辨率为 1.8Å。PaAtuB 以 F222 的空间群结晶，在不对称单元中有一个单体。分析超速离心数据表明，PaAtuB 在溶液中形成稳定的四聚体，这与结构一致。结构分析证实 AtuB 属于短链脱氢酶/还原酶（SDR）家族。从晶体结构预测 AtuB 结合 NADP(H)，这一预测得到了微量热泳动分析的证实，表明 PaAtuB 与 NADP(H)的 Kd 值为 258μM。这项工作为探索 AtuB 的潜在生物技术和制药应用提供了一个起点。
