Deng Hui, Li Sicong, Huang Yanling, Li Jiangling, Ni Qingsong, Zhao Yang, Chen Jin, Peng Xiurong, Li Bin, Yu Dan
Animal Breeding and Genetics key Laboratory of Sichuan Province, Sichuan Animal Science Academy, Chengdu, 610066, China.
Key Laboratory of Animal Science of State Ethnic Affairs Commission, College of Animal and Veterinary Sciences, Southwest Minzu University, Chengdu, 610041, China.
Mol Biol Rep. 2025 Mar 13;52(1):307. doi: 10.1007/s11033-025-10388-4.
Laccase CueO, a multicopper oxidase, possesses the capability to degrade phenolic compounds. In prior research, a strain of Escherichia coli named SDB2, isolated from chicken cecum, was found to degrade sinapine (a phenolic constituent of rapeseed meal) through the secretion of laccase CueO. Herein, the cloning, expression, and bioinformatics analysis of the CueO gene derived from E. coli SDB2 are reported.
Sequence analysis indicated that SDB2 CueO comprised 1551 bp, 516 amino acids, a putative molecular weight of 56.65 kDa, and an isoelectric point (pI) of 6.21. BLAST comparisons showed that the CueO protein sequence from E. coli SDB2 exhibited 65-90% identity with CueO from other bacteria. Multiple alignment analysis further confirmed the similarity and identity of SDB2 CueO with CueO from other species, and the amino acids surrounding the Cu-binding sites were highly conserved. A phylogenetic tree demonstrated a close evolutionary relationship between CueO from E. coli and CueO from Citrobacter amalonaticus. The three-dimensional (3D) structural model revealed four copper (Cu)-binding regions. Recombinant CueO was successfully obtained by expressing the CueO gene in E. coli BL21 after isopropyl β-D-1-thiogalactopyranoside (IPTG) induction. Bioinformatics analysis confirmed the similarity of recombinant CueO with native CueO.
These findings established a basis for understanding the characteristics and functions of laccase CueO from E. coli SDB2, paving the way for future research to explore the properties of recombinant CueO and its potential practical applications in optimizing feed resources, such as rapeseed meal, in the feed production industry.
漆酶CueO是一种多铜氧化酶，具有降解酚类化合物的能力。在先前的研究中，从鸡盲肠分离出的一株大肠杆菌SDB2被发现通过分泌漆酶CueO来降解芥子碱（菜籽粕中的一种酚类成分）。本文报道了源自大肠杆菌SDB2的CueO基因的克隆、表达及生物信息学分析。
序列分析表明，SDB2 CueO由1551个碱基对组成，有516个氨基酸，推测分子量为56.65 kDa，等电点（pI）为6.21。BLAST比较显示，大肠杆菌SDB2的CueO蛋白序列与其他细菌的CueO具有65 - 90%的同一性。多序列比对分析进一步证实了SDB2 CueO与其他物种的CueO的相似性和同一性，并且铜结合位点周围的氨基酸高度保守。系统发育树表明大肠杆菌的CueO与丙二酸柠檬酸杆菌的CueO之间存在密切的进化关系。三维（3D）结构模型显示有四个铜（Cu）结合区域。通过在异丙基β - D - 1 - 硫代半乳糖苷（IPTG）诱导后在大肠杆菌BL21中表达CueO基因，成功获得了重组CueO。生物信息学分析证实了重组CueO与天然CueO的相似性。
这些发现为理解大肠杆菌SDB2漆酶CueO的特性和功能奠定了基础，为未来研究探索重组CueO的性质及其在饲料生产行业优化饲料资源（如菜籽粕）方面的潜在实际应用铺平了道路。
