| 摘要: |
| 【目的】解析施氏假单胞菌（Pseudomonas stutzeri）A1501中LuxR-0394蛋白的结构特征及其在群体感应相关表型中的潜在作用，为开发以群体感应为调控靶点的农业微生物应用提供参考。【方法】采用生物信息学方法，分析施氏假单胞菌A1501菌株中LuxR-0394蛋白的理化性质、结构域、亲疏水性、磷酸化位点、亚细胞定位、结合口袋、系统发育关系及蛋白互作网络 ，构建突变株LuxR-0394和回补株C-0394，并分别对其进行生长曲线、生物膜形成及运动能力测定。【结果】LuxR-0394蛋白缺乏典型的AHL结合结构域，保留HTH_LuxR DNA结合结构域，为LuxR solos型调控蛋白。该蛋白由218个氨基酸组成，定位于细胞质，无信号肽和跨膜结构，存在8个磷酸化位点及2个潜在小分子结合口袋。系统发育分析结果显示，LuxR-0394蛋白与P. stutzeri RCH2 同源蛋白亲缘性较近。蛋白互作分析推测其可能与双组分系统及表面结构装配相关蛋白协同调控。功能测定结果表明，PST_0394 基因缺失虽不影响施氏假单胞菌A1501生长，但导致其生物膜形成能力下降69.14%，运动能力下降87.70%。【结论】LuxR-0394在施氏假单胞菌A1501生物膜形成和运动能力等与环境适应相关的表型中发挥重要作用，并可能通过与双组分系统及表面结构装配相关蛋白的互作来影响菌株的生态功能。 |
| 关键词: 施氏假单胞菌 生物膜 运动能力 群体感应 孤立型LuxR |
| DOI：10. 13207/j. jnwafu. 2026. 10. 017 |
| 分类号: |
| 基金项目:国家自然科学基金项目（31800070）；四川省科技厅自然科学基金项目（2022NSFSC1679）；成都医学院科技项目（CYZYB-25-15） |
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| Structural characterization and functional exploration of LuxR-0394 in Pseudomonas stutzeri |
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HAN Yunlei1, HU Qi2, YOU Jiawei3, HUANG Dan2, MENG Wenjiao2, LUO Rui2, LU Jing4, TANG Ke2
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1.School of Basic Medical Sciences，Chengdu Medical College，Chengdu，Sichuan 610500，China;2.School of Public Health，Chengdu Medical College，Chengdu，Sichuan 610500，China;3.College of Bioscience and Technology，Chengdu Medical College，Chengdu，Sichuan 610500，China;4.School of Pharmaceutical Sciences，Chengdu Medical College，Chengdu，Sichuan 610500，China
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| Abstract: |
| 【Objective】This study aimed to characterize the structural features and elucidate the potential role of the LuxR solo protein LuxR-0394 in Pseudomonas stutzeri A1501，particularly in quorum sensing（QS）-related phenotypes，thus providing a foundation for the development of QS-based regulatory strategies in agricul?tural microbial applications.【Method】Bioinformatics tools were used to analyze physicochemical properties，structural domains，hydropathicity，phosphorylation sites，subcellular localization，binding pockets，phylogenetic relationships，and protein interaction networks of the LuxR-0394 protein in P.stutzeri A1501. In addition，LuxR-0394 mutant and complemented strains（C-0394）were constructed，and their growth curves，biofilm formation，and motility were subsequently evaluated.【Result】LuxR-0394 lacks the canonical AHL-binding domain but retains the HTH_LuxR DNA-binding domain，making it a LuxR solo-type regulator protein. This 218-amino-acid protein localizes in the cytoplasm without a signal peptide or transmembrane region，and is predicted to possess eight phosphorylation sites and two potential small-molecule binding pockets. Phylogenetic analysis showed its close relatedness to a homolog in P.stutzeri RCH2. Interaction network analysis suggested its coope-ration with proteins involved in two-component systems and surface structure assembly. Deletion of PST_0394 did not affect bacterial growth but significantly reduced biofilm formation （69.14%，P<0.05） and motility（87.70%，P<0.05）.【Conclusion】LuxR-0394 contributes critically to environmental adaptation-related phe?
notypes，including biofilm formation and motility，in P.stutzeri A1501，and may modulate the strain’s ecological functions through interactions with proteins associated with two-component systems and surface structure assem-bly. |
| Key words: Pseudomonas stutzeri biofilm motility quorum sensing LuxR solo |
