Caseous lymphadenitis (CLA) is a chronic disease that affects small ruminants and causes economic losses in the associated breeding system. The causative agent of CLA is Corynebacterium pseudotuberculosis, a Gram-positive bacterium that exhibits tropism for external and internal lymph nodes and induces abscess formation in the host. Bacterial communities often produce a biofilm matrix that serves various functions, including protection against hostile environmental conditions, antibiotics, and the host immune response. Although biofilm formation has been reported for C. pseudotuberculosis, not all strains demonstrate this property in culture. In this work, we report the first comparative proteomic analysis of one biofilm-forming (CAPJ4) and one biofilm-non-forming strain (CAP3W) of C. pseudotuberculosis isolated from goats. Bacterial whole cell protein extracts were obtained for mass spectrometry analyses. Using LC-MS/MS, our studies reveal three and four proteins exclusively found in the CAPJ4 and CAP3W proteome, respectively. In addition, label-free quantitative analysis identified 40 proteins showing at-least 2-fold higher values in CAPJ4 compared CAP3W proteome Notably, CAPJ4 differentially synthesized the penicillin-binding protein, which participates in the formation of peptidoglycans. CAPJ4 also exhibited upregulation of N-acetylmuramoyl-L-alanine amidase and galactose-1-phosphate uridylyltransferase, which are involved in biofilm formation and exopolysaccharide biosynthesis. Here, we demonstrate that biofilm formation in C. pseudotuberculosis is likely associated with specific proteins, some of which were previously shown to be associated with virulence and biofilm formation in other organisms. Our findings may drive studies related to the bacterial mechanisms involved in the biofilm formation, in addition to providing targets for the treatment of CLA.

干酪性淋巴结炎（CLA）是一种慢性疾病，影响小反刍动物，并在相关的育种系统中造成经济损失。CLA的病因是假结核棒杆菌，是一种革兰氏阳性细菌，对外部和内部淋巴结表现出嗜性，并诱导宿主形成脓肿。细菌群落通常产生具有各种功能的生物膜基质，包括针对不利环境条件的保护，抗生素和宿主免疫反应。虽然据报道生物膜形成假结核，并非所有菌株都能在培养中证明这一特性。在这项工作中，我们报告了一个生物膜形成（CAPJ4）和一个生物膜不形成菌株（CAP3W）的第一次比较蛋白质组学分析。假结核与山羊隔离。获得细菌全细胞蛋白提取物用于质谱分析。使用LC-MS / MS，我们的研究揭示了分别在CAPJ4和CAP3W蛋白质组中独家发现的三种和四种蛋白质。此外，无标记的定量分析鉴定出40种蛋白质，与CAP3W蛋白质组相比，CAPJ4的蛋白质至少高2倍。值得注意的是，CAPJ4差异性地合成了青霉素结合蛋白，该蛋白参与了肽聚糖的形成。CAPJ4还表现出N-乙酰基muramoyl-L-丙氨酸酰胺酶和半乳糖-1-磷酸尿嘧啶转移酶的上调，它们参与生物膜的形成和胞外多糖的生物合成。在这里，我们证明了生物膜的形成假结核可能与特定蛋白质有关，某些蛋白质先前已显示与其他生物体中的毒力和生物膜形成有关。我们的发现可能会推动与生物膜形成过程中涉及的细菌机制有关的研究，并为CLA的治疗提供目标。