Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Zhejiang Shaoxing Research Institute of Tianjin University, Shaoxing 312300, China.
J Dairy Sci. 2022 Aug;105(8):6483-6498. doi: 10.3168/jds.2022-21842. Epub 2022 Jul 13.
In Lactococcus lactis, different regulation mechanisms can be activated to overcome the effects of adverse environmental stresses. Here, a TetR family regulator LssR was demonstrated as a positive regulator in the activation of the mechanisms involved in acid and nisin tolerance of L. lactis. The deletion of lssR led to the reduction of tolerance of L. lactis NZ9000 to nisin and acid stress, and the survival rates of NZ9000 under nisin and acid stress were roughly 20-fold, 10-fold (pH 3.0, hydrochloric acid), and 8.9-fold (pH 4.0, lactic acid) of the lssR mutant NZΔlssR, respectively. Moreover, the lssR mutant NZΔlssR also displayed a lower intracellular pH stability and a changed cell surface morphology. Subsequently, transcriptome analysis revealed that genes related to the arginine deiminase pathway, the surface polysaccharides biosynthesis, carbohydrates transport and metabolism, multidrug resistance, cell repair proteins and chaperones were predominantly down transcribed in NZΔlssR. The transcript levels of the arginine deiminase pathway and the surface polysaccharides biosynthesis-associated genes under acid and nisin stresses were compared between the wild type NZ9000 and NZΔlssR using real-time fluorescence quantitative PCR. It revealed that the arginine deiminase pathway genes (arcD1C1C2T) and the surface polysaccharides biosynthesis genes (cgT, gmhB, gmhA, hddA, tagH and tarS) were proposed to be the main regulatory mechanisms of LssR in response to the acid and nisin stresses. Overall, the important role of LssR in the acid and nisin stresses response was demonstrated and the putative regulation mechanism of LssR was revealed.
在乳球菌中,可以激活不同的调控机制来克服不利环境胁迫的影响。在这里,我们证明了一个 TetR 家族调控因子 LssR 是乳球菌酸和乳链菌肽耐受性机制激活的正调控因子。lssR 的缺失导致乳球菌 NZ9000 对乳链菌肽和酸胁迫的耐受性降低,NZ9000 在乳链菌肽和酸胁迫下的存活率分别约为 lssR 突变体 NZΔlssR 的 20 倍、10 倍(pH3.0、盐酸)和 8.9 倍(pH4.0、乳酸)。此外,lssR 突变体 NZΔlssR 还表现出较低的细胞内 pH 稳定性和改变的细胞表面形态。随后,转录组分析显示,与精氨酸脱亚氨酶途径、表面多糖生物合成、碳水化合物运输和代谢、多药耐药性、细胞修复蛋白和伴侣相关的基因在 NZΔlssR 中主要下调。使用实时荧光定量 PCR 比较了野生型 NZ9000 和 NZΔlssR 在酸和乳链菌肽胁迫下精氨酸脱亚氨酶途径和表面多糖生物合成相关基因的转录水平。结果表明,精氨酸脱亚氨酶途径基因(arcD1C1C2T)和表面多糖生物合成基因(cgT、gmhB、gmhA、hddA、tagH 和 tarS)可能是 LssR 响应酸和乳链菌肽胁迫的主要调控机制。总之,该研究证明了 LssR 在酸和乳链菌肽胁迫响应中的重要作用,并揭示了 LssR 的潜在调控机制。