恶臭假单胞菌KT2440在内源或外源压力下表现出强大的耐受性。冷休克蛋白在极端条件下能够维持细胞的结构和功能稳定。为深入研究在有机化合物存在下细胞的耐受性,基于恶臭假单胞菌KT2440构建了敲除和过表达冷休克蛋白CspAⅠ、CspAⅡ的菌株。在紫苏醇的胁迫下,通过分光光度计和HPLC检测了细胞生长状态、半抑制浓度以及紫苏醇的产量。实验结果显示,在紫苏醇胁迫的情况下,过表达CspAⅠ的细胞的生长状态较好,其OD600值为2.57,且回补CspAⅠ菌株的半抑制浓度是敲除菌株的1.88倍。此外,在外源添加柠檬烯催化产生紫苏醇的实验中,过表达CspAⅠ的细胞展现出最佳的生长状态,且产生的紫苏醇达到了156.49 mg/L。该结果表明冷休克蛋白的过表达有助于提高细胞对有机化合物的生存能力,并且稳定了细胞代谢水平。
Pseudomonas putida KT2440 demonstrates robust tolerance under both endogenous and exogenous pressures.Cold shock proteins maintain cellular structure and function stability under extreme conditions.To delve deeper into the cells tolerance under the presence of organic compound, this study engineered strains of Pseudomonas putida KT2440 with knockout and overexpression of cold shock proteins CspAⅠand CspAⅡ.Under perilla alcohol stress, this study assessed cell growth status, half-maximal inhibitory concentration, and thymol production using a spectrophotometer and HPLC.Experimental results revealed that, under perilla alcohol stress, cells overexpressing CspAⅠshowed improved growth with an OD600 of 2.57.The half-maximal inhibitory concentration for the complemented CspAⅠstrain was 1.88 times that of the knockout strain.Furthermore, in experiments where exogenous addition of limonene catalyzed the production of perilla alcohol, cells overexpressing CspAⅠdisplayed optimal growth status, producing perilla alcohol at a level of 156.49 mg/L.This suggested that the overexpression of cold shock proteins contributes to enhancing the cells survival capability against organic compounds and stabilizing cellular metabolic levels.
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