Potassium ion leakage impairs thermotolerance in Corynebacterium glutamicum.

Corynebacterium glutamicum, a gram-positive bacterium, can produce amino acids such as glutamic acid and lysine. The heat generated during cell growth and/or glutamate fermentation disturbs both the cell growth and fermentation. To overcome such a negative effect of the fermentation heat, we have tried to establish a high temperature fermentation. One of the approach is to create a thermotolerant strains, while the other is to create an optimum culture conditions able for the strain to grow at higher temperatures. In this study, we focused on the latter approach, where we examined the effect of potassium ion on cell growth at high growth temperatures of C. glutamicum. The supplementation of high concentrations of potassium chloride (300 mM) (or sorbitol, an osmolyte) mitigated the repressed cell growth induced by high temperature at 39 °C or 40 °C. The intracellular potassium concentration declines from 300 mM to ∼150 mM by increasing the growth temperature but not by supplementing potassium chloride or sorbitol. Furthermore, in vitro experiments revealed that the potassium ion leakage occurs at high temperatures, which was mitigated in the presence of high concentrations of extracellular potassium chloride. This suggested that the presence of high osmolyte in the culture medium could inhibit the potassium ion leakage induced by high temperature and subsequently support cell growth at high temperatures.

The effect of reactive oxygen species (ROS) and ROS-scavenging enzymes, superoxide dismutase and catalase, on the thermotolerant ability of Corynebacterium glutamicum.

The function of two reactive oxygen species (ROS) scavenging enzymes, superoxide dismutase (SOD) and catalase, on the thermotolerant ability of Corynebacterium glutamicum was investigated. In this study, the elevation of the growth temperature was shown to lead an increased intracellular ROS for two strains of Corynebacterium glutamicum, the wild-type (KY9002) and the temperature-sensitive mutant (KY9714). In order to examine the effects of ROS-scavenging enzymes on cell growth, either the SOD or the catalase gene was disrupted or overexpressed in KY9002 and KY9714. In the case of the KY9714 strain, it was shown that the disruption of SOD and catalase disturbs cell growth, while the over-productions of both the enzymes enhances cell growth with a growth temperature of 30 °C and 33 °C. Whereas, in the relatively thermotolerant KY9002 strain, the disruption of both enzymes exhibited growth defects more intensively at higher growth temperatures (37 °C or 39 °C), while the overexpression of at least SOD enhanced the cell growth at higher temperatures. Based on the correlation between the cell growth and ROS level, it was suggested that impairment of cell growth in SOD or catalase-disrupted strains could be a result of an increased ROS level. In contrast, the improvement in cell growth for strains with overexpressed SOD or catalase resulted from a decrease in the ROS level, especially at higher growth temperatures. Thus, SOD and catalase might play a crucial role in the thermotolerant ability of C. glutamicum by reducing ROS-induced temperature stress from higher growth temperatures.