Faculty of Natural System, Institute of Science and Engineering, Kanazawa University, Kanazawa, Ishikawa, Japan.
Research Institute of Innovative Technology for the Earth, Kizugawa, Kyoto, Japan.
Appl Microbiol Biotechnol. 2020 Aug;104(15):6719-6729. doi: 10.1007/s00253-020-10739-0. Epub 2020 Jun 17.
Cell proliferation is achieved through numerous enzyme reactions. Temperature governs the activity of each enzyme, ultimately determining the optimal growth temperature. The synthesis of useful chemicals and fuels utilizes a fraction of available metabolic pathways, primarily central metabolic pathways including glycolysis and the tricarboxylic acid cycle. However, it remains unclear whether the optimal temperature for these pathways is correlated with that for cell proliferation. Here, we found that wild-type Corynebacterium glutamicum displayed increased glycolytic activity under non-growing anaerobic conditions at 42.5 °C, at which cells do not proliferate under aerobic conditions. At this temperature, glucose consumption was not inhibited and increased by 28% compared with that at the optimal growth temperature of 30 °C. Transcriptional analysis revealed that a gene encoding glucose transporter (iolT2) was upregulated by 12.3-fold compared with that at 30 °C, with concomitant upregulation of NCgl2954 encoding the iolT2-regulating transcription factor. Deletion of iolT2 decreased glucose consumption rate at 42.5 °C by 28%. Complementation of iolT2 restored glucose consumption rate, highlighting the involvement of iolT2 in the accelerating glucose consumption at an elevated temperature. This study shows that the optimal temperature for glucose metabolism in C. glutamicum under anaerobic conditions differs greatly from that for cell growth under aerobic conditions, being beyond the upper limit of the growth temperature. This is beneficial for fuel and chemical production not only in terms of increasing productivity but also for saving cooling costs. KEY POINTS: • C. glutamicum accelerated anaerobic glucose consumption at elevated temperature. • The optimal temperature for glucose consumption was above the upper limit for growth. • Gene expression involved in glucose transport was upregulated at elevated temperature. Graphical abstract.
细胞增殖是通过许多酶反应实现的。温度控制着每种酶的活性,最终决定了最佳生长温度。有用化学品和燃料的合成利用了一部分可用的代谢途径,主要是包括糖酵解和三羧酸循环在内的中心代谢途径。然而,目前尚不清楚这些途径的最佳温度是否与细胞增殖的最佳温度相关。在这里,我们发现野生型谷氨酸棒杆菌在 42.5°C 的非生长厌氧条件下表现出增强的糖酵解活性,而在有氧条件下,细胞在此温度下不会增殖。在这个温度下,葡萄糖消耗没有受到抑制,与最佳生长温度 30°C 相比增加了 28%。转录分析表明,与 30°C 相比,编码葡萄糖转运蛋白(iolT2)的基因上调了 12.3 倍,同时调节转录因子 NCgl2954 编码的 iolT2 也上调了。iolT2 缺失使 42.5°C 时的葡萄糖消耗速率降低了 28%。iolT2 的互补恢复了葡萄糖消耗速率,突出了 iolT2 在高温下加速葡萄糖消耗的作用。本研究表明,在厌氧条件下,谷氨酸棒杆菌葡萄糖代谢的最佳温度与有氧条件下细胞生长的最佳温度有很大差异,超出了生长温度的上限。这不仅有利于提高生产力,而且有利于节约冷却成本,有利于燃料和化学品的生产。关键点:•C. glutamicum 在高温下加速了厌氧葡萄糖的消耗。•葡萄糖消耗的最佳温度高于生长的上限。•在高温下,参与葡萄糖转运的基因表达上调。