Department of Biotechnology and Bioengineering, Interdisciplinary Program for Bioenergy & Biomaterials, Chonnam National University, Gwangju 61186, Republic of Korea.
Metabolic and Biomolecular Engineering National Research Laboratory, SMESH Cross-Generation Collaborative Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Plus Program), Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.
Int J Biol Macromol. 2019 Jun 15;131:29-35. doi: 10.1016/j.ijbiomac.2019.03.044. Epub 2019 Mar 7.
During microbial production of target product, accumulation of by-products and target product itself may be toxic to host strain. Thus, development of abiotic stress tolerant strains are essential to achieve high productivity of target product with sustained metabolism. Expression of DR1558 from Deinococcus radiodurans, a response regulator in two-component signal transduction system, was reported to increase the tolerance against oxidative stress in Escherichia coli. In this study, the effect of overexpression of DR1558 was examined on poly‑3‑hydroxybutyrate (PHB) production in recombinant E. coli expressing Ralstonia eutropha PHB biosynthesis genes. It was found that dr1558 overexpressing E. coli produced 5.31 g PHB/L and 9.24 g dry cell weight/L, while control strain produced 1.52 g PHB/L and 4.47 g dry cell weight/L in 48 h shake-flask cultivation. Transcriptional analysis of E. coli suggested that DR1558 could improve the expression efficiency of the genes related to central carbon metabolism and threonine bypass pathway in PHB producing E. coli. When thrABC genes were overexpressed, PHB content was increased in recombinant E. coli, which suggests that stress-tolerant genes from extremophiles should be useful in the development of engineered strains for the production of bio-based products.
在微生物生产目标产物的过程中,副产物和目标产物本身的积累可能对宿主菌株有毒。因此,开发耐非生物胁迫的菌株对于实现高目标产物生产力和持续代谢至关重要。来自耐辐射球菌的双组分信号转导系统中的应答调节剂 DR1558 的表达被报道可以提高大肠杆菌对氧化应激的耐受性。在这项研究中,研究了过表达 DR1558 对表达罗尔斯通氏菌 PHB 生物合成基因的重组大肠杆菌中聚-3-羟基丁酸(PHB)生产的影响。结果发现,dr1558 过表达大肠杆菌在 48 小时摇瓶培养中产生了 5.31 g/L PHB 和 9.24 g/L 干重,而对照菌株仅产生了 1.52 g/L PHB 和 4.47 g/L 干重。大肠杆菌的转录分析表明,DR1558 可以提高 PHB 产生大肠杆菌中与中心碳代谢和苏氨酸旁路途径相关的基因的表达效率。当 thrABC 基因过表达时,重组大肠杆菌中 PHB 含量增加,这表明来自极端微生物的耐应激基因应该有助于开发用于生产生物基产品的工程菌株。