CAS Key Laboratory of Microbial Physiological and Metabolic Engineering, State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
Biochem Biophys Res Commun. 2021 Apr 30;551:71-77. doi: 10.1016/j.bbrc.2021.03.002. Epub 2021 Mar 12.
Glyoxylate is an important chemical and is also an intermediate involved in metabolic pathways of living microorganisms. However, it cannot be rapidly utilized by many microbes. We observed a very long lag phase (up to 120 h) when E. coli is growing in a mineral medium supplemented with 50 mM glyoxylate. To better understand this strange growth pattern on glyoxylate and accelerate glyoxylate utilization, a random genomic library of E. coli was transformed into E. coli BW25113, and mutants that showed significantly shortened lag phase on glyoxylate were obtained. Interestingly, mutations in BtsT/BtsS, a two component system that is involved in pyruvate transport, were found to be a common feature in all mutants retrieved. We further demonstrated, through reverse engineering, that the mutations in BtsT/BtsS can indeed increase glyoxylate uptake. Growth experiments with different concentration of glyoxylate also showed the higher the concentration of glyoxylate, the shorter the lag phase. These new findings thus increased our understanding on microbial utilization of glyoxylate.
乙醛酸是一种重要的化学物质,也是生物体内代谢途径所涉及的中间产物。然而,许多微生物并不能快速利用它。当大肠杆菌在添加 50mM 乙醛酸的矿物培养基中生长时,我们观察到一个非常长的延迟期(长达 120 小时)。为了更好地理解乙醛酸上这种奇怪的生长模式并加速乙醛酸的利用,我们将大肠杆菌的随机基因组文库转化为大肠杆菌 BW25113,并获得了在乙醛酸上表现出明显缩短延迟期的突变体。有趣的是,在涉及丙酮酸运输的双组分系统 BtsT/BtsS 中的突变被发现是所有回收的突变体的共同特征。我们通过反向工程进一步证明,BtsT/BtsS 中的突变确实可以增加乙醛酸的摄取。不同浓度乙醛酸的生长实验也表明,乙醛酸的浓度越高,延迟期越短。这些新发现增加了我们对微生物利用乙醛酸的理解。