恶臭假单胞菌KT2440是一种代谢通用的,HV1认证,基因可及,因此,用于生物技术应用的有趣的微生物底盘。然而,其专性需氧性质阻碍了氧敏感产品的生产,并提高了大规模发酵的成本。不能进行厌氧发酵已归因于ATP产生不足和在这些条件下不能产生嘧啶。解决这些瓶颈可以在微氧条件下生长,但在缺氧条件下不会导致生长或存活。
这里,使用数据驱动的方法来开发能够厌氧呼吸的P.putidaKT2440衍生菌株的合理设计。来到设计,将来自1628株假单胞菌菌株基因组比较的数据与基因组规模代谢模型模拟和一个包含47个样本的转录组数据集结合起来,这些样本代表兼性厌氧菌铜绿假单胞菌的14种环境条件.
结果表明,在P.putidaKT2440中实施厌氧呼吸将需要至少49个已知功能的额外基因,至少8个基因编码未知功能的蛋白质,和3种外部添加的维生素。
Pseudomonas putida KT2440 is a metabolically versatile, HV1-certified, genetically accessible, and thus interesting microbial chassis for biotechnological applications. However, its obligate aerobic nature hampers production of oxygen sensitive products and drives up costs in large scale fermentation. The inability to perform anaerobic fermentation has been attributed to insufficient ATP production and an inability to produce pyrimidines under these conditions. Addressing these bottlenecks enabled growth under micro-oxic conditions but does not lead to growth or survival under anoxic conditions.
Here, a data-driven approach was used to develop a rational design for a P. putida KT2440 derivative strain capable of anaerobic respiration. To come to the design, data derived from a genome comparison of 1628 Pseudomonas strains was combined with genome-scale metabolic modelling simulations and a transcriptome dataset of 47 samples representing 14 environmental conditions from the facultative anaerobe Pseudomonas aeruginosa.
The results indicate that the implementation of anaerobic respiration in P. putida KT2440 would require at least 49 additional genes of known function, at least 8 genes encoding proteins of unknown function, and 3 externally added vitamins.