Stability of a Mutualistic Escherichia coli Co-Culture During Violacein Production Depends on the Kind of Carbon Source

IF 3.9 4区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Simon Schick, Tobias Müller, Ralf Takors, Georg A. Sprenger
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Abstract

The L-tryptophan–derived purple pigment violacein (VIO) is produced in recombinant bacteria and studied for its versatile applications. Microbial synthetic co-cultures are gaining more importance as efficient factories for synthesizing high-value compounds. In this work, a mutualistic and cross-feeding Escherichia coli co-culture is metabolically engineered to produce VIO. The strains are genetically modified by auxotrophies in the tryptophan (TRP) pathway to enable a metabolic division of labor. Therein, one strain produces anthranilate (ANT) and the other transforms it into TRP and further to VIO. Population dynamics and stability depend on the choice of carbon source, impacting the presence and thus exchange of metabolites as well as overall VIO productivity. Four carbon sources (D-glucose, glycerol, D-galactose, and D-xylose) were compared. D-Xylose led to co-cultures which showed stable growth and VIO production, ANT-TRP exchange, and enhanced VIO production. Best titers were ∼126 mg L–1 in shake flasks. The study demonstrates the importance and advantages of a mutualistic approach in VIO synthesis and highlights the carbon source's role in co-culture stability and productivity. Transferring this knowledge into an up-scaled bioreactor system has great potential in improving the overall VIO production.

Abstract Image

互助型大肠杆菌共培养菌群在生产维拉丝素过程中的稳定性取决于碳源种类
由 L-色氨酸衍生的紫色色素紫草素(VIO)是在重组细菌中生产出来的,其用途广泛。微生物合成共培养作为合成高价值化合物的高效工厂,正变得越来越重要。在这项工作中,通过代谢工程改造了一种互生和交叉进食的大肠杆菌共培养物,以生产 VIO。这些菌株通过色氨酸(TRP)途径中的辅助营养因子进行了基因改造,从而实现了代谢分工。其中,一个菌株生产蒽酸(ANT),另一个菌株将其转化为 TRP,再进一步转化为 VIO。种群的动态和稳定性取决于碳源的选择,这影响到代谢物的存在和交换,以及整个 VIO 的生产率。我们对四种碳源(D-葡萄糖、甘油、D-半乳糖和 D-木糖)进行了比较。D-木糖导致的共培养显示出稳定的生长和 VIO 产量、ANT-TRP 交换以及 VIO 产量的提高。在摇瓶中,最佳滴度为∼126 mg L-1。该研究证明了在 VIO 合成过程中采用互作方法的重要性和优势,并强调了碳源在共培养稳定性和生产率中的作用。将这些知识应用到大规模生物反应器系统中,对提高整体 VIO 产量具有巨大潜力。
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来源期刊
Engineering in Life Sciences
Engineering in Life Sciences 工程技术-生物工程与应用微生物
CiteScore
6.40
自引率
3.70%
发文量
81
审稿时长
3 months
期刊介绍: Engineering in Life Sciences (ELS) focuses on engineering principles and innovations in life sciences and biotechnology. Life sciences and biotechnology covered in ELS encompass the use of biomolecules (e.g. proteins/enzymes), cells (microbial, plant and mammalian origins) and biomaterials for biosynthesis, biotransformation, cell-based treatment and bio-based solutions in industrial and pharmaceutical biotechnologies as well as in biomedicine. ELS especially aims to promote interdisciplinary collaborations among biologists, biotechnologists and engineers for quantitative understanding and holistic engineering (design-built-test) of biological parts and processes in the different application areas.
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