Department of Bioengineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, SW72AZ, UK.
Microb Cell Fact. 2022 Jun 14;21(1):115. doi: 10.1186/s12934-022-01842-0.
Low-cost sustainable feedstocks are essential for commercially viable biotechnologies. These feedstocks, often derived from plant or food waste, contain a multitude of different complex biomolecules which require multiple enzymes to hydrolyse and metabolise. Current standard biotechnology uses monocultures in which a single host expresses all the proteins required for the consolidated bioprocess. However, these hosts have limited capacity for expressing proteins before growth is impacted. This limitation may be overcome by utilising division of labour (DOL) in a consortium, where each member expresses a single protein of a longer degradation pathway.
Here, we model a two-strain consortium, with one strain expressing an endohydrolase and a second strain expressing an exohydrolase, for cooperative degradation of a complex substrate. Our results suggest that there is a balance between increasing expression to enhance degradation versus the burden that higher expression causes. Once a threshold of burden is reached, the consortium will consistently perform better than an equivalent single-cell monoculture.
We demonstrate that resource-aware whole-cell models can be used to predict the benefits and limitations of using consortia systems to overcome burden. Our model predicts the region of expression where DOL would be beneficial for growth on starch, which will assist in making informed design choices for this, and other, complex-substrate degradation pathways.
低成本可持续的饲料对于商业可行的生物技术至关重要。这些饲料通常来自植物或食物废物,含有多种不同的复杂生物分子,需要多种酶来水解和代谢。目前的标准生物技术使用单一培养物,其中单一宿主表达整合生物过程所需的所有蛋白质。然而,这些宿主在生长受到影响之前表达蛋白质的能力有限。这一限制可以通过在联合体中利用分工(DOL)来克服,其中每个成员表达一条较长降解途径中的单个蛋白质。
在这里,我们为一个具有两株菌的联合体建模,一株菌表达内切水解酶,另一株菌表达外切水解酶,用于合作降解复杂底物。我们的结果表明,在提高降解效率和高表达带来的负担之间存在一种平衡。一旦达到负担的阈值,联合体的性能将始终优于等效的单细胞单一培养物。
我们证明了资源感知的全细胞模型可用于预测使用联合体系统克服负担的好处和限制。我们的模型预测了 DOL 在淀粉上表达的区域,这将有助于对这种和其他复杂底物降解途径做出明智的设计选择。