Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden.
Novo Nordisk Foundation Center for Biosustainability, Chalmers University of Technology, Gothenburg, Sweden.
PLoS One. 2020 Dec 17;15(12):e0239882. doi: 10.1371/journal.pone.0239882. eCollection 2020.
Alkane-based biofuels are desirable to produce at a commercial scale as these have properties similar to current petroleum-derived transportation fuels. Rationally engineering microorganisms to produce a desirable compound, such as alkanes, is, however, challenging. Metabolic engineers are therefore increasingly implementing evolutionary engineering approaches combined with high-throughput screening tools, including metabolite biosensors, to identify productive cells. Engineering Saccharomyces cerevisiae to produce alkanes could be facilitated by using an alkane-responsive biosensor, which can potentially be developed from the native alkane-sensing system in Yarrowia lipolytica, a well-known alkane-assimilating yeast. This putative alkane-sensing system is, at least, based on three different transcription factors (TFs) named Yas1p, Yas2p and Yas3p. Although this system is not fully elucidated in Y. lipolytica, we were interested in evaluating the possibility of translating this system into an alkane-responsive biosensor in S. cerevisiae. We evaluated the alkane-sensing system in S. cerevisiae by developing one sensor based on the native Y. lipolytica ALK1 promoter and one sensor based on the native S. cerevisiae CYC1 promoter. In both systems, we found that the TFs Yas1p, Yas2p and Yas3p do not seem to act in the same way as these have been reported to do in their native host. Additional analysis of the TFs suggests that more knowledge regarding their mechanism is needed before a potential alkane-responsive sensor based on the Y. lipolytica system can be established in S. cerevisiae.
烷烃基生物燃料是理想的商业生产规模，因为它们的性质类似于当前的石油衍生运输燃料。然而，理性地设计微生物来生产所需的化合物，如烷烃，是具有挑战性的。因此，代谢工程师越来越多地采用进化工程方法，结合高通量筛选工具，包括代谢物生物传感器，来识别有生产力的细胞。通过使用烷烃响应生物传感器，可以促进酿酒酵母生产烷烃，该生物传感器可以潜在地从熟知的烷烃同化酵母解脂耶氏酵母中的天然烷烃感应系统中开发出来。这个假定的烷烃感应系统至少基于三个不同的转录因子（TFs），分别命名为 Yas1p、Yas2p 和 Yas3p。尽管这个系统在解脂耶氏酵母中没有完全阐明，但我们有兴趣评估将这个系统翻译成酿酒酵母中的烷烃响应生物传感器的可能性。我们通过开发一个基于天然解脂耶氏酵母 ALK1 启动子的传感器和一个基于天然酿酒酵母 CYC1 启动子的传感器，在酿酒酵母中评估了烷烃感应系统。在这两个系统中，我们发现 TF Yas1p、Yas2p 和 Yas3p 似乎不像在其天然宿主中那样发挥作用。对 TF 的进一步分析表明，在建立基于解脂耶氏酵母系统的潜在烷烃响应传感器之前，需要更多关于其机制的知识。
