Universität des Saarlandes, Pharmazeutische Biotechnologie, Bld. C2.3, 66123 Saarbrücken, Germany; Helmholtz-Institute for Pharmaceutical Research Saarland, UdS Campus, Bld. E8.1, 66123 Saarbrücken, Germany.
Helmholtz-Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Metab Eng. 2018 May;47:263-270. doi: 10.1016/j.ymben.2018.03.019. Epub 2018 Mar 30.
Synthetic biology techniques hold great promise for optimising the production of natural products by microorganisms. However, evaluating the phenotype of a modified bacterium represents a major bottleneck to the engineering cycle - particularly for antibiotic-producing actinobacteria strains, which grow slowly and are challenging to genetically manipulate. Here, we report the generation and application of antibiotic-specific whole-cell biosensor derived from TetR transcriptional repressor for use in identifying and optimising antibiotic producers. The constructed biosensor was successfully used to improve production of polyketide antibiotic pamamycin. However, an initial biosensor based on native genetic elements had inadequate dynamic and operating ranges. To overcome these limitations, we fine-tuned biosensor performance through alterations of the promoter and operator of output module and the ligand affinity of transcription factor module, which enabled us to deduce recommendations for building and application of actinobacterial biosensors.
合成生物学技术为优化微生物天然产物的生产提供了巨大的潜力。然而，评估经过修饰的细菌的表型是工程周期的主要瓶颈，特别是对于抗生素产生的放线菌菌株，它们生长缓慢且难以进行基因操作。在这里，我们报告了一种基于 TetR 转录抑制因子的抗生素特异性全细胞生物传感器的产生和应用，该传感器可用于鉴定和优化抗生素产生菌。所构建的生物传感器成功地用于提高聚酮抗生素帕马霉素的产量。然而，基于天然遗传元件的初始生物传感器的动态范围和工作范围不足。为了克服这些限制，我们通过改变输出模块的启动子和操作子以及转录因子模块的配体亲和力来微调生物传感器的性能，这使我们能够为放线菌生物传感器的构建和应用提出建议。
