DeLorenzo Drew M, Henson William R, Moon Tae Seok
Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis , St. Louis, Missouri 63130, United States.
ACS Synth Biol. 2017 Oct 20;6(10):1973-1978. doi: 10.1021/acssynbio.7b00192. Epub 2017 Aug 10.
Rhodococcus opacus PD630 is a nonmodel, Gram-positive bacterium that possesses desirable traits for biomass conversion, including consumption capabilities for lignocellulose-based sugars and toxic lignin-derived aromatic compounds, significant triacylglycerol accumulation, relatively rapid growth rate, and genetic tractability. However, few genetic elements have been directly characterized in R. opacus, limiting its application for lignocellulose bioconversion. Here, we report the characterization and development of genetic tools for tunable gene expression in R. opacus, including: (1) six fluorescent reporters for quantifying promoter output, (2) three chemically inducible promoters for variable gene expression, and (3) two classes of metabolite sensors derived from native R. opacus promoters that detect nitrogen levels or aromatic compounds. Using these tools, we also provide insights into native aromatic consumption pathways in R. opacus. Overall, this work expands the ability to control and characterize gene expression in R. opacus for future lignocellulose-based fuel and chemical production.
嗜油红球菌PD630是一种非模式革兰氏阳性细菌，具有生物质转化所需的特性，包括对基于木质纤维素的糖类和有毒木质素衍生的芳香族化合物的消耗能力、显著的三酰甘油积累、相对较快的生长速度以及遗传易处理性。然而，在嗜油红球菌中直接表征的遗传元件很少，这限制了其在木质纤维素生物转化中的应用。在此，我们报告了嗜油红球菌中用于可调基因表达的遗传工具的表征和开发，包括：（1）六个用于量化启动子输出的荧光报告基因，（2）三个用于可变基因表达的化学诱导型启动子，以及（3）两类源自嗜油红球菌天然启动子的代谢物传感器，可检测氮水平或芳香族化合物。使用这些工具，我们还深入了解了嗜油红球菌中的天然芳香族消耗途径。总体而言，这项工作扩展了控制和表征嗜油红球菌中基因表达的能力，以用于未来基于木质纤维素的燃料和化学品生产。
