Ho Joe C H, Pawar Sandip V, Hallam Steven J, Yadav Vikramaditya G
Department of Microbiology and Immunology, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.
Department of Chemical & Biological Engineering, University of British Columbia , Vancouver, British Columbia V6T 1Z4, Canada.
ACS Synth Biol. 2018 Feb 16;7(2):392-398. doi: 10.1021/acssynbio.7b00412. Epub 2017 Dec 4.
The discovery and utilization of biocatalysts that selectively valorize lignocellulose is critical to the profitability of next-generation biorefineries. Here, we report the development of a refactored, whole-cell, GFP-based biosensor for high-throughput identification of biocatalysts that transform lignin into specialty chemicals from environmental DNA of uncultivable archaea and bacteria. The biosensor comprises the transcriptional regulator and promoter of the emrRAB operon of E. coli, and the configuration of the biosensor was tuned with the aid of mathematical model. The biosensor sensitively and selectively detects vanillin and syringaldehyde, and responds linearly over a wide detection range. We employed the biosensor to screen 42 520 fosmid clones comprising environmental DNA isolated from two coal beds and successfully identified 147 clones that transform hardwood kraft lignin to vanillin and syringaldehyde.
选择性地将木质纤维素转化为高附加值产品的生物催化剂的发现与利用，对于下一代生物精炼厂的盈利能力至关重要。在此，我们报告了一种经过重构的、基于绿色荧光蛋白（GFP）的全细胞生物传感器的开发，用于从不可培养古菌和细菌的环境DNA中高通量鉴定将木质素转化为特种化学品的生物催化剂。该生物传感器由大肠杆菌emrRAB操纵子的转录调节因子和启动子组成，并且借助数学模型对生物传感器的结构进行了调整。该生物传感器能够灵敏且选择性地检测香草醛和紫丁香醛，并且在较宽的检测范围内呈线性响应。我们利用该生物传感器筛选了42520个包含从两个煤层分离的环境DNA的fosmid克隆，并成功鉴定出147个将阔叶木硫酸盐木质素转化为香草醛和紫丁香醛的克隆。
