School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing, China.
Department of Bioengineering, Imperial College London, London, United Kingdom.
Appl Environ Microbiol. 2019 Mar 22;85(7). doi: 10.1128/AEM.02560-18. Print 2019 Apr 1.
DSM 743B offers potential as a chassis strain for biomass refining by consolidated bioprocessing (CBP). However, its -butanol production from lignocellulosic biomass has yet to be demonstrated. This study demonstrates the construction of a coenzyme A (CoA)-dependent acetone-butanol-ethanol (ABE) pathway in by introducing and genes from ATCC 824, which enabled it to produce -butanol using the abundant and low-cost agricultural waste of alkali-extracted, deshelled corn cobs (AECC) as the sole carbon source. Then, a novel adaptive laboratory evolution (ALE) approach was adapted to strengthen the -butanol tolerance of to fully utilize its -butanol output potential. To further improve -butanol production, both metabolic engineering and evolutionary engineering were combined, using the evolved strain as a host for metabolic engineering. The -butanol production from AECC of the engineered was increased 138-fold, from less than 0.025 g/liter to 3.47 g/liter. This method represents a milestone toward -butanol production by CBP, using a single recombinant clostridium strain. The engineered strain offers a promising CBP-enabling microbial chassis for -butanol fermentation from lignocellulose. Due to a lack of genetic tools, DSM 743B has not been comprehensively explored as a putative strain platform for -butanol production by consolidated bioprocessing (CBP). Based on the previous study of genetic tools, strain engineering of for the development of a CBP-enabling microbial chassis was demonstrated in this study. Metabolic engineering and evolutionary engineering were integrated to improve the -butanol production of from the low-cost renewable agricultural waste of alkali-extracted, deshelled corn cobs (AECC). The -butanol production from AECC was increased 138-fold, from less than 0.025 g/liter to 3.47 g/liter, which represents the highest titer of -butanol produced using a single recombinant clostridium strain by CBP reported to date. This engineered strain serves as a promising chassis for -butanol production from lignocellulose by CBP.
DSM 743B 作为一种底盘菌株，通过整合生物加工（CBP）具有生物量精炼的潜力。然而，它从木质纤维素生物质生产正丁醇的能力尚未得到证明。本研究通过引入来自 ATCC 824 的辅酶 A（CoA）依赖性丙酮丁醇乙醇（ABE）途径的 和 基因，在 中构建了该途径，从而使其能够使用丰富且廉价的农业废弃物碱提取脱壳玉米芯（AECC）作为唯一的碳源生产正丁醇。然后，采用一种新的适应性实验室进化（ALE）方法来增强 对正丁醇的耐受性，以充分利用其正丁醇生产潜力。为了进一步提高正丁醇的产量，采用代谢工程和进化工程相结合的方法，利用进化后的菌株作为宿主进行代谢工程。通过代谢工程，工程菌从 AECC 中生产的正丁醇产量增加了 138 倍，从不到 0.025g/L 增加到 3.47g/L。这种方法代表了使用单一重组梭菌菌株通过 CBP 生产正丁醇的一个里程碑。该工程菌株为木质纤维素通过 CBP 发酵生产正丁醇提供了一种很有前途的 CBP 使能微生物底盘。由于缺乏遗传工具，DSM 743B 尚未作为通过整合生物加工（CBP）生产正丁醇的潜在菌株平台进行全面探索。基于先前对遗传工具的研究，本研究中展示了对 进行菌株工程改造，以开发 CBP 使能微生物底盘。本研究整合了代谢工程和进化工程，以提高 从低成本可再生农业废弃物碱提取脱壳玉米芯（AECC）中生产正丁醇的能力。AECC 生产正丁醇的产量增加了 138 倍，从不到 0.025g/L 增加到 3.47g/L，这是迄今为止报道的通过 CBP 用单个重组梭菌菌株生产的最高正丁醇浓度。该工程菌株为通过 CBP 从木质纤维素生产正丁醇提供了一个很有前途的底盘。
