Department of Biological Sciences, University of North Texas, Denton, TX, USA.
BioDiscovery Institute, University of North Texas, Denton, TX, USA.
Methods Mol Biol. 2022;2489:421-434. doi: 10.1007/978-1-0716-2273-5_21.
Microbes with the capacity to use methane (CH) as a carbon source (methanotrophs) have significant potential for the bioconversion of CH-containing natural gas and anaerobic digestion-derived biogas to high value products. These organisms also play a vital role in the biogeochemical cycling of atmospheric CH by serving as the only known biological sink of this gas in terrestrial and aquatic ecosystems. Much is known regarding the enzymes and central metabolic pathways mediating CH utilization in these bacteria. However, large fundamental knowledge gaps exist regarding methanotroph physiology and responses to environmental stimuli, primarily due to a lack of efficient molecular tools to probe gene-function relationships. In this chapter, we describe several recently developed genetic tools and optimized genome editing methods that can be used for methanotroph metabolic engineering and to probe metabolic and physiological governing mechanisms in these unique bacteria.
能够将甲烷 (CH) 用作碳源的微生物（甲烷营养菌）具有将含 CH 的天然气和厌氧消化产生的沼气生物转化为高价值产品的巨大潜力。这些生物体还通过作为陆地和水生生态系统中大气 CH 的唯一已知生物汇，在大气 CH 的生物地球化学循环中发挥着至关重要的作用。关于这些细菌中 CH 利用所涉及的酶和中心代谢途径，人们已经有了很多了解。然而，由于缺乏有效的分子工具来探究基因功能关系，因此在甲烷营养菌生理学和对环境刺激的反应方面仍存在很大的知识空白。在本章中，我们描述了几种最近开发的遗传工具和优化的基因组编辑方法，这些方法可用于甲烷营养菌的代谢工程，并可用于探究这些独特细菌中的代谢和生理控制机制。
