Multi-Omics Analysis of Methanotroph

Methanotrophic bacterium can utilize methane, one of the next-generation chemical feedstocks, as a sole carbon source and energy to produce a variety of biofuels and chemical precursors. Recently, a novel methanotroph, Methylomonas sp. DH-1, was discovered and thought to be a promising strain as it has many strong points: relatively rapid growth than other methanotrophs, well-developed isoprenoid pathway, high ethanol tolerance, and so on. However, genetic information of this strain is not clear, which makes it difficult to take advantage of it in industries. To solve this problem, multi-omics analysis of this strain is required. In this study, we made the protocols for making various omics libraries specific to methanotrophs. Many methanotrophs have intracytoplasmic membrane where methane monooxygenase is attached. They also produce a variety of second metabolites and effuse them, which may take on a role of cushion when cells are lysed. As a result, cell lysis and total transcript extraction step is the major bottleneck of making libraries of methanotrophs. Therefore, we firstly optimized cell lysis method, and then developed protocols for RNA-seq, TSS-seq, and ribosome profiling specifically tailored to methanotrophs. We also made libraries of Methylomonas sp. DH-1 and Methylosinus trichosporium OB3b. By analyzing multi-omics data, we expect to understand the genetic information and metabolic pathways of methanotrophs well and make use of them to produce valuable products.