Background Methane is the major component of natural and shale gas. Methane can be converted into methanol via a bioprocess using methanotrophs, and methanol is a valuable chemical feedstock for the production of value‐added chemicals. This work demonstrates highly effective bioconversion of methane to methanol using a newly isolated novel methanotroph, Methylomonas sp. DH‐1. Results A novel methanotroph strain was isolated from activated sludge from a brewery plant and characterized using phylogenetic analysis, electron microscopy and chemotaxonomic analysis. This aerobic, Gram‐negative, non‐motile rod‐shaped type I methanotroph was designated as Methylomonas sp. DH‐1. The growth condition of Methylomonas sp. DH‐1 and batch methane‐to‐methanol bioconversion conditions such as methane concentration, pH, biocatalyst loading, concentration of formate and MDH inhibitor were analyzed and optimized. Methanol was produced from methane with a 1.340 g L−1 titer, a 0.332 g L−1 h−1 volumetric conversion rate and a 0.0752 g g−1 cell h−1 specific methanol conversion rate. Conclusion It was demonstrated that isolation and application of a new methanotroph strain is a practical way of improving bioconversion efficiency in the conversion of methane to methanol. Moreover, one promising feature of Methylomonas sp. DH‐1 for methanol production was its extremely high tolerance to methanol up to 7%(v/v), which is advantageous for high‐titer methanol production. © 2016 Society of Chemical Industry

Recently, methane has attracted much attention as an alternative carbon feedstock since it is the major component of abundant shale and natural gas. In this work, we produced methanol from methane using whole cells of Methylosinus trichosporium OB3b as the biocatalyst. M. trichosporium OB3b was cultured on NMS medium with a supply of 7:3 air/methane ratio at 30°C. The optimal concentrations of various methanol dehydrogenase inhibitors such as potassium phosphate and EDTA were determined to be 100 and 0.5 mM, respectively, for an efficient production of methanol. Sodium formate (40 mM) as a reducing power source was added to enhance the conversion efficiency. A productivity of 49.0 mg/l·h, titer of 0.393 g methanol/l, and conversion of 73.8% (mol methanol/mol methane) were obtained under the optimized batch condition.

Methane‐assimilating bacteria, methanotrophs, can play an important role in producing various value‐added chemicals and biofuels from methane, which is considered a next‐generation carbon feedstock. The capability to engineer the metabolic pathway of methanotrophs is a key success factor for enhancing methane‐to‐product conversion efficiency. Recently, OMICS studies on several model methanotrophs have been conducted and provided strategies to engineer methanotrophs. Here, we present a review on the current progresses and future prospects of metabolic engineering of methanotrophs and its application to chemical and biofuel production from methane. © 2016 Society of Chemical Industry and John Wiley & Sons, Ltd

Background Methane, a main component of natural gas and biogas, has gained much attention as an abundant and low-cost carbon source. Methanotrophs, which can use methane as a sole carbon and energy source, are promising hosts to produce value-added chemicals from methane, but their metabolic engineering is still challenging. In previous attempts to produce lactic acid (LA) from methane, LA production levels were limited in part due to LA toxicity. We solved this problem by generating an LA-tolerant strain, which also contributes to understanding novel LA tolerance mechanisms. Results In this study, we engineered a methanotroph strain Methylomonas sp. DH-1 to produce d-lactic acid (d-LA) from methane. LA toxicity is one of the limiting factors for high-level production of LA. Therefore, we first performed adaptive laboratory evolution of Methylomonas sp. DH-1, generating an LA-tolerant strain JHM80. Genome sequencing of JHM80 revealed the causal gene watR, encoding a LysR-type transcription factor, whose overexpression due to a 2-bp (TT) deletion in the promoter region is partly responsible for the LA tolerance of JHM80. Overexpression of the watR gene in wild-type strain also led to an increase in LA tolerance. When d form-specific lactate dehydrogenase gene from Leuconostoc mesenteroides subsp. mesenteroides ATCC 8293 was introduced into the genome while deleting the glgA gene encoding glycogen synthase, JHM80 produced about 7.5-fold higher level of d-LA from methane than wild type, suggesting that LA tolerance is a critical limiting factor for LA production in this host. d-LA production was further enhanced by optimization of the medium, resulting in a titer of 1.19 g/L and a yield of 0.245 g/g CH4. Conclusions JHM80, an LA-tolerant strain of Methylomonas sp. DH-1, generated by adaptive laboratory evolution was effective in LA production from methane. Characterization of the mutated genes in JHM80 revealed that overexpression of the watR gene, encoding a LysR-type transcription factor, is responsible for LA tolerance. By introducing a heterologous lactate dehydrogenase gene into the genome of JHM80 strain while deleting the glgA gene, high d-LA production titer and yield were achieved from methane.

Methylomonas sp. DH-1, newly isolated from the activated sludge of a brewery plant, has been used as a promising biocatalytic platform for the conversion of methane to value-added chemicals. Methylomonas sp. DH-1 can efficiently convert methane and propane into methanol and acetone with a specific productivity of 4.31 and 0.14 mmol/g cell/h, the highest values ever reported, respectively. Here, we present the complete genome sequence of Methylomonas sp. DH-1 which consists of a 4.86 Mb chromosome and a 278 kb plasmid. The existence of a set of genes related to one-carbon metabolism and various secondary metabolite biosynthetic pathways including carotenoid pathways were identified. Interestingly, Methylomonas sp. DH-1 possesses not only the genes of the ribulose monophosphate cycle for type I methanotrophs but also the genes of the serine cycle for type II. Methylomonas sp. DH-1 accumulated 80 mM succinate from methane under aerobic conditions, because DH-1 has 2-oxoglutarate dehydrogenase activity and the ability to operate the full TCA cycle. Availability of the complete genome sequence of Methylomonas sp. DH-1 enables further investigations on the metabolic engineering of this strain for the production of value-added chemicals from methane.