High Cell-Density Cultivation of <i>Methylococcus capsulatus</i> Bath for Efficient Methane-Derived Mevalonate Production

Jang, Nulee; Jeong, Jiyeong; Ko, Minji; Song, Dong-Uk; Emelianov, Georgii; Kim, Seong Keun; Rha, Eugene; Kwon, Kil Koang; Kim, Haseong; Lee, Dae‐Hee; Lee, Hyewon

doi:10.1021/acs.jafc.3c00286

Cited by 8 publications

(2 citation statements)

References 44 publications

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“…Efforts have been made to improve the cell growth and productivity of methanotrophic biosynthesis by feeding nutrients or rational genetic manipulations, whereas the productivity and carbon conversion efficiency (CCE) are still major obstacles in the biomanufacturing of CH 4 -derived cell proteins (Table S1†). 24–26 Glycogen and extracellular polysaccharides (EPS) have been reported to be the predominant carbohydrates in methanotrophic bacteria, in which the significant carbon flux is split under nutritional imbalance. 19,27 Although previous studies explored strategies of nutrient exhaustion for boosting the accumulation of storage compounds from glucose, the mechanism of regulating nutritional availability for cell proteins and carbohydrates accumulated by methanotrophs using CH 4 has not been reported to date.…”

Section: Introductionmentioning

confidence: 99%

A novel nutritional induction strategy flexibly switching the biosynthesis of food-like products from methane by a methanotrophic bacterium

Gao,

Guo,

Chen

et al. 2024

Green Chem.

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Section: Introductionmentioning

confidence: 99%

A novel nutritional induction strategy flexibly switching the biosynthesis of food-like products from methane by a methanotrophic bacterium

Gao,

Guo,

Chen

et al. 2024

Green Chem.

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“…Additionally, industries such as Norferm Danmark A/S reported use of M. capsulatus Bath for the production of single cell protein, BioProtein (Bothe et al, 2002). However, efforts to optimize the culturing conditions for this bacterium are still ongoing (Jang et al, 2023).…”

Section: Introductionmentioning

confidence: 99%

Transcriptional and metabolomic responses of Methylococcus capsulatus Bath to nitrogen source and temperature downshift

Bedekar,

Deewan,

Jagtap

et al. 2023

Front. Microbiol.

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Methanotrophs play a significant role in methane oxidation, because they are the only biological methane sink present in nature. The methane monooxygenase enzyme oxidizes methane or ammonia into methanol or hydroxylamine, respectively. While much is known about central carbon metabolism in methanotrophs, far less is known about nitrogen metabolism. In this study, we investigated how Methylococcus capsulatus Bath, a methane-oxidizing bacterium, responds to nitrogen source and temperature. Batch culture experiments were conducted using nitrate or ammonium as nitrogen sources at both 37°C and 42°C. While growth rates with nitrate and ammonium were comparable at 42°C, a significant growth advantage was observed with ammonium at 37°C. Utilization of nitrate was higher at 42°C than at 37°C, especially in the first 24 h. Use of ammonium remained constant between 42°C and 37°C; however, nitrite buildup and conversion to ammonia were found to be temperature-dependent processes. We performed RNA-seq to understand the underlying molecular mechanisms, and the results revealed complex transcriptional changes in response to varying conditions. Different gene expression patterns connected to respiration, nitrate and ammonia metabolism, methane oxidation, and amino acid biosynthesis were identified using gene ontology analysis. Notably, key pathways with variable expression profiles included oxidative phosphorylation and methane and methanol oxidation. Additionally, there were transcription levels that varied for genes related to nitrogen metabolism, particularly for ammonia oxidation, nitrate reduction, and transporters. Quantitative PCR was used to validate these transcriptional changes. Analyses of intracellular metabolites revealed changes in fatty acids, amino acids, central carbon intermediates, and nitrogen bases in response to various nitrogen sources and temperatures. Overall, our results offer improved understanding of the intricate interactions between nitrogen availability, temperature, and gene expression in M. capsulatus Bath. This study enhances our understanding of microbial adaptation strategies, offering potential applications in biotechnological and environmental contexts.

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Online monitoring of methane transfer rates unveils nitrogen fixation dynamics in Methylococcus capsulatus

Engel,

Hoffmann,

Kosfeld

et al. 2024

Biotech & Bioengineering

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This study explores methane utilization by the methanotrophic microorganism Methylococcus capsulatus (Bath) for biomass production, presenting a promising approach to mitigate methane emissions and foster the development sustainable biomaterials. Traditional screening methods for gas cultivations involve either serum flasks without online monitoring or costly, low‐throughput fermenters. To address these limitations, the Respiration Activity MOnitoring System was augmented with methane sensors for real‐time methane transfer rate (MTR) monitoring in shake flasks. Utilizing online monitoring of the MTR in shake flasks results in enhanced throughput and cost‐effectiveness for cultivating M. capsulatus. Simultaneous monitoring of transfer rates for oxygen, methane, and carbon dioxide was conducted in up to eight shake flasks, ensuring the success of the cultivation process. Alterations in methane‐to‐oxygen transfer rate ratios and carbon fixation rates reveal the impact of transfer limitations on microbial growth. Detection of gas transfer limitations, exploration of process parameter influences, and investigations of medium components were enabled by the introduced method. Optimal nitrogen concentrations could be determined to ensure optimal growth. This streamlined approach accelerates the screening process, offering efficient investigations into metabolic effects, limitations, and parameter influences in gas fermentations without the need for elaborate offline sampling, significantly reducing costs and enhanced reproducibility.

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High Cell-Density Cultivation of Methylococcus capsulatus Bath for Efficient Methane-Derived Mevalonate Production

Cited by 8 publications

References 44 publications

A novel nutritional induction strategy flexibly switching the biosynthesis of food-like products from methane by a methanotrophic bacterium

A novel nutritional induction strategy flexibly switching the biosynthesis of food-like products from methane by a methanotrophic bacterium

Transcriptional and metabolomic responses of Methylococcus capsulatus Bath to nitrogen source and temperature downshift

Online monitoring of methane transfer rates unveils nitrogen fixation dynamics in Methylococcus capsulatus

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