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Imbalanced metabolism induced NH4+ accumulation and its effect on the central metabolism of Methylomonas sp. ZR1

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Abstract

Nitrogen and carbon are the two most essential nutrient elements, and their metabolism is tightly coupled in single carbon metabolic microorganisms. However, the nitrogen metabolism and the nitrogen/carbon (N/C) metabolic balance in single-carbon metabolism is poorly studied. In this study, the nitrogen metabolism pattern of the fast growing methanotrophs Methylomonas sp. ZR1 grown in methane and methanol was studied. Effect study of different nitrogen sources on the cell growth of ZR1 indicates that nitrate salts are the best nitrogen source supporting the growth of ZR1 using methane and methanol as carbon source. However, its metabolic intermediate ammonium was found to accumulate with high N/C ratio in the medium and consequently inhibit the growth of ZR1. Studies of carbon and nitrogen metabolic kinetic under different N/C ratio conditions indicate that the accumulation of NH4+ is caused by the imbalanced nitrogen and carbon metabolism in ZR1. Feeding carbon skeleton α-ketoglutaric acid could effectively relieve the inhibition effect of NH4+ on the growth of ZR1, which further confirms this assumption. qPCR analysis of the expression level of the central metabolic key enzyme gene indicates that the nitrogen metabolic intermediate ammonium has strong regulation effect on the central nitrogen and carbon metabolism in ZR1. qPCR-combined genomic analysis confirms that a third ammonium assimilation pathway glycine synthesis system is operated in ZR1 to balance the nitrogen and carbon metabolism. Based on the qPCR result, it was also found that ZR1 employs two strategies to relieve ammonium stress in the presence of ammonium: assimilating excess ammonium or cutting off the nitrogen reduction reactions according to the available C1 substrate. Validating the connections between single-carbon and nitrogen metabolism and studying the accumulation and assimilation mechanism of ammonium will contribute to understand how nitrogen regulates cellular growth in single-carbon metabolic microorganisms.

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Data is available for request.

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Funding

This work was supported by the National Key R&D Program of China (2021YFC2103500), STS Project of the Chinese Academy of Sciences (No. KFJ-STS-ZDTP-065), and the Key Project of Tianjin (No. 14ZCZDSY00157).

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Authors and Affiliations

  1. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Science, 32 West 7Th Avenue, Tianjin Airport Economic Area, Tianjin, 300308, China

    Wei Guo, Ronglin He, Yujie Zhao & Demao Li

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  1. Wei Guo
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  2. Ronglin He
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  3. Yujie Zhao
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  4. Demao Li
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Contributions

WG contributed to the study conception and design. Material preparation, data collection, and analysis were performed by WG and YZ. The first draft of the manuscript was written by WG and revised by RH and DL. Resources, supervision, and funding acquisition were performed by DL. All authors read and approved the final manuscript.

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Correspondence to Wei Guo.

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Guo, W., He, R., Zhao, Y. et al. Imbalanced metabolism induced NH4+ accumulation and its effect on the central metabolism of Methylomonas sp. ZR1. Int Microbiol 27, 49–66 (2024). https://doi.org/10.1007/s10123-023-00457-8

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