Osmotic stress as a factor for regulating <i>E. coli</i> hydrogenase activity and enhancing H<sub>2</sub> production during mixed carbon sources fermentation

Anush Babayan; Anait Vassilian; Karen Trchounian; Anush Babayan; Anait Vassilian; Karen Trchounian

doi:10.3934/microbiol.2023037

AIMS Microbiology

2023, Volume 9, Issue 4: 724-737. doi: 10.3934/microbiol.2023037

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Research article

Osmotic stress as a factor for regulating E. coli hydrogenase activity and enhancing H₂ production during mixed carbon sources fermentation

1.
Department of Biochemistry, Microbiology and Biotechnology, Faculty of Biology, Yerevan State University, 0025 Yerevan, Armenia
2.
Scientific-Research Institute of Biology, Faculty of Biology, Yerevan State University, 0025 Yerevan, Armenia
3.
Microbial Biotechnologies and Biofuel Innovation Center, Yerevan State University, 0025 Yerevan, Armenia
4.
Department of Ecology and Nature Protection, Faculty of Biology, Yerevan State University, 0025 Yerevan, Armenia

Received: 22 July 2023 Revised: 18 October 2023 Accepted: 27 October 2023 Published: 06 November 2023

Escherichia coli performs mixed-acid fermentation and produces molecular hydrogen (H₂) via reversible hydrogenases (Hyd). H₂ producing activity was investigated during hyper- and hypo-osmotic stress conditions when a mixture of carbon sources (glucose and glycerol) was fermented at different pHs. Hyper-osmotic stress decreased H₂ production rate (V_H2) ~30 % in wild type at pH 7.5 when glucose was supplemented, while addition of formate stimulated V_H2 ~45% compared to hypo-stress conditions. Only in hyfG in formate assays was V_H2 inhibited ~25% compared to hypo-stress conditions. In hypo-stress conditions addition of glycerol increased V_H2 ~2 and 3 fold in hybC and hyfG mutants, respectively, compared to wild type. At pH 6.5 hyper-osmotic stress stimulated V_H2 ~2 fold in all strains except hyaB mutant when glucose was supplemented, while in formate assays significant stimulation (~3 fold) was determined in hybC mutant. At pH 5.5 hyper-osmotic stress inhibited V_H2 ~30% in wild type when glucose was supplemented, but in formate assays it was stimulated in all strains except hyfG. Taken together, it can be concluded that, depending on external pH and absence of Hyd enzymes in stationary-phase-grown osmotically stressed E. coli cells, H₂ production can be stimulated significantly which can be applied in developing H₂ production biotechnology.
- Escherichia coli,
- mixed carbon fermentation,
- Hyd enzymes,
- osmotic stress,
- pH
Citation: Anush Babayan, Anait Vassilian, Karen Trchounian. Osmotic stress as a factor for regulating E. coli hydrogenase activity and enhancing H2 production during mixed carbon sources fermentation[J]. AIMS Microbiology, 2023, 9(4): 724-737. doi: 10.3934/microbiol.2023037

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Abstract

Escherichia coli performs mixed-acid fermentation and produces molecular hydrogen (H₂) via reversible hydrogenases (Hyd). H₂ producing activity was investigated during hyper- and hypo-osmotic stress conditions when a mixture of carbon sources (glucose and glycerol) was fermented at different pHs. Hyper-osmotic stress decreased H₂ production rate (V_H2) ~30 % in wild type at pH 7.5 when glucose was supplemented, while addition of formate stimulated V_H2 ~45% compared to hypo-stress conditions. Only in hyfG in formate assays was V_H2 inhibited ~25% compared to hypo-stress conditions. In hypo-stress conditions addition of glycerol increased V_H2 ~2 and 3 fold in hybC and hyfG mutants, respectively, compared to wild type. At pH 6.5 hyper-osmotic stress stimulated V_H2 ~2 fold in all strains except hyaB mutant when glucose was supplemented, while in formate assays significant stimulation (~3 fold) was determined in hybC mutant. At pH 5.5 hyper-osmotic stress inhibited V_H2 ~30% in wild type when glucose was supplemented, but in formate assays it was stimulated in all strains except hyfG. Taken together, it can be concluded that, depending on external pH and absence of Hyd enzymes in stationary-phase-grown osmotically stressed E. coli cells, H₂ production can be stimulated significantly which can be applied in developing H₂ production biotechnology.

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Availability of data and materials

The datasets used and/or analyzed for the current study are available from the corresponding author on reasonable request.

Competing interests

The authors declare that they have no competing interests.

Funding

The study was supported by a Basic Research Support Grant to KT and a Research grant from the Science Committee, Ministry of Education, Science, Culture and Sports of Armenia, 21AG-1F043.

Authors' contributions

AB conducted the experiments. AV and KT designed the experiments and analyzed the data. KT drafted and wrote the manuscript and finalized the submission.

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