Biochemical characterization of a psychrophilic and halotolerant α–carbonic anhydrase from a deep-sea bacterium, <i>Photobacterium profundum</i>

Vijayakumar Somalinga; Emily Foss; Amy M. Grunden; Vijayakumar Somalinga; Emily Foss; Amy M. Grunden

doi:10.3934/microbiol.2023028

AIMS Microbiology

2023, Volume 9, Issue 3: 540-553. doi: 10.3934/microbiol.2023028

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Research article

Biochemical characterization of a psychrophilic and halotolerant α–carbonic anhydrase from a deep-sea bacterium, Photobacterium profundum

1.
Department of Biological & Biomedical Sciences, Southwestern Oklahoma State University, 100 Campus Drive, Weatherford, OK 73096, USA
2.
Department of Plant and Microbial Biology, North Carolina State University, 4550A Thomas Hall, Campus Box 7612, Raleigh, NC 27695, USA

Received: 03 February 2023 Revised: 20 May 2023 Accepted: 24 May 2023 Published: 19 June 2023

Prokaryotic α–carbonic anhydrases (α-CA) are metalloenzymes that catalyze the reversible hydration of CO₂ to bicarbonate and proton. We had reported the first crystal structure of a pyschrohalophilic α–CA from a deep-sea bacterium, Photobacterium profundum SS9. In this manuscript, we report the first biochemical characterization of P. profundum α–CA (PprCA) which revealed several catalytic properties that are atypical for this class of CA's. Purified PprCA exhibited maximal catalytic activity at psychrophilic temperatures with substantial decrease in activity at mesophilic and thermophilic range. Similar to other α–CA's, Ppr9A showed peak activity at alkaline pH (pH 11), although, PprCA retained 88% of its activity even at acidic pH (pH 5). Exposing PprCA to varying concentrations of oxidizing and reducing agents revealed that N-terminal cysteine residues in PprCA may play a role in the structural stability of the enzyme. Although inefficient in CO₂ hydration activity under mesophilic and thermophilic temperatures, PprCA exhibited salt-dependent thermotolerance and catalytic activity under extreme halophilic conditions. Similar to other well-characterized α–CA's, PprCA is also inhibited by monovalent anions even at low concentrations. Finally, we demonstrate that PprCA accelerates CO₂ biomineralization to calcium carbonate under alkaline conditions.
- Photobacterium profundum,
- α-carbonic anhydrase,
- halotolerance,
- biomineralization,
- carbon capture
Citation: Vijayakumar Somalinga, Emily Foss, Amy M. Grunden. Biochemical characterization of a psychrophilic and halotolerant α–carbonic anhydrase from a deep-sea bacterium, Photobacterium profundum[J]. AIMS Microbiology, 2023, 9(3): 540-553. doi: 10.3934/microbiol.2023028

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Abstract

Prokaryotic α–carbonic anhydrases (α-CA) are metalloenzymes that catalyze the reversible hydration of CO₂ to bicarbonate and proton. We had reported the first crystal structure of a pyschrohalophilic α–CA from a deep-sea bacterium, Photobacterium profundum SS9. In this manuscript, we report the first biochemical characterization of P. profundum α–CA (PprCA) which revealed several catalytic properties that are atypical for this class of CA's. Purified PprCA exhibited maximal catalytic activity at psychrophilic temperatures with substantial decrease in activity at mesophilic and thermophilic range. Similar to other α–CA's, Ppr9A showed peak activity at alkaline pH (pH 11), although, PprCA retained 88% of its activity even at acidic pH (pH 5). Exposing PprCA to varying concentrations of oxidizing and reducing agents revealed that N-terminal cysteine residues in PprCA may play a role in the structural stability of the enzyme. Although inefficient in CO₂ hydration activity under mesophilic and thermophilic temperatures, PprCA exhibited salt-dependent thermotolerance and catalytic activity under extreme halophilic conditions. Similar to other well-characterized α–CA's, PprCA is also inhibited by monovalent anions even at low concentrations. Finally, we demonstrate that PprCA accelerates CO₂ biomineralization to calcium carbonate under alkaline conditions.

Acknowledgments

A part of this work was supported by NSF OK-EPSCoR REU program under the grant number OIA-1946093 awarded to Vijay Somalinga and Emily Foss. This work was also supported by the SWOSU Guy Hagin Endowment fund awarded to Vijay Somalinga, and by the National Science Foundation EFRI program under grant number EFRI 1332341 awarded to Amy Grunden.

Conflict of interest

The authors declare no competing interests.

Use of AI tools declaration

The authors declare that they have not used Artificial Intelligence (AI) tools in the creation of this article.

Author contributions

VS & AMG designed the study and the experiments. VS and AMG supervised the project. VS and EF performed the experiments. VS wrote the manuscript. VS, EF and AMG edited, and reviewed the manuscript. All authors approved this final submission.

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