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Deep-sea microorganisms are critical in the ocean carbon cycle, and marine heterotrophic microorganisms secrete extracellular carbonic anhydrase (CA) to fix inorganic carbon, an important process in climate regulation. Extracellular CA provides a green method for fixing carbon dioxide into stable minerals containing Ca2+. However, studies on extracellular CA in deep-sea microorganisms are limited. In this study, Bacillus safensis yw6 was isolated from Mariana Trench sediments and three candidate extracellular CA genes (\u03b2-ca1, \u03b2-ca2, and \u03b3-ca) were identified by whole genome sequencing. Bioinformatics analyses showed that these CAs have different structural compositions, with the \u03b2-CA having \u03b1-helix and random coiling, whereas the \u03b3-CA has more random coiling and stretched strands. Heterologous expression in E. coli BL21 (DE3) showed that \u03b2-CA2 had the highest enzyme activity, followed by \u03b3-CA and \u03b2-CA1. Field emission scanning electron microscopy (FESEM) observations showed that the engineered strains with \u03b2-ca2 genes produced deposits that were like those from natural sources. This finding not only provides new perspectives for the utilization of deep-sea microbial resources, but also provides an important scientific basis for the molecular mechanisms of extracellular CAs of deep-sea microbes.<\/jats:p>","DOI":"10.3390\/molecules29245911","type":"journal-article","created":{"date-parts":[[2024,12,16]],"date-time":"2024-12-16T12:09:28Z","timestamp":1734350968000},"page":"5911","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in Bacillus safensis\u00a0yw6 from Mariana Trench"],"prefix":"10.3390","volume":"29","author":[{"given":"Xinyu","family":"Wang","sequence":"first","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pengna","family":"Wang","sequence":"additional","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-0091-6591","authenticated-orcid":false,"given":"Hancheng","family":"Zhao","sequence":"additional","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingying","family":"He","sequence":"additional","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Changfeng","family":"Qu","sequence":"additional","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jinlai","family":"Miao","sequence":"additional","affiliation":[{"name":"Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, China"},{"name":"Laboratory for Marine Drugs and Bioproducts, Qingdao Marine Science and Technology Center, Qingdao 266237, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Mwenketishi, G.T., Benkreira, H., and Rahmanian, N. (2023). A Comprehensive Review on Carbon Dioxide Sequestration Methods. Energies, 16.","DOI":"10.20944\/preprints202310.0127.v1"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1395","DOI":"10.1134\/S1064229322100064","article-title":"Research progress on microbial carbon sequestration in soil: A review","volume":"55","author":"Jiang","year":"2022","journal-title":"Eurasian Soil Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"825632","DOI":"10.3389\/fmicb.2022.825632","article-title":"Microbial communities in volcanic glacier ecosystems","volume":"13","author":"Cid","year":"2022","journal-title":"Front. Microbiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1016\/j.ijhydene.2023.05.307","article-title":"Role of microbial carbon capture cells in carbon sequestration and energy generation during wastewater treatment: A sustainable solution for cleaner environment","volume":"52","author":"Yaashikaa","year":"2024","journal-title":"Int. J. Hydrogen Energy"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1038\/s43017-022-00381-x","article-title":"Trends and variability in the ocean carbon sink","volume":"4","author":"Gruber","year":"2023","journal-title":"Nat. Rev. Earth Environ."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1016\/S1369-5274(98)80032-5","article-title":"Barophiles: Deep-sea microorganisms adapted to an extreme environment","volume":"1","author":"Horikoshi","year":"1999","journal-title":"Curr. Opin. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Gao, B., Jin, M., Li, L., Qu, W., and Zeng, R. (2017). Genome sequencing reveals the complex polysaccharide-degrading ability of novel deep-sea bacterium Flammeovirga pacifica WPAGA1. Front. Microbiol., 8.","DOI":"10.3389\/fmicb.2017.00600"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1907833","DOI":"10.1002\/adma.201907833","article-title":"Microbe-mediated extracellular and intracellular mineralization: Environmental, industrial, and biotechnological applications","volume":"32","author":"Qin","year":"2020","journal-title":"Adv. Mater."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"101045","DOI":"10.1016\/j.margen.2023.101045","article-title":"Metagenome-based analysis of carbon-fixing microorganisms and their carbon-fixing pathways in deep-sea sediments of the southwestern Indian Ocean","volume":"70","author":"Yue","year":"2023","journal-title":"Mar. Genom."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"271","DOI":"10.1016\/j.procbio.2019.12.018","article-title":"Influencing factors and formation mechanism of CaCO3 precipitation induced by microbial carbonic anhydrase","volume":"91","author":"Zheng","year":"2020","journal-title":"Process Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"56","DOI":"10.3390\/metabo7040056","article-title":"An Overview of the Bacterial Carbonic Anhydrases","volume":"7","author":"Supuran","year":"2017","journal-title":"Metabolites"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1111\/j.1749-6632.1984.tb12315.x","article-title":"Kinetics and Mechanism of Carbonic Anhydrase Isoenzymes","volume":"429","author":"Lindskog","year":"1984","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"15184","DOI":"10.1073\/pnas.96.26.15184","article-title":"Carbonic anhydrase is an ancient enzyme widespread in prokaryotes","volume":"96","author":"Smith","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1911","DOI":"10.3390\/jmse11101911","article-title":"Diversity of CO2 Concentrating Mechanisms in Macroalgae Photosynthesis: A Case Study of Ulva sp.","volume":"11","author":"Sun","year":"2023","journal-title":"J. Mar. Sci. Eng."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"655115","DOI":"10.3389\/fmolb.2021.655115","article-title":"Carbonic anhydrase in Pacific Abalone Haliotis discus hannai: Characterization, expression, and role in Biomineralization","volume":"8","author":"Sharker","year":"2021","journal-title":"Front. Mol. Biosci."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Anindita, H., and Meera, Y. (2023). Biomineralization of carbon dioxide by carbonic anhydrase. Biocatal. Agric. Biotechnol., 51.","DOI":"10.1016\/j.bcab.2023.102755"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"13570","DOI":"10.1039\/D2NR03199J","article-title":"Enhanced precipitation of magnesium carbonates using carbonic anhydrase","volume":"14","author":"Caulfield","year":"2022","journal-title":"Nanoscale"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1016\/j.procbio.2022.10.019","article-title":"Potential application of the immobilization of carbonic anhydrase based on metal organic framework supports","volume":"122","author":"Yuan","year":"2022","journal-title":"Process Biochem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"101475","DOI":"10.1016\/j.jcou.2021.101475","article-title":"Immobilization of carbonic anhydrase for CO2 capture and its industrial implementation: A review","volume":"47","author":"Luis","year":"2021","journal-title":"J. CO2 Util."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2613209","DOI":"10.1155\/2023\/2613209","article-title":"Bacteria-induced calcite precipitation for engineering and environmental applications","volume":"2023","author":"Baidya","year":"2023","journal-title":"Adv. Mater. Sci. Eng."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.jcou.2016.11.013","article-title":"Plasmonic nanostructured Zn\/ZnO composite enhances carbonic anhydrase driven photocatalytic hydrogen generation","volume":"17","author":"Mangrulkar","year":"2016","journal-title":"J. CO2 Util."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1042\/BCJ20160115","article-title":"Structure and function of carbonic anhydrases","volume":"473","author":"Supuran","year":"2016","journal-title":"Biochem. J."},{"key":"ref_23","first-page":"52","article-title":"Carbonic anhydrase II deficiency","volume":"294","author":"Whyte","year":"2023","journal-title":"Bone"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"19","DOI":"10.3390\/metabo8010019","article-title":"Carbonic anhydrases: Role in pH control and cancer","volume":"8","author":"Mboge","year":"2018","journal-title":"Metabolites"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"N\u00f6lke, G., Barsoum, M., Houdelet, M., Arcal\u00eds, E., Kreuzaler, F., Fischer, R., and Schillberg, S. (2019). The integration of algal carbon concentration mechanism components into tobacco chloroplasts increases photosynthetic efficiency and biomass. Biotechnol. J., 14.","DOI":"10.1002\/biot.201800170"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2827","DOI":"10.1111\/nph.17087","article-title":"High CO2-and pathogen-driven expression of the carbonic anhydrase \u03b2CA3 confers basal immunity in tomato","volume":"229","author":"Hu","year":"2020","journal-title":"N. Phytol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1186\/1756-3305-7-403","article-title":"Beta carbonic anhydrases: Novel targets for pesticides and anti-parasitic agents in agriculture and livestock husbandry","volume":"7","author":"Barker","year":"2014","journal-title":"Parasites Vectors"},{"key":"ref_28","first-page":"51","article-title":"Effect of grinding method on particle morphology and flotation behavior of calcite","volume":"40","author":"Jiangfeng","year":"2020","journal-title":"Conserv. Util. Miner. Resour."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1007\/s11430-015-5202-7","article-title":"The research of typical microbial functional group reveals a new oceanic carbon sequestration mechanism\u2212A case of innovative method promoting scientific discovery","volume":"59","author":"Zhang","year":"2016","journal-title":"Sci. China Earth Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"9679","DOI":"10.1073\/pnas.1918114117","article-title":"Metrics that matter for assessing the ocean biological carbon pump","volume":"117","author":"Buesseler","year":"2020","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"104359","DOI":"10.1016\/j.earscirev.2023.104359","article-title":"Quantitative and mechanistic understanding of the open ocean carbonate pump-perspectives for remote sensing and autonomous in situ observation","volume":"239","author":"Neukermans","year":"2023","journal-title":"Earth-Sci. Rev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"116685","DOI":"10.1016\/j.marpolbul.2024.116685","article-title":"Enhancing CO2 storage and marine carbon sink based on seawater mineral carbonation","volume":"206","author":"Liu","year":"2024","journal-title":"Mar. Pollut. Bull."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"173","DOI":"10.1016\/j.gene.2016.05.024","article-title":"Identification and expression of a novel carbonic anhydrase isozyme in the pufferfish Takifugu vermicularis","volume":"588","author":"Sumi","year":"2016","journal-title":"Gene"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"7413","DOI":"10.3390\/ijms22147413","article-title":"The Role of Extracellular Carbonic Anhydrase in Biogeochemical Cycling: Recent Advances and Climate Change Responses","volume":"22","author":"Latif","year":"2021","journal-title":"Int. J. Mol. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s00203-021-02677-w","article-title":"Microbial CO2 fixation and biotechnology in reducing industrial CO2 emissions","volume":"204","author":"Kajla","year":"2022","journal-title":"Arch. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Singh, S., Lomelino, C.L., Mboge, M.Y., Frost, S.C., and McKenna, R. (2018). Cancer Drug Development of Carbonic Anhydrase Inhibitors beyond the Active Site. Molecules, 23.","DOI":"10.3390\/molecules23051045"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"76","DOI":"10.3390\/microorganisms10010076","article-title":"New biocalcifying marine bacterial strains isolated from calcareous deposits and immediate surroundings","volume":"10","author":"Vincent","year":"2021","journal-title":"Microorganisms"}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/24\/5911\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:55:22Z","timestamp":1760115322000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/29\/24\/5911"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,14]]},"references-count":37,"journal-issue":{"issue":"24","published-online":{"date-parts":[[2024,12]]}},"alternative-id":["molecules29245911"],"URL":"https:\/\/doi.org\/10.3390\/molecules29245911","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,12,14]]}}}