Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench
The exploration and exploitation of deep-sea microbial resources is of great scientific value for understanding biological evolution under extreme conditions. Deep-sea microorganisms are critical in the ocean carbon cycle, and marine heterotrophic microorganisms secrete extracellular carbonic anhydr...
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2024-12-01
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| author | Xinyu Wang Pengna Wang Hancheng Zhao Yingying He Changfeng Qu Jinlai Miao |
| author_facet | Xinyu Wang Pengna Wang Hancheng Zhao Yingying He Changfeng Qu Jinlai Miao |
| author_sort | Xinyu Wang |
| collection | DOAJ |
| description | The exploration and exploitation of deep-sea microbial resources is of great scientific value for understanding biological evolution under extreme conditions. 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 Ca<sup>2+</sup>. However, studies on extracellular CA in deep-sea microorganisms are limited. In this study, <i>Bacillus safensis yw6</i> was isolated from Mariana Trench sediments and three candidate extracellular CA genes (<i>β-ca</i>1, <i>β-ca</i>2, and <i>γ-ca</i>) were identified by whole genome sequencing. Bioinformatics analyses showed that these CAs have different structural compositions, with the β-CA having α-helix and random coiling, whereas the γ-CA has more random coiling and stretched strands. Heterologous expression in E. coli BL21 (DE3) showed that β-CA2 had the highest enzyme activity, followed by γ-CA and β-CA1. Field emission scanning electron microscopy (FESEM) observations showed that the engineered strains with <i>β-ca</i>2 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. |
| format | Article |
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| issn | 1420-3049 |
| language | English |
| publishDate | 2024-12-01 |
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| spelling | doaj-art-9901a6419c8348948ea3ccf88b1fd7722025-08-20T02:01:20ZengMDPI AGMolecules1420-30492024-12-012924591110.3390/molecules29245911Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana TrenchXinyu Wang0Pengna Wang1Hancheng Zhao2Yingying He3Changfeng Qu4Jinlai Miao5Marine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaMarine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaMarine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaMarine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaMarine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaMarine Natural Products Research and Development Key Laboratory of Qingdao, First Institute of Oceanography, Ministry of Natural Resources, Qingdao 266061, ChinaThe exploration and exploitation of deep-sea microbial resources is of great scientific value for understanding biological evolution under extreme conditions. 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 Ca<sup>2+</sup>. However, studies on extracellular CA in deep-sea microorganisms are limited. In this study, <i>Bacillus safensis yw6</i> was isolated from Mariana Trench sediments and three candidate extracellular CA genes (<i>β-ca</i>1, <i>β-ca</i>2, and <i>γ-ca</i>) were identified by whole genome sequencing. Bioinformatics analyses showed that these CAs have different structural compositions, with the β-CA having α-helix and random coiling, whereas the γ-CA has more random coiling and stretched strands. Heterologous expression in E. coli BL21 (DE3) showed that β-CA2 had the highest enzyme activity, followed by γ-CA and β-CA1. Field emission scanning electron microscopy (FESEM) observations showed that the engineered strains with <i>β-ca</i>2 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.https://www.mdpi.com/1420-3049/29/24/5911extracellular carbonic anhydrase<i>Bacillus schafferii</i>bioinformatics analysisheterologous expression |
| spellingShingle | Xinyu Wang Pengna Wang Hancheng Zhao Yingying He Changfeng Qu Jinlai Miao Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench Molecules extracellular carbonic anhydrase <i>Bacillus schafferii</i> bioinformatics analysis heterologous expression |
| title | Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench |
| title_full | Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench |
| title_fullStr | Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench |
| title_full_unstemmed | Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench |
| title_short | Heterologous Expression and Functional Verification of Extracellular Carbonic Anhydrases in <i>Bacillus safensis yw6</i> from Mariana Trench |
| title_sort | heterologous expression and functional verification of extracellular carbonic anhydrases in i bacillus safensis yw6 i from mariana trench |
| topic | extracellular carbonic anhydrase <i>Bacillus schafferii</i> bioinformatics analysis heterologous expression |
| url | https://www.mdpi.com/1420-3049/29/24/5911 |
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