A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation
κ-carrageenases are members of the glycoside hydrolase family 16 (GH16) that hydrolyze sulfated galactans in red algae, known as κ-carrageenans. In this study, a novel κ-carrageenase gene from the marine bacterium <i>Rhodopirellula sallentina</i> SM41 (RsCgk) was discovered via the genom...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-12-01
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| Series: | Marine Drugs |
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| Online Access: | https://www.mdpi.com/1660-3397/20/12/783 |
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| author | Yong-Hui Zhang Yi-Ying Chen Xiao-Yan Zhuang Qiong Xiao Jun Chen Fu-Quan Chen Qiu-Ming Yang Hui-Fen Weng Bai-Shan Fang An-Feng Xiao |
| author_facet | Yong-Hui Zhang Yi-Ying Chen Xiao-Yan Zhuang Qiong Xiao Jun Chen Fu-Quan Chen Qiu-Ming Yang Hui-Fen Weng Bai-Shan Fang An-Feng Xiao |
| author_sort | Yong-Hui Zhang |
| collection | DOAJ |
| description | κ-carrageenases are members of the glycoside hydrolase family 16 (GH16) that hydrolyze sulfated galactans in red algae, known as κ-carrageenans. In this study, a novel κ-carrageenase gene from the marine bacterium <i>Rhodopirellula sallentina</i> SM41 (RsCgk) was discovered via the genome mining approach. There are currently no reports on κ-carrageenase from the <i>Rhodopirellula</i> genus, and RsCgk shares a low identity (less than 65%) with κ- carrageenase from other genera. The RsCgk was heterologously overexpressed in <i>Escherichia coli</i> BL21 and characterized for its enzymatic properties. RsCgk exhibited maximum activity at pH 7.0 and 40 °C, and 50% of its initial activity was retained after incubating at 30 °C for 2 h. More than 70% of its activity was maintained after incubation at pH 6.0–8.0 and 4 °C for 24 h. As a marine derived enzyme, RsCgk showed excellent salt tolerance, retaining full activity in 1.2 M NaCl, and the addition of NaCl greatly enhanced its thermal stability. Mass spectrometry analysis of the RsCgk hydrolysis products revealed that the enzyme had high degradation specificity and mainly produced κ-carrageenan disaccharide. Comparative molecular dynamics simulations revealed that the conformational changes of tunnel-forming loops under salt environments may cause the deactivation or stabilization of RsCgk. Our results demonstrated that RsCgk could be utilized as a potential tool enzyme for efficient production of κ-carrageenan oligosaccharides under high salt conditions. |
| first_indexed | 2024-03-09T16:10:08Z |
| format | Article |
| id | doaj.art-88edb9fb151d481b9b869ab6ab19bfff |
| institution | Directory Open Access Journal |
| issn | 1660-3397 |
| language | English |
| last_indexed | 2024-03-09T16:10:08Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Marine Drugs |
| spelling | doaj.art-88edb9fb151d481b9b869ab6ab19bfff2023-11-24T16:20:07ZengMDPI AGMarine Drugs1660-33972022-12-01201278310.3390/md20120783A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism InvestigationYong-Hui Zhang0Yi-Ying Chen1Xiao-Yan Zhuang2Qiong Xiao3Jun Chen4Fu-Quan Chen5Qiu-Ming Yang6Hui-Fen Weng7Bai-Shan Fang8An-Feng Xiao9College of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, ChinaCollege of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361021, ChinaCollege of Food and Biological Engineering, Jimei University, Xiamen 361021, Chinaκ-carrageenases are members of the glycoside hydrolase family 16 (GH16) that hydrolyze sulfated galactans in red algae, known as κ-carrageenans. In this study, a novel κ-carrageenase gene from the marine bacterium <i>Rhodopirellula sallentina</i> SM41 (RsCgk) was discovered via the genome mining approach. There are currently no reports on κ-carrageenase from the <i>Rhodopirellula</i> genus, and RsCgk shares a low identity (less than 65%) with κ- carrageenase from other genera. The RsCgk was heterologously overexpressed in <i>Escherichia coli</i> BL21 and characterized for its enzymatic properties. RsCgk exhibited maximum activity at pH 7.0 and 40 °C, and 50% of its initial activity was retained after incubating at 30 °C for 2 h. More than 70% of its activity was maintained after incubation at pH 6.0–8.0 and 4 °C for 24 h. As a marine derived enzyme, RsCgk showed excellent salt tolerance, retaining full activity in 1.2 M NaCl, and the addition of NaCl greatly enhanced its thermal stability. Mass spectrometry analysis of the RsCgk hydrolysis products revealed that the enzyme had high degradation specificity and mainly produced κ-carrageenan disaccharide. Comparative molecular dynamics simulations revealed that the conformational changes of tunnel-forming loops under salt environments may cause the deactivation or stabilization of RsCgk. Our results demonstrated that RsCgk could be utilized as a potential tool enzyme for efficient production of κ-carrageenan oligosaccharides under high salt conditions.https://www.mdpi.com/1660-3397/20/12/783κ-carrageenaseκ-carrageenanheterologous expressionsalt-tolerance |
| spellingShingle | Yong-Hui Zhang Yi-Ying Chen Xiao-Yan Zhuang Qiong Xiao Jun Chen Fu-Quan Chen Qiu-Ming Yang Hui-Fen Weng Bai-Shan Fang An-Feng Xiao A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation Marine Drugs κ-carrageenase κ-carrageenan heterologous expression salt-tolerance |
| title | A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation |
| title_full | A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation |
| title_fullStr | A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation |
| title_full_unstemmed | A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation |
| title_short | A Novel κ-Carrageenase from Marine Bacterium <i>Rhodopirellula sallentina</i> SM41: Heterologous Expression, Biochemical Characterization and Salt-Tolerance Mechanism Investigation |
| title_sort | novel κ carrageenase from marine bacterium i rhodopirellula sallentina i sm41 heterologous expression biochemical characterization and salt tolerance mechanism investigation |
| topic | κ-carrageenase κ-carrageenan heterologous expression salt-tolerance |
| url | https://www.mdpi.com/1660-3397/20/12/783 |
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