A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application
The low productivity in long fermentation duration and high-salt working conditions limit the application of L-glutaminase in soy sauce brewing. In this study, a novel L-glutaminase (LreuglsA) with eminent salt tolerance was mined and achieved more than 70% activity with 30% NaCl. To improve the rob...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/9/444 |
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| author | Hengwei Zhang Mengkai Hu Qing Wang Fei Liu Meijuan Xu Xian Zhang Zhiming Rao |
| author_facet | Hengwei Zhang Mengkai Hu Qing Wang Fei Liu Meijuan Xu Xian Zhang Zhiming Rao |
| author_sort | Hengwei Zhang |
| collection | DOAJ |
| description | The low productivity in long fermentation duration and high-salt working conditions limit the application of L-glutaminase in soy sauce brewing. In this study, a novel L-glutaminase (LreuglsA) with eminent salt tolerance was mined and achieved more than 70% activity with 30% NaCl. To improve the robustness of the enzyme at different fermentation strategies, mutation LreuglsA<sup>H105K</sup> was built by a computer-aided design, and the recombinant protein expression level, an essential parameter in industrial applications, was increased 5.61-fold with the synthetic biology strategy by improving the mRNA stability. Finally, the LreuglsA<sup>H105K</sup> functional expression box was contributed to <i>Bacillus subtilis</i> 168 by auxotrophic complementation, and the production in a 5-L bioreactor was improved to 2516.78 ± 20.83 U mL<sup>−1</sup>, the highest production ever reported. When the immobilized cells were applied to high-salt dilute-state soy sauce brewing, the L-glutamate level was increased by 45.9%. This work provides insight into the salt-tolerant enzyme for improving the efficiency of industrial applications. |
| first_indexed | 2024-03-10T00:04:05Z |
| format | Article |
| id | doaj.art-31310524262f4348b9c02b42083528e7 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-10T00:04:05Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-31310524262f4348b9c02b42083528e72023-11-23T16:10:47ZengMDPI AGFermentation2311-56372022-09-018944410.3390/fermentation8090444A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and ApplicationHengwei Zhang0Mengkai Hu1Qing Wang2Fei Liu3Meijuan Xu4Xian Zhang5Zhiming Rao6Key Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Industrial Biotechnology of the Ministry of Education, Laboratory of Applied Microorganisms and Metabolic Engineering, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaThe low productivity in long fermentation duration and high-salt working conditions limit the application of L-glutaminase in soy sauce brewing. In this study, a novel L-glutaminase (LreuglsA) with eminent salt tolerance was mined and achieved more than 70% activity with 30% NaCl. To improve the robustness of the enzyme at different fermentation strategies, mutation LreuglsA<sup>H105K</sup> was built by a computer-aided design, and the recombinant protein expression level, an essential parameter in industrial applications, was increased 5.61-fold with the synthetic biology strategy by improving the mRNA stability. Finally, the LreuglsA<sup>H105K</sup> functional expression box was contributed to <i>Bacillus subtilis</i> 168 by auxotrophic complementation, and the production in a 5-L bioreactor was improved to 2516.78 ± 20.83 U mL<sup>−1</sup>, the highest production ever reported. When the immobilized cells were applied to high-salt dilute-state soy sauce brewing, the L-glutamate level was increased by 45.9%. This work provides insight into the salt-tolerant enzyme for improving the efficiency of industrial applications.https://www.mdpi.com/2311-5637/8/9/444L-glutaminasecomputer-aided designexpression level5-L bioreactorhigh-salt dilute-state soy sauce |
| spellingShingle | Hengwei Zhang Mengkai Hu Qing Wang Fei Liu Meijuan Xu Xian Zhang Zhiming Rao A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application Fermentation L-glutaminase computer-aided design expression level 5-L bioreactor high-salt dilute-state soy sauce |
| title | A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application |
| title_full | A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application |
| title_fullStr | A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application |
| title_full_unstemmed | A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application |
| title_short | A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application |
| title_sort | novel salt tolerant l glutaminase efficient functional expression computer aided design and application |
| topic | L-glutaminase computer-aided design expression level 5-L bioreactor high-salt dilute-state soy sauce |
| url | https://www.mdpi.com/2311-5637/8/9/444 |
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