Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis
Guanidinoacetate (GAA) is a naturally occurring amino acid derivative and the direct precursor of creatine, which is widely used in feed additives and the pharmaceutical industry. The current industrial synthesis of GAA is based on chemical methods, which limits the application of GAA. Here, a biolo...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/3/116 |
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| author | Kun Yan Rongzhen Tian Linpei Zhang Xueqin Lv Long Liu Yanfeng Liu |
| author_facet | Kun Yan Rongzhen Tian Linpei Zhang Xueqin Lv Long Liu Yanfeng Liu |
| author_sort | Kun Yan |
| collection | DOAJ |
| description | Guanidinoacetate (GAA) is a naturally occurring amino acid derivative and the direct precursor of creatine, which is widely used in feed additives and the pharmaceutical industry. The current industrial synthesis of GAA is based on chemical methods, which limits the application of GAA. Here, a biological approach is developed for food safety GAA production via whole-cell biocatalysis by the generally regarded as safe (GRAS) bacterium <i>Bacillus subtilis</i>. First, we introduced a heterologous arginine: glycine amidinotransferase (AgaT) from <i>Amycolatopsis kentuckyensis</i> into <i>B. subtilis</i> and optimized its expression level using strategies including: promoter optimization, ribosome binding site (RBS) and N-terminal coding sequence (NCS) screening. In order to alleviate the waste of arginine and the inhibition of AgaT by ornithine, we optimized the natural ornithine cycle in <i>B. subtilis</i>. At the same time, the first gene in the glycine degradation pathway was knocked out. After optimization using these strategies, the titer of GAA was 4.26 g/L with a productivity of 0.21 g/L/h in 20 h, which provides a new method for the biosynthesis of GAA. |
| first_indexed | 2024-03-09T19:51:50Z |
| format | Article |
| id | doaj.art-0cfc62637dd343fab28c81d711707f22 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-09T19:51:50Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-0cfc62637dd343fab28c81d711707f222023-11-24T01:08:39ZengMDPI AGFermentation2311-56372022-03-018311610.3390/fermentation8030116Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell CatalysisKun Yan0Rongzhen Tian1Linpei Zhang2Xueqin Lv3Long Liu4Yanfeng Liu5Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaKey Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, ChinaGuanidinoacetate (GAA) is a naturally occurring amino acid derivative and the direct precursor of creatine, which is widely used in feed additives and the pharmaceutical industry. The current industrial synthesis of GAA is based on chemical methods, which limits the application of GAA. Here, a biological approach is developed for food safety GAA production via whole-cell biocatalysis by the generally regarded as safe (GRAS) bacterium <i>Bacillus subtilis</i>. First, we introduced a heterologous arginine: glycine amidinotransferase (AgaT) from <i>Amycolatopsis kentuckyensis</i> into <i>B. subtilis</i> and optimized its expression level using strategies including: promoter optimization, ribosome binding site (RBS) and N-terminal coding sequence (NCS) screening. In order to alleviate the waste of arginine and the inhibition of AgaT by ornithine, we optimized the natural ornithine cycle in <i>B. subtilis</i>. At the same time, the first gene in the glycine degradation pathway was knocked out. After optimization using these strategies, the titer of GAA was 4.26 g/L with a productivity of 0.21 g/L/h in 20 h, which provides a new method for the biosynthesis of GAA.https://www.mdpi.com/2311-5637/8/3/116<i>B. subtilis</i>guanidinoacetateornithine cyclewhole-cell catalysis |
| spellingShingle | Kun Yan Rongzhen Tian Linpei Zhang Xueqin Lv Long Liu Yanfeng Liu Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis Fermentation <i>B. subtilis</i> guanidinoacetate ornithine cycle whole-cell catalysis |
| title | Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis |
| title_full | Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis |
| title_fullStr | Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis |
| title_full_unstemmed | Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis |
| title_short | Biosynthesis of Guanidinoacetate by <i>Bacillus subtilis</i> Whole-Cell Catalysis |
| title_sort | biosynthesis of guanidinoacetate by i bacillus subtilis i whole cell catalysis |
| topic | <i>B. subtilis</i> guanidinoacetate ornithine cycle whole-cell catalysis |
| url | https://www.mdpi.com/2311-5637/8/3/116 |
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