Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade
Ursodeoxycholic acid (UDCA) can be used as a drug to treat various liver and bile diseases. Currently, the biological synthesis of UDCA is predominantly conducted via a two-step enzymatic process in which synthesis is catalyzed by 7α-hydroxysteroid dehydrogenase (7α-HSDH) and 7β-hydroxysteroid dehyd...
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| Format: | Article |
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1538237/full |
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| author | Xue Zhang Xue Zhang Jiagang Xin Mengyu Liu Mengyu Liu Yue Zhang Yue Zhang Haoni Luan Haoni Luan Wei Feng Wei Feng Fei Wang Fei Wang Wei Xu Wei Xu Peng Song Peng Song |
| author_facet | Xue Zhang Xue Zhang Jiagang Xin Mengyu Liu Mengyu Liu Yue Zhang Yue Zhang Haoni Luan Haoni Luan Wei Feng Wei Feng Fei Wang Fei Wang Wei Xu Wei Xu Peng Song Peng Song |
| author_sort | Xue Zhang |
| collection | DOAJ |
| description | Ursodeoxycholic acid (UDCA) can be used as a drug to treat various liver and bile diseases. Currently, the biological synthesis of UDCA is predominantly conducted via a two-step enzymatic process in which synthesis is catalyzed by 7α-hydroxysteroid dehydrogenase (7α-HSDH) and 7β-hydroxysteroid dehydrogenase (7β-HSDH) in succession, utilizing chenodeoxycholic acid (CDCA) as the substrate. In this study, an engineered Escherichia coli (E. coli) strain, designated UCA23, was constructed. This strain coexpressed four enzymes under the control of three independent T7 promoters: lactate dehydrogenase (LDH) derived from Lactobacillus delbrueckii, glucose dehydrogenase (GDH) derived from Priestia megaterium, 7α-HSDH derived from E. coli, and 7β-HSDH derived from Ruminococcus torques, enabling the whole-cell catalytic synthesis of UDCA from CDCA. This study systematically optimized the reaction parameters, including temperature, pH, and the addition of organic solvents and surfactants, for the whole-cell catalytic synthesis of UDCA by UCA23, and at the 2 L level, a UDCA conversion rate of 99% was achieved with 100 mM CDCA in 2 h, which is the highest level of conversion of a high-concentration CDCA substrate reported to date. |
| format | Article |
| id | doaj-art-118268246a6a487d871f030d6c7c5c7e |
| institution | Kabale University |
| issn | 1664-302X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj-art-118268246a6a487d871f030d6c7c5c7e2025-01-27T13:29:02ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011610.3389/fmicb.2025.15382371538237Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascadeXue Zhang0Xue Zhang1Jiagang Xin2Mengyu Liu3Mengyu Liu4Yue Zhang5Yue Zhang6Haoni Luan7Haoni Luan8Wei Feng9Wei Feng10Fei Wang11Fei Wang12Wei Xu13Wei Xu14Peng Song15Peng Song16School of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaShandong Aobo Biotech Co. Ltd., Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaSchool of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, ChinaNational Key Laboratory of Macromolecular Drug Development and Manufacturing, Liaocheng, ChinaUrsodeoxycholic acid (UDCA) can be used as a drug to treat various liver and bile diseases. Currently, the biological synthesis of UDCA is predominantly conducted via a two-step enzymatic process in which synthesis is catalyzed by 7α-hydroxysteroid dehydrogenase (7α-HSDH) and 7β-hydroxysteroid dehydrogenase (7β-HSDH) in succession, utilizing chenodeoxycholic acid (CDCA) as the substrate. In this study, an engineered Escherichia coli (E. coli) strain, designated UCA23, was constructed. This strain coexpressed four enzymes under the control of three independent T7 promoters: lactate dehydrogenase (LDH) derived from Lactobacillus delbrueckii, glucose dehydrogenase (GDH) derived from Priestia megaterium, 7α-HSDH derived from E. coli, and 7β-HSDH derived from Ruminococcus torques, enabling the whole-cell catalytic synthesis of UDCA from CDCA. This study systematically optimized the reaction parameters, including temperature, pH, and the addition of organic solvents and surfactants, for the whole-cell catalytic synthesis of UDCA by UCA23, and at the 2 L level, a UDCA conversion rate of 99% was achieved with 100 mM CDCA in 2 h, which is the highest level of conversion of a high-concentration CDCA substrate reported to date.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1538237/fullchenodeoxycholic acidursodeoxycholic acidwhole-cell catalysis7α-hydroxysteroid dehydrogenase7β-hydroxysteroid dehydrogenaselactate dehydrogenase |
| spellingShingle | Xue Zhang Xue Zhang Jiagang Xin Mengyu Liu Mengyu Liu Yue Zhang Yue Zhang Haoni Luan Haoni Luan Wei Feng Wei Feng Fei Wang Fei Wang Wei Xu Wei Xu Peng Song Peng Song Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade Frontiers in Microbiology chenodeoxycholic acid ursodeoxycholic acid whole-cell catalysis 7α-hydroxysteroid dehydrogenase 7β-hydroxysteroid dehydrogenase lactate dehydrogenase |
| title | Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade |
| title_full | Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade |
| title_fullStr | Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade |
| title_full_unstemmed | Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade |
| title_short | Cost-effective whole-cell biosynthesis of ursodeoxycholic acid using engineered Escherichia coli with a multienzyme cascade |
| title_sort | cost effective whole cell biosynthesis of ursodeoxycholic acid using engineered escherichia coli with a multienzyme cascade |
| topic | chenodeoxycholic acid ursodeoxycholic acid whole-cell catalysis 7α-hydroxysteroid dehydrogenase 7β-hydroxysteroid dehydrogenase lactate dehydrogenase |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2025.1538237/full |
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