Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i>
<span style="font-variant: small-caps;">l</span>-Cysteine is an important sulfur-containing amino acid with numerous applications in the pharmaceutical and cosmetic industries. The microbial production of <span style="font-variant: small-caps;">l</span>-cy...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-06-01
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| Series: | Fermentation |
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| Online Access: | https://www.mdpi.com/2311-5637/8/7/299 |
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| author | Xiaomei Zhang Zhenhang Sun Jinyu Bian Yujie Gao Dong Zhang Guoqiang Xu Xiaojuan Zhang Hui Li Jinsong Shi Zhenghong Xu |
| author_facet | Xiaomei Zhang Zhenhang Sun Jinyu Bian Yujie Gao Dong Zhang Guoqiang Xu Xiaojuan Zhang Hui Li Jinsong Shi Zhenghong Xu |
| author_sort | Xiaomei Zhang |
| collection | DOAJ |
| description | <span style="font-variant: small-caps;">l</span>-Cysteine is an important sulfur-containing amino acid with numerous applications in the pharmaceutical and cosmetic industries. The microbial production of <span style="font-variant: small-caps;">l</span>-cysteine has received substantial attention, and the supply of the precursor <span style="font-variant: small-caps;">l</span>-serine is important in <span style="font-variant: small-caps;">l</span>-cysteine biosynthesis. In this study, to achieve <span style="font-variant: small-caps;">l</span>-cysteine overproduction, we first increased <span style="font-variant: small-caps;">l</span>-serine production by deleting genes involved in the pathway of <span style="font-variant: small-caps;">l</span>-serine degradation to glycine (serine hydroxymethyl transferase, SHMT, encoded by <i>glyA</i> genes) in strain 4W (with <span style="font-variant: small-caps;">l</span>-serine titer of 1.1 g/L), thus resulting in strain 4WG with <span style="font-variant: small-caps;">l</span>-serine titer of 2.01 g/L. Second, the serine-biosensor based on the transcriptional regulator NCgl0581 of <i>C. glutamicum</i> was constructed in <i>E. coli,</i> and the validity and sensitivity of the biosensor were demonstrated in <i>E. coli</i>. Then 4WG was further evolved through adaptive laboratory evolution (ALE) combined with serine-biosensor, thus yielding the strain 4WGX with 4.13 g/L <span style="font-variant: small-caps;">l</span>-serine production. Moreover, the whole genome of the evolved strain 4WGX was sequenced, and ten non-synonymous mutations were found in the genome of strain 4WGX compared with strain 4W. Finally, 4WGX was used as the starting strain, and deletion of the <span style="font-variant: small-caps;">l</span>-cysteine desulfhydrases (encoded by <i>tnaA</i>), overexpression of serine acetyltransferase (encoded by <i>cysE</i>) and the key enzyme of transport pathway (encoded by <i>ydeD</i>) were performed in strain 4WGX. The recombinant strain 4WGX-∆<i>tnaA</i>-<i>cysE</i>-<i>ydeD</i> can produce 313.4 mg/L of <span style="font-variant: small-caps;">l</span>-cysteine using glycerol as the carbon source. This work provides an efficient method for the biosynthesis of value-added commodity products associated with glycerol conversion. |
| first_indexed | 2024-03-09T03:26:15Z |
| format | Article |
| id | doaj.art-3e311d07c1e944f99f2ba38a21476c46 |
| institution | Directory Open Access Journal |
| issn | 2311-5637 |
| language | English |
| last_indexed | 2024-03-09T03:26:15Z |
| publishDate | 2022-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Fermentation |
| spelling | doaj.art-3e311d07c1e944f99f2ba38a21476c462023-12-03T15:01:39ZengMDPI AGFermentation2311-56372022-06-018729910.3390/fermentation8070299Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i>Xiaomei Zhang0Zhenhang Sun1Jinyu Bian2Yujie Gao3Dong Zhang4Guoqiang Xu5Xiaojuan Zhang6Hui Li7Jinsong Shi8Zhenghong Xu9Laboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaLaboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaLaboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaNational Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, ChinaLaboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaNational Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, ChinaNational Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, ChinaLaboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaLaboratory of Pharmaceutical Engineering, School of Life Science and Health Engineering, Jiangnan University, Wuxi 214122, ChinaNational Engineering Research Center for Cereal Fermentation and Food Biomanufacturing, Jiangnan University, Wuxi 214122, China<span style="font-variant: small-caps;">l</span>-Cysteine is an important sulfur-containing amino acid with numerous applications in the pharmaceutical and cosmetic industries. The microbial production of <span style="font-variant: small-caps;">l</span>-cysteine has received substantial attention, and the supply of the precursor <span style="font-variant: small-caps;">l</span>-serine is important in <span style="font-variant: small-caps;">l</span>-cysteine biosynthesis. In this study, to achieve <span style="font-variant: small-caps;">l</span>-cysteine overproduction, we first increased <span style="font-variant: small-caps;">l</span>-serine production by deleting genes involved in the pathway of <span style="font-variant: small-caps;">l</span>-serine degradation to glycine (serine hydroxymethyl transferase, SHMT, encoded by <i>glyA</i> genes) in strain 4W (with <span style="font-variant: small-caps;">l</span>-serine titer of 1.1 g/L), thus resulting in strain 4WG with <span style="font-variant: small-caps;">l</span>-serine titer of 2.01 g/L. Second, the serine-biosensor based on the transcriptional regulator NCgl0581 of <i>C. glutamicum</i> was constructed in <i>E. coli,</i> and the validity and sensitivity of the biosensor were demonstrated in <i>E. coli</i>. Then 4WG was further evolved through adaptive laboratory evolution (ALE) combined with serine-biosensor, thus yielding the strain 4WGX with 4.13 g/L <span style="font-variant: small-caps;">l</span>-serine production. Moreover, the whole genome of the evolved strain 4WGX was sequenced, and ten non-synonymous mutations were found in the genome of strain 4WGX compared with strain 4W. Finally, 4WGX was used as the starting strain, and deletion of the <span style="font-variant: small-caps;">l</span>-cysteine desulfhydrases (encoded by <i>tnaA</i>), overexpression of serine acetyltransferase (encoded by <i>cysE</i>) and the key enzyme of transport pathway (encoded by <i>ydeD</i>) were performed in strain 4WGX. The recombinant strain 4WGX-∆<i>tnaA</i>-<i>cysE</i>-<i>ydeD</i> can produce 313.4 mg/L of <span style="font-variant: small-caps;">l</span>-cysteine using glycerol as the carbon source. This work provides an efficient method for the biosynthesis of value-added commodity products associated with glycerol conversion.https://www.mdpi.com/2311-5637/8/7/299<i>Escherichia coli</i>biosensorglyceroladaptive laboratory evolution<span style="font-variant: small-caps">l</span>-cysteine |
| spellingShingle | Xiaomei Zhang Zhenhang Sun Jinyu Bian Yujie Gao Dong Zhang Guoqiang Xu Xiaojuan Zhang Hui Li Jinsong Shi Zhenghong Xu Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> Fermentation <i>Escherichia coli</i> biosensor glycerol adaptive laboratory evolution <span style="font-variant: small-caps">l</span>-cysteine |
| title | Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> |
| title_full | Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> |
| title_fullStr | Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> |
| title_full_unstemmed | Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> |
| title_short | Rational Metabolic Engineering Combined with Biosensor-Mediated Adaptive Laboratory Evolution for <span style="font-variant: small-caps">l</span>-Cysteine Overproduction from Glycerol in <i>Escherichia coli</i> |
| title_sort | rational metabolic engineering combined with biosensor mediated adaptive laboratory evolution for span style font variant small caps l span cysteine overproduction from glycerol in i escherichia coli i |
| topic | <i>Escherichia coli</i> biosensor glycerol adaptive laboratory evolution <span style="font-variant: small-caps">l</span>-cysteine |
| url | https://www.mdpi.com/2311-5637/8/7/299 |
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