One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it>
<p>Abstract</p> <p>Background</p> <p>L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In <it>Escherichia coli</it>, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzym...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-03-01
|
| Series: | Microbial Cell Factories |
| Online Access: | http://www.microbialcellfactories.com/content/11/1/30 |
| _version_ | 1818386276448993280 |
|---|---|
| author | Gu Pengfei Yang Fan Kang Junhua Wang Qian Qi Qingsheng |
| author_facet | Gu Pengfei Yang Fan Kang Junhua Wang Qian Qi Qingsheng |
| author_sort | Gu Pengfei |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In <it>Escherichia coli</it>, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyrophosphate are also involved in L-tryptophan synthesis. In order to construct a microbial strain for efficient L-tryptophan production from glucose, we developed a one step tryptophan attenuator inactivation and promoter swapping strategy for metabolic flux optimization after a base strain was obtained by overexpressing the <it>tktA</it>, mutated <it>trpE </it>and <it>aroG </it>genes and inactivating a series of competitive steps.</p> <p>Results</p> <p>The engineered <it>E. coli </it>GPT1002 with tryptophan attenuator inactivation and tryptophan operon promoter substitution exhibited 1.67 ~ 9.29 times higher transcription of tryptophan operon genes than the control GPT1001. In addition, this strain accumulated 1.70 g l<sup>-1 </sup>L-tryptophan after 36 h batch cultivation in 300-mL shake flask. Bioreactor fermentation experiments showed that GPT1002 could produce 10.15 g l<sup>-1 </sup>L-tryptophan in 48 h.</p> <p>Conclusions</p> <p>The one step inactivating and promoter swapping is an efficient method for metabolic engineering. This method can also be applied in other bacteria.</p> |
| first_indexed | 2024-12-14T03:51:29Z |
| format | Article |
| id | doaj.art-c2cd6f5a358246159cca63d1a3c28f3b |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-14T03:51:29Z |
| publishDate | 2012-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-c2cd6f5a358246159cca63d1a3c28f3b2022-12-21T23:18:12ZengBMCMicrobial Cell Factories1475-28592012-03-011113010.1186/1475-2859-11-30One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it>Gu PengfeiYang FanKang JunhuaWang QianQi Qingsheng<p>Abstract</p> <p>Background</p> <p>L-tryptophan is an aromatic amino acid widely used in the food, chemical and pharmaceutical industries. In <it>Escherichia coli</it>, L-tryptophan is synthesized from phosphoenolpyruvate and erythrose 4-phosphate by enzymes in the shikimate pathway and L-tryptophan branch pathway, while L-serine and phosphoribosylpyrophosphate are also involved in L-tryptophan synthesis. In order to construct a microbial strain for efficient L-tryptophan production from glucose, we developed a one step tryptophan attenuator inactivation and promoter swapping strategy for metabolic flux optimization after a base strain was obtained by overexpressing the <it>tktA</it>, mutated <it>trpE </it>and <it>aroG </it>genes and inactivating a series of competitive steps.</p> <p>Results</p> <p>The engineered <it>E. coli </it>GPT1002 with tryptophan attenuator inactivation and tryptophan operon promoter substitution exhibited 1.67 ~ 9.29 times higher transcription of tryptophan operon genes than the control GPT1001. In addition, this strain accumulated 1.70 g l<sup>-1 </sup>L-tryptophan after 36 h batch cultivation in 300-mL shake flask. Bioreactor fermentation experiments showed that GPT1002 could produce 10.15 g l<sup>-1 </sup>L-tryptophan in 48 h.</p> <p>Conclusions</p> <p>The one step inactivating and promoter swapping is an efficient method for metabolic engineering. This method can also be applied in other bacteria.</p>http://www.microbialcellfactories.com/content/11/1/30 |
| spellingShingle | Gu Pengfei Yang Fan Kang Junhua Wang Qian Qi Qingsheng One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> Microbial Cell Factories |
| title | One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> |
| title_full | One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> |
| title_fullStr | One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> |
| title_full_unstemmed | One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> |
| title_short | One-step of tryptophan attenuator inactivation and promoter swapping to improve the production of L-tryptophan in <it>Escherichia coli</it> |
| title_sort | one step of tryptophan attenuator inactivation and promoter swapping to improve the production of l tryptophan in it escherichia coli it |
| url | http://www.microbialcellfactories.com/content/11/1/30 |
| work_keys_str_mv | AT gupengfei onestepoftryptophanattenuatorinactivationandpromoterswappingtoimprovetheproductionofltryptophaninitescherichiacoliit AT yangfan onestepoftryptophanattenuatorinactivationandpromoterswappingtoimprovetheproductionofltryptophaninitescherichiacoliit AT kangjunhua onestepoftryptophanattenuatorinactivationandpromoterswappingtoimprovetheproductionofltryptophaninitescherichiacoliit AT wangqian onestepoftryptophanattenuatorinactivationandpromoterswappingtoimprovetheproductionofltryptophaninitescherichiacoliit AT qiqingsheng onestepoftryptophanattenuatorinactivationandpromoterswappingtoimprovetheproductionofltryptophaninitescherichiacoliit |