Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.
Identifieur interne : 000135 ( Main/Exploration ); précédent : 000134; suivant : 000136Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.
Auteurs : Jiangming Zhu [République populaire de Chine] ; Wei Yang [République populaire de Chine] ; Bohua Wang [République populaire de Chine] ; Qun Liu [République populaire de Chine] ; Xiaotong Zhong [République populaire de Chine] ; Quanxiu Gao [République populaire de Chine] ; Jiezheng Liu [République populaire de Chine] ; Jianzhong Huang [République populaire de Chine] ; Baixue Lin [République populaire de Chine] ; Yong Tao [République populaire de Chine]Source :
- Microbial cell factories [ 1475-2859 ] ; 2020.
Abstract
BACKGROUND
L-Alanyl-L-glutamine (AQ) is a functional dipeptide with high water solubility, good thermal stability and high bioavailability. It is widely used in clinical treatment, post-operative rehabilitation, sports health care and other fields. AQ is mainly produced via chemical synthesis which is complicated, time-consuming, labor-intensive, and have a low yield accompanied with the generation of by-products. It is therefore highly desirable to develop an efficient biotechnological process for the industrial production of AQ.
RESULTS
A metabolically engineered E. coli strain for AQ production was developed by over-expressing L-amino acid α-ligase (BacD) from Bacillus subtilis, and inactivating the peptidases PepA, PepB, PepD, and PepN, as well as the dipeptide transport system Dpp. In order to use the more readily available substrate glutamic acid, a module for glutamine synthesis from glutamic acid was constructed by introducing glutamine synthetase (GlnA). Additionally, we knocked out glsA-glsB to block the first step in glutamine metabolism, and glnE-glnB involved in the ATP-dependent addition of AMP/UMP to a subunit of glutamine synthetase, which resulted in increased glutamine supply. Then the glutamine synthesis module was combined with the AQ synthesis module to develop the engineered strain that uses glutamic acid and alanine for AQ production. The expression of BacD and GlnA was further balanced to improve AQ production. Using the final engineered strain p15/AQ10 as a whole-cell biocatalyst, 71.7 mM AQ was produced with a productivity of 3.98 mM/h and conversion rate of 71.7%.
CONCLUSION
A metabolically engineered strain for AQ production was successfully developed via inactivation of peptidases, screening of BacD, introduction of glutamine synthesis module, and balancing the glutamine and AQ synthesis modules to improve the yield of AQ. This work provides a microbial cell factory for efficient production of AQ with industrial potential.
DOI: 10.1186/s12934-020-01369-2
PubMed: 32527330
PubMed Central: PMC7291740
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.</title><author><name sortKey="Zhu, Jiangming" sort="Zhu, Jiangming" uniqKey="Zhu J" first="Jiangming" last="Zhu">Jiangming Zhu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Wei" sort="Yang, Wei" uniqKey="Yang W" first="Wei" last="Yang">Wei Yang</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Bohua" sort="Wang, Bohua" uniqKey="Wang B" first="Bohua" last="Wang">Bohua Wang</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Qun" sort="Liu, Qun" uniqKey="Liu Q" first="Qun" last="Liu">Qun Liu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Zhong, Xiaotong" sort="Zhong, Xiaotong" uniqKey="Zhong X" first="Xiaotong" last="Zhong">Xiaotong Zhong</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Gao, Quanxiu" sort="Gao, Quanxiu" uniqKey="Gao Q" first="Quanxiu" last="Gao">Quanxiu Gao</name><affiliation wicri:level="1"><nlm:affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian</wicri:regionArea><wicri:noRegion>Fujian</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Jiezheng" sort="Liu, Jiezheng" uniqKey="Liu J" first="Jiezheng" last="Liu">Jiezheng Liu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Huang, Jianzhong" sort="Huang, Jianzhong" uniqKey="Huang J" first="Jianzhong" last="Huang">Jianzhong Huang</name><affiliation wicri:level="1"><nlm:affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China. hjz@fjnu.edu.cn.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian</wicri:regionArea><wicri:noRegion>Fujian</wicri:noRegion></affiliation></author><author><name sortKey="Lin, Baixue" sort="Lin, Baixue" uniqKey="Lin B" first="Baixue" last="Lin">Baixue Lin</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. linbaixue@126.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China. linbaixue@126.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Tao, Yong" sort="Tao, Yong" uniqKey="Tao Y" first="Yong" last="Tao">Yong Tao</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2020">2020</date><idno type="RBID">pubmed:32527330</idno><idno type="pmid">32527330</idno><idno type="doi">10.1186/s12934-020-01369-2</idno><idno type="pmc">PMC7291740</idno><idno type="wicri:Area/Main/Corpus">000104</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Corpus" wicri:corpus="PubMed">000104</idno><idno type="wicri:Area/Main/Curation">000104</idno><idno type="wicri:explorRef" wicri:stream="Main" wicri:step="Curation">000104</idno><idno type="wicri:Area/Main/Exploration">000104</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.</title><author><name sortKey="Zhu, Jiangming" sort="Zhu, Jiangming" uniqKey="Zhu J" first="Jiangming" last="Zhu">Jiangming Zhu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Yang, Wei" sort="Yang, Wei" uniqKey="Yang W" first="Wei" last="Yang">Wei Yang</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Wang, Bohua" sort="Wang, Bohua" uniqKey="Wang B" first="Bohua" last="Wang">Bohua Wang</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Qun" sort="Liu, Qun" uniqKey="Liu Q" first="Qun" last="Liu">Qun Liu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Zhong, Xiaotong" sort="Zhong, Xiaotong" uniqKey="Zhong X" first="Xiaotong" last="Zhong">Xiaotong Zhong</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Gao, Quanxiu" sort="Gao, Quanxiu" uniqKey="Gao Q" first="Quanxiu" last="Gao">Quanxiu Gao</name><affiliation wicri:level="1"><nlm:affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian</wicri:regionArea><wicri:noRegion>Fujian</wicri:noRegion></affiliation></author><author><name sortKey="Liu, Jiezheng" sort="Liu, Jiezheng" uniqKey="Liu J" first="Jiezheng" last="Liu">Jiezheng Liu</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Huang, Jianzhong" sort="Huang, Jianzhong" uniqKey="Huang J" first="Jianzhong" last="Huang">Jianzhong Huang</name><affiliation wicri:level="1"><nlm:affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China. hjz@fjnu.edu.cn.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian</wicri:regionArea><wicri:noRegion>Fujian</wicri:noRegion></affiliation></author><author><name sortKey="Lin, Baixue" sort="Lin, Baixue" uniqKey="Lin B" first="Baixue" last="Lin">Baixue Lin</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. linbaixue@126.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China. linbaixue@126.com.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author><author><name sortKey="Tao, Yong" sort="Tao, Yong" uniqKey="Tao Y" first="Yong" last="Tao">Yong Tao</name><affiliation wicri:level="1"><nlm:affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101</wicri:regionArea><wicri:noRegion>100101</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</nlm:affiliation><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>University of Chinese Academy of Sciences, Beijing, 100049</wicri:regionArea><wicri:noRegion>100049</wicri:noRegion></affiliation></author></analytic><series><title level="j">Microbial cell factories</title><idno type="eISSN">1475-2859</idno><imprint><date when="2020" type="published">2020</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p><b>BACKGROUND</b></p><p>L-Alanyl-L-glutamine (AQ) is a functional dipeptide with high water solubility, good thermal stability and high bioavailability. It is widely used in clinical treatment, post-operative rehabilitation, sports health care and other fields. AQ is mainly produced via chemical synthesis which is complicated, time-consuming, labor-intensive, and have a low yield accompanied with the generation of by-products. It is therefore highly desirable to develop an efficient biotechnological process for the industrial production of AQ.</p></div><div type="abstract" xml:lang="en"><p><b>RESULTS</b></p><p>A metabolically engineered E. coli strain for AQ production was developed by over-expressing L-amino acid α-ligase (BacD) from Bacillus subtilis, and inactivating the peptidases PepA, PepB, PepD, and PepN, as well as the dipeptide transport system Dpp. In order to use the more readily available substrate glutamic acid, a module for glutamine synthesis from glutamic acid was constructed by introducing glutamine synthetase (GlnA). Additionally, we knocked out glsA-glsB to block the first step in glutamine metabolism, and glnE-glnB involved in the ATP-dependent addition of AMP/UMP to a subunit of glutamine synthetase, which resulted in increased glutamine supply. Then the glutamine synthesis module was combined with the AQ synthesis module to develop the engineered strain that uses glutamic acid and alanine for AQ production. The expression of BacD and GlnA was further balanced to improve AQ production. Using the final engineered strain p15/AQ10 as a whole-cell biocatalyst, 71.7 mM AQ was produced with a productivity of 3.98 mM/h and conversion rate of 71.7%.</p></div><div type="abstract" xml:lang="en"><p><b>CONCLUSION</b></p><p>A metabolically engineered strain for AQ production was successfully developed via inactivation of peptidases, screening of BacD, introduction of glutamine synthesis module, and balancing the glutamine and AQ synthesis modules to improve the yield of AQ. This work provides a microbial cell factory for efficient production of AQ with industrial potential.</p></div></front></TEI><pubmed><MedlineCitation Status="In-Process" Owner="NLM"><PMID Version="1">32527330</PMID><DateRevised><Year>2020</Year><Month>06</Month><Day>15</Day></DateRevised><Article PubModel="Electronic"><Journal><ISSN IssnType="Electronic">1475-2859</ISSN><JournalIssue CitedMedium="Internet"><Volume>19</Volume><Issue>1</Issue><PubDate><Year>2020</Year><Month>Jun</Month><Day>11</Day></PubDate></JournalIssue><Title>Microbial cell factories</Title><ISOAbbreviation>Microb Cell Fact</ISOAbbreviation></Journal><ArticleTitle>Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.</ArticleTitle><Pagination><MedlinePgn>129</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1186/s12934-020-01369-2</ELocationID><Abstract><AbstractText Label="BACKGROUND" NlmCategory="BACKGROUND">L-Alanyl-L-glutamine (AQ) is a functional dipeptide with high water solubility, good thermal stability and high bioavailability. It is widely used in clinical treatment, post-operative rehabilitation, sports health care and other fields. AQ is mainly produced via chemical synthesis which is complicated, time-consuming, labor-intensive, and have a low yield accompanied with the generation of by-products. It is therefore highly desirable to develop an efficient biotechnological process for the industrial production of AQ.</AbstractText><AbstractText Label="RESULTS" NlmCategory="RESULTS">A metabolically engineered E. coli strain for AQ production was developed by over-expressing L-amino acid α-ligase (BacD) from Bacillus subtilis, and inactivating the peptidases PepA, PepB, PepD, and PepN, as well as the dipeptide transport system Dpp. In order to use the more readily available substrate glutamic acid, a module for glutamine synthesis from glutamic acid was constructed by introducing glutamine synthetase (GlnA). Additionally, we knocked out glsA-glsB to block the first step in glutamine metabolism, and glnE-glnB involved in the ATP-dependent addition of AMP/UMP to a subunit of glutamine synthetase, which resulted in increased glutamine supply. Then the glutamine synthesis module was combined with the AQ synthesis module to develop the engineered strain that uses glutamic acid and alanine for AQ production. The expression of BacD and GlnA was further balanced to improve AQ production. Using the final engineered strain p15/AQ10 as a whole-cell biocatalyst, 71.7 mM AQ was produced with a productivity of 3.98 mM/h and conversion rate of 71.7%.</AbstractText><AbstractText Label="CONCLUSION" NlmCategory="CONCLUSIONS">A metabolically engineered strain for AQ production was successfully developed via inactivation of peptidases, screening of BacD, introduction of glutamine synthesis module, and balancing the glutamine and AQ synthesis modules to improve the yield of AQ. This work provides a microbial cell factory for efficient production of AQ with industrial potential.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Zhu</LastName><ForeName>Jiangming</ForeName><Initials>J</Initials><Identifier Source="ORCID">http://orcid.org/0000-0001-8996-2086</Identifier><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Yang</LastName><ForeName>Wei</ForeName><Initials>W</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Wang</LastName><ForeName>Bohua</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Qun</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Zhong</LastName><ForeName>Xiaotong</ForeName><Initials>X</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Gao</LastName><ForeName>Quanxiu</ForeName><Initials>Q</Initials><AffiliationInfo><Affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Liu</LastName><ForeName>Jiezheng</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Huang</LastName><ForeName>Jianzhong</ForeName><Initials>J</Initials><AffiliationInfo><Affiliation>National Engineering Research Center of Industrial Microbiology and Fermentation Technology, College of Life Sciences, Fujian Normal University, Fuzhou, 350117, Fujian, People's Republic of China. hjz@fjnu.edu.cn.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Lin</LastName><ForeName>Baixue</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China. linbaixue@126.com.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China. linbaixue@126.com.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Tao</LastName><ForeName>Yong</ForeName><Initials>Y</Initials><AffiliationInfo><Affiliation>Chinese Academy of Sciences Key Laboratory of Microbial Physiological and Metabolic Engineering, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>University of Chinese Academy of Sciences, Beijing, 100049, China.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><GrantList CompleteYN="Y"><Grant><GrantID>31670051</GrantID><Agency>National Natural Science Foundation of China</Agency><Country></Country></Grant><Grant><GrantID>2018YFA0901400</GrantID><Agency>the National Key Research and Development Program of China</Agency><Country></Country></Grant></GrantList><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2020</Year><Month>06</Month><Day>11</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>Microb Cell Fact</MedlineTA><NlmUniqueID>101139812</NlmUniqueID><ISSNLinking>1475-2859</ISSNLinking></MedlineJournalInfo><CitationSubset>IM</CitationSubset><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="N">AQ</Keyword><Keyword MajorTopicYN="N">Glutamine synthase</Keyword><Keyword MajorTopicYN="N">L-amino acid α-ligase</Keyword><Keyword MajorTopicYN="N">Metabolic engineering</Keyword><Keyword MajorTopicYN="N">Whole-cell biocatalysis</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2019</Year><Month>11</Month><Day>28</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2020</Year><Month>05</Month><Day>16</Day></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2020</Year><Month>6</Month><Day>13</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>epublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">32527330</ArticleId><ArticleId IdType="doi">10.1186/s12934-020-01369-2</ArticleId><ArticleId IdType="pii">10.1186/s12934-020-01369-2</ArticleId><ArticleId IdType="pmc">PMC7291740</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Appl Biochem Biotechnol. 2017 Sep;183(1):362-373</Citation><ArticleIdList><ArticleId IdType="pubmed">28337691</ArticleId></ArticleIdList></Reference><Reference><Citation>Bioresour Technol. 2017 Dec;245(Pt B):1603-1609</Citation><ArticleIdList><ArticleId IdType="pubmed">28624247</ArticleId></ArticleIdList></Reference><Reference><Citation>Am J Physiol Gastrointest Liver Physiol. 2015 May 15;308(10):G831-9</Citation><ArticleIdList><ArticleId IdType="pubmed">25792564</ArticleId></ArticleIdList></Reference><Reference><Citation>Nutr Clin Pract. 2016 Aug;31(4):445-50</Citation><ArticleIdList><ArticleId IdType="pubmed">27246308</ArticleId></ArticleIdList></Reference><Reference><Citation>Microbiology. 1999 Oct;145 ( Pt 10):2891-901</Citation><ArticleIdList><ArticleId IdType="pubmed">10537211</ArticleId></ArticleIdList></Reference><Reference><Citation>Int J Pept Protein Res. 1990 Mar;35(3):161-214</Citation><ArticleIdList><ArticleId IdType="pubmed">2191922</ArticleId></ArticleIdList></Reference><Reference><Citation>Amino Acids. 1992 Oct;3(3):293-302</Citation><ArticleIdList><ArticleId IdType="pubmed">24193129</ArticleId></ArticleIdList></Reference><Reference><Citation>Nutrition. 1997 Jul-Aug;13(7-8):731-7</Citation><ArticleIdList><ArticleId IdType="pubmed">9263278</ArticleId></ArticleIdList></Reference><Reference><Citation>Clin Nutr. 2004 Feb;23(1):13-21</Citation><ArticleIdList><ArticleId IdType="pubmed">14757388</ArticleId></ArticleIdList></Reference><Reference><Citation>Nucleic Acids Res. 2017 May 19;45(9):5437-5448</Citation><ArticleIdList><ArticleId IdType="pubmed">28158713</ArticleId></ArticleIdList></Reference><Reference><Citation>EMBO J. 1997 Sep 15;16(18):5562-71</Citation><ArticleIdList><ArticleId IdType="pubmed">9312015</ArticleId></ArticleIdList></Reference><Reference><Citation>Nucleic Acids Res. 2014 Feb;42(4):2646-59</Citation><ArticleIdList><ArticleId IdType="pubmed">24234441</ArticleId></ArticleIdList></Reference><Reference><Citation>Pol Tyg Lek. 1980 Aug 18;35(33):1275-8</Citation><ArticleIdList><ArticleId IdType="pubmed">7433211</ArticleId></ArticleIdList></Reference><Reference><Citation>J Am Chem Soc. 2016 Jun 8;138(22):7016-23</Citation><ArticleIdList><ArticleId IdType="pubmed">27199273</ArticleId></ArticleIdList></Reference><Reference><Citation>Biomed Res Int. 2015;2015:545467</Citation><ArticleIdList><ArticleId IdType="pubmed">26495301</ArticleId></ArticleIdList></Reference><Reference><Citation>Biotechnol Appl Biochem. 2011 Nov-Dec;58(6):449-55</Citation><ArticleIdList><ArticleId IdType="pubmed">22172107</ArticleId></ArticleIdList></Reference><Reference><Citation>Mol Microbiol. 1994 Dec;14(5):1077-92</Citation><ArticleIdList><ArticleId IdType="pubmed">7536291</ArticleId></ArticleIdList></Reference><Reference><Citation>Proc Natl Acad Sci U S A. 1979 Sep;76(9):4544-8</Citation><ArticleIdList><ArticleId IdType="pubmed">41243</ArticleId></ArticleIdList></Reference><Reference><Citation>J Bacteriol. 2005 Aug;187(15):5195-202</Citation><ArticleIdList><ArticleId IdType="pubmed">16030213</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochemistry. 2008 May 27;47(21):5724-35</Citation><ArticleIdList><ArticleId IdType="pubmed">18459799</ArticleId></ArticleIdList></Reference><Reference><Citation>J Biotechnol. 2005 Jan 26;115(2):211-20</Citation><ArticleIdList><ArticleId IdType="pubmed">15607239</ArticleId></ArticleIdList></Reference><Reference><Citation>Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):134-42</Citation><ArticleIdList><ArticleId IdType="pubmed">22100191</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Protoc. 2013 Nov;8(11):2281-2308</Citation><ArticleIdList><ArticleId IdType="pubmed">24157548</ArticleId></ArticleIdList></Reference><Reference><Citation>Appl Environ Microbiol. 2007 Oct;73(20):6378-85</Citation><ArticleIdList><ArticleId IdType="pubmed">17720844</ArticleId></ArticleIdList></Reference><Reference><Citation>Amino Acids. 2017 Dec;49(12):2023-2031</Citation><ArticleIdList><ArticleId IdType="pubmed">28861626</ArticleId></ArticleIdList></Reference><Reference><Citation>World J Surg. 2000 Dec;24(12):1503-13</Citation><ArticleIdList><ArticleId IdType="pubmed">11193715</ArticleId></ArticleIdList></Reference><Reference><Citation>Nat Methods. 2009 May;6(5):343-5</Citation><ArticleIdList><ArticleId IdType="pubmed">19363495</ArticleId></ArticleIdList></Reference><Reference><Citation>Microb Cell Fact. 2017 Jun 13;16(1):106</Citation><ArticleIdList><ArticleId IdType="pubmed">28610636</ArticleId></ArticleIdList></Reference><Reference><Citation>Biosci Biotechnol Biochem. 2013;77(3):618-23</Citation><ArticleIdList><ArticleId IdType="pubmed">23470770</ArticleId></ArticleIdList></Reference><Reference><Citation>Protein Expr Purif. 2005 May;41(1):207-34</Citation><ArticleIdList><ArticleId IdType="pubmed">15915565</ArticleId></ArticleIdList></Reference><Reference><Citation>Biochemistry. 2007 Nov 13;46(45):12979-96</Citation><ArticleIdList><ArticleId IdType="pubmed">17939683</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed><affiliations><list><country><li>République populaire de Chine</li></country></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Zhu, Jiangming" sort="Zhu, Jiangming" uniqKey="Zhu J" first="Jiangming" last="Zhu">Jiangming Zhu</name></noRegion><name sortKey="Gao, Quanxiu" sort="Gao, Quanxiu" uniqKey="Gao Q" first="Quanxiu" last="Gao">Quanxiu Gao</name><name sortKey="Huang, Jianzhong" sort="Huang, Jianzhong" uniqKey="Huang J" first="Jianzhong" last="Huang">Jianzhong Huang</name><name sortKey="Lin, Baixue" sort="Lin, Baixue" uniqKey="Lin B" first="Baixue" last="Lin">Baixue Lin</name><name sortKey="Lin, Baixue" sort="Lin, Baixue" uniqKey="Lin B" first="Baixue" last="Lin">Baixue Lin</name><name sortKey="Liu, Jiezheng" sort="Liu, Jiezheng" uniqKey="Liu J" first="Jiezheng" last="Liu">Jiezheng Liu</name><name sortKey="Liu, Jiezheng" sort="Liu, Jiezheng" uniqKey="Liu J" first="Jiezheng" last="Liu">Jiezheng Liu</name><name sortKey="Liu, Qun" sort="Liu, Qun" uniqKey="Liu Q" first="Qun" last="Liu">Qun Liu</name><name sortKey="Liu, Qun" sort="Liu, Qun" uniqKey="Liu Q" first="Qun" last="Liu">Qun Liu</name><name sortKey="Tao, Yong" sort="Tao, Yong" uniqKey="Tao Y" first="Yong" last="Tao">Yong Tao</name><name sortKey="Tao, Yong" sort="Tao, Yong" uniqKey="Tao Y" first="Yong" last="Tao">Yong Tao</name><name sortKey="Wang, Bohua" sort="Wang, Bohua" uniqKey="Wang B" first="Bohua" last="Wang">Bohua Wang</name><name sortKey="Yang, Wei" sort="Yang, Wei" uniqKey="Yang W" first="Wei" last="Yang">Wei Yang</name><name sortKey="Zhong, Xiaotong" sort="Zhong, Xiaotong" uniqKey="Zhong X" first="Xiaotong" last="Zhong">Xiaotong Zhong</name><name sortKey="Zhong, Xiaotong" sort="Zhong, Xiaotong" uniqKey="Zhong X" first="Xiaotong" last="Zhong">Xiaotong Zhong</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Bois/explor/DetoxFungiV1/Data/Main/Exploration
HfdSelect -h $EXPLOR_STEP/biblio.hfd -nk 000135 | SxmlIndent | more
Ou
HfdSelect -h $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd -nk 000135 | SxmlIndent | more
Pour mettre un lien sur cette page dans le réseau Wicri
{{Explor lien
|wiki= Bois
|area= DetoxFungiV1
|flux= Main
|étape= Exploration
|type= RBID
|clé= pubmed:32527330
|texte= Metabolic engineering of Escherichia coli for efficient production of L-alanyl-L-glutamine.
}}
Pour générer des pages wiki
HfdIndexSelect -h $EXPLOR_AREA/Data/Main/Exploration/RBID.i -Sk "pubmed:32527330" \
| HfdSelect -Kh $EXPLOR_AREA/Data/Main/Exploration/biblio.hfd \
| NlmPubMed2Wicri -a DetoxFungiV1
| This area was generated with Dilib version V0.6.38. |  |