Synergetic engineering of central carbon and nitrogen metabolism for the production of N-acetylglucosamine in Bacillus subtilis.
Bacillus subtilis
N-acetylglucosamine
glutamate
glutamine
gudB
rocG
Journal
Biotechnology and applied biochemistry
ISSN: 1470-8744
Titre abrégé: Biotechnol Appl Biochem
Pays: United States
ID NLM: 8609465
Informations de publication
Date de publication:
Jan 2020
Jan 2020
Historique:
received:
14
08
2019
accepted:
23
10
2019
pubmed:
28
10
2019
medline:
15
12
2020
entrez:
27
10
2019
Statut:
ppublish
Résumé
N-acetylglucosamine (GlcNAc) is a nitrogen-containing compound, which is widely used as a nutraceutical and pharmaceutical. In our previous work, we constructed a recombinant Bacillus subtilis strain for the biosynthesis of GlcNAc by engineering the central carbon metabolism. However, nitrogen is also required for the synthesis of GlcNAc. Hence, it is necessary to simultaneously coordinate the carbon and nitrogen metabolism to improve production of GlcNAc. In this work, we attempted to enhance GlcNAc production in B. subtilis by increasing supply of precursors N-acetylglucosamine 6-phosphate (GlcNAc6P) and glutamate. The expression of a key enzyme, GlcNAc6P N-acetyltransferase (GNA1), was enhanced by engineering the promoter and ribosome binding site to enhance the production of GlcNAc6P. Next, we examined the effect of different nitrogen sources on GlcNAc synthesis. We observed that urea can promote nitrogen assimilation for GlcNAc synthesis. The glutamate synthesis was improved by deleting the two endogenous glutamate dehydrogenase genes (rocG and gudB) and by integrating one exogenous glutamate dehydrogenase gene (gdh). This strategy enhanced the intracellular glutamate and glutamine by 69.8% and 46.9%, respectively. The synergetic engineering of central carbon and nitrogen metabolisms increased the GlcNAc titer from 14.0 to 22.2 g/L in the shaker flask. Hence, our study demonstrated the importance of carbon and nitrogen metabolism coordination in the production of nitrogen-containing compounds.
Substances chimiques
Carbon
7440-44-0
Nitrogen
N762921K75
Acetylglucosamine
V956696549
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
123-132Subventions
Organisme : National Natural Science Foundation of China
ID : 31622001
Organisme : National Natural Science Foundation of China
ID : 31671845
Organisme : National Natural Science Foundation of China
ID : 21676119
Organisme : National Natural Science Foundation of China
ID : 31600068
Organisme : Natural Science Foundation of Jiangsu Province
ID : BK20160176
Organisme : Overseas Expertise Introduction Project for Discipline Innovation
ID : 111-2-06
Organisme : National Natural Science Foundation of China
ID : 21676119,
Informations de copyright
© 2019 International Union of Biochemistry and Molecular Biology, Inc.
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