Synthetic redesign of central carbon and redox metabolism for high yield production of N-acetylglucosamine in Bacillus subtilis
File(s)SE180119JZY_manuscript_final.docx (9.11 MB)
Accepted version
Author(s)
Type
Journal Article
Abstract
One of the primary goals of microbial metabolic engineering is to achieve high titer, yield and productivity (TYP) of engineered strains. This TYP index requires optimized carbon flux toward desired molecule with minimal by-product formation. De novo redesign of central carbon and redox metabolism holds great promise to alleviate pathway bottleneck and improve carbon and energy utilization efficiency. The engineered strain, with the overexpression or deletion of multiple genes, typically can’t meet the TYP index, due to overflow of central carbon and redox metabolism that compromise the final yield, despite a high titer or productivity might be achieved. To solve this challenge, we reprogramed the central carbon and redox metabolism of Bacillus subtilis and achieved high TYP production of N-acetylglucosamine. Specifically, a “push–pull–promote” approach efficiently reduced the overflown acetyl-CoA flux and eliminated byproduct formation. Four synthetic NAD(P)-independent metabolic routes were introduced to rewire the redox metabolism to minimize energy loss. Implementation of these genetic strategies led us to obtain a B. subtilis strain with superior TYP index. GlcNAc titer in shake flask was increased from 6.6 g L−1 to 24.5 g L−1, the yield was improved from 0.115 to 0.468 g GlcNAc g−1 glucose, and the productivity was increased from 0.274 to 0.437 g L−1 h−1. These titer and yield are the highest levels ever reported and, the yield reached 98% of the theoretical pathway yield (0.478 g g−1 glucose). The synthetic redesign of carbon metabolism and redox metabolism represent a novel and general metabolic engineering strategy to improve the performance of microbial cell factories.
Date Issued
2019-01-01
Date Acceptance
2018-10-01
Citation
Metabolic Engineering, 2019, 51, pp.59-69
ISSN
1096-7176
Publisher
Elsevier
Start Page
59
End Page
69
Journal / Book Title
Metabolic Engineering
Volume
51
Copyright Statement
© 2019 Elsevier Ltd. All rights reserved. This manuscript is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licence http://creativecommons.org/licenses/by-nc-nd/4.0/
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000456420200007&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Subjects
Science & Technology
Life Sciences & Biomedicine
Biotechnology & Applied Microbiology
Metabolic engineering
N-acetylglucosamine
Redox metabolism
Overflow metabolism
Bacillus subtilis
Microbial cell factories
Synthetic biology
DYNAMIC PATHWAY REGULATION
MICROBIAL-PRODUCTION
ESCHERICHIA-COLI
TCA CYCLE
CHEMICALS
SYSTEM
FLUX
GLUCOSAMINE
STRATEGIES
REGULATOR
Publication Status
Published
Date Publish Online
2018-10-19