Synthetic metabolic channel by functional membrane microdomains for compartmentalized flux control
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
The anchoring of metabolic pathway enzymes to spatial scaffolds can significantly improve their reaction efficiency. Here, we successfully constructed a multi-enzyme complex assembly system able to enhance bioproduction in bacteria by using the endogenous spatial scaffolds─functional membrane microdomains (FMMs). First, using VA-TIRFM and SPT analysis, we reveal that FMMs possess high temporal and spatial stability at the plasma membrane and can be used as endogenous spatial scaffolds to organize enzyme pathways. Then, taking the synthesis of N-acetylglucosamine (GlcNAc) in Bacillus subtilis as a proof-of-concept demonstration, we found that anchoring of various enzymes required for GlcNAc synthesis onto FMMs to obtain the FMMs-multi-enzyme complex system resulted in a significant increase in GlcNAc titer and an effectively alleviate in cell lysis at the later stage of fermentation compared to that in control strains expressing the related enzymes in the cytoplasm. Combining with metabolic model and kinetics analysis, the existence of a constructed substrate channel that maximizes the reaction efficiency is verified. In summary, we propose a novel metabolic pathway assembly model which allowed improved titers and compartmentalized flux control with high spatial resolution in bacterial metabolism.
Date Issued
2020-05-01
Date Acceptance
2020-02-17
Citation
Metabolic Engineering, 2020, 59, pp.106-118
ISSN
1096-7176
Publisher
Elsevier
Start Page
106
End Page
118
Journal / Book Title
Metabolic Engineering
Volume
59
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/32105784
PII: S1096-7176(20)30044-6
Subjects
Bacillus subtilis
Functional membrane microdomains
GlcNAc synthesis
Metabolic channel
Multienzyme complex system
Publication Status
Published
Coverage Spatial
Belgium
Date Publish Online
2020-02-24