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Microbial production of human milk oligosaccharide lactodifucotetraose
- Source :
- Metabolic Engineering. 66:12-20
- Publication Year :
- 2021
- Publisher :
- Elsevier BV, 2021.
-
Abstract
- Human milk oligosaccharides (HMOs) are potent bioactive compounds that modulate neonatal health and are of interest for development as potential drug treatments for adult diseases. The potential of these molecules, their limited access from natural sources, and difficulty in large-scale isolation of individual HMOs for studies and applications have motivated the development of chemical syntheses and in vitro enzymatic catalysis strategies. Whole cell biocatalysts are emerging as alternative self-regulating production platforms that have the potential to reduce the cost for enzymatic synthesis of HMOs. Whole cell biocatalysts for the production of short-chained, linear and small monofucosylated HMOs have been reported but those for fucosylated structures with higher complexity have not been explored. In this study, we established a strategy for producing a difucosylated HMO, lactodifucotetraose (LDFT), from lactose and L-fucose in Escherichia coli. We used two bacterial fucosyltransferases with narrow acceptor selectivity to drive the sequential fucosylation of lactose and intermediate 2′-fucosyllactose (2′-FL) to produce LDFT. Deletion of substrate degradation pathways that decoupled cellular growth from LDFT production, enhanced expression of native substrate transporters and modular induction of the genes in the LDFT biosynthetic pathway allowed complete conversion of lactose into LDFT and minor quantities of the side product 3-fucosyllactose (3-FL). Overall, 5.1 g/L of LDFT was produced from 3 g/L lactose and 3 g/L L-fucose in 24 h. Our results demonstrate promising applications of engineered microbial biosystems for the production of multi-fucosylated HMOs for biochemical studies.
- Subjects :
- 0106 biological sciences
Oligosaccharides
Bioengineering
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
Enzyme catalysis
Metabolic engineering
Fucosyltransferases
03 medical and health sciences
chemistry.chemical_compound
010608 biotechnology
medicine
Humans
Lactose
Escherichia coli
Fucosylation
Fucose
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
Milk, Human
Oligosaccharide
In vitro
chemistry
Biochemistry
Biotechnology
Subjects
Details
- ISSN :
- 10967176
- Volume :
- 66
- Database :
- OpenAIRE
- Journal :
- Metabolic Engineering
- Accession number :
- edsair.doi.dedup.....aeb55e4dd770c83832fe6c37451cc70d