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Enhanced production of 2’‐fucosyllactose from fucose by elimination of rhamnose isomerase and arabinose isomerase in engineeredEscherichia coli
- Source :
- Biotechnology and Bioengineering. 116:2412-2417
- Publication Year :
- 2019
- Publisher :
- Wiley, 2019.
-
Abstract
- 2'-Fucosyllactose (2-FL), one of the most abundant oligosaccharides in human milk, has been spotlighted for its neutraceutical and pharmaceutical potentials. Microbial production of 2-FL is promising since it is efficient as compared to other production methods. In 2-FL microbial production via the salvage pathway for biosynthesis of guanosine 5'-diphosphate (GDP)-l-fucose from fucose, the conversion yield from fucose is important because of the high price of fucose. In this study, deletion of the genes (araA and rhaA) coding for arabinose isomerase (AraA) and rhamnose isomerase (RhaA) was attempted in engineered Escherichia coli for improving 2-FL production by using fucose, lactose, and glycerol. The engineered E. coli constructed previously is able to express fucokinase/GDP-l-fucose pyrophosphorylase (Fkp) from Bacteroides fragilis and the α-1,2-fucosyltransferase (FucT2) from Helicobacter pylori and deficient in β-galactosidase (LacZ), fucose isomerase (FucI), and fuculose kinase (FucK). The additional double-deletion of the araA and rhaA genes in the engineered E. coli enhanced the product yield of 2-FL to 0.52 mole 2-FL/mole fucose, and hence the concentration of 2-FL reached to 47.0 g/L, which are 44% and two-fold higher than those (23.1 g/L and 0.36 mole 2-FL/mole fucose) of the control strain in fed-batch fermentation. Elimination of sugar isomerases exhibiting promiscuous activities with fucose might be critical in the microbial production of 2-FL through the salvage pathway of GDP-l-fucose.
- Subjects :
- 0106 biological sciences
0301 basic medicine
Rhamnose
Bioengineering
Isomerase
medicine.disease_cause
01 natural sciences
Applied Microbiology and Biotechnology
Fucose
03 medical and health sciences
chemistry.chemical_compound
2'-Fucosyllactose
010608 biotechnology
Escherichia coli
medicine
Fuculose
Escherichia coli Proteins
030104 developmental biology
Metabolic Engineering
chemistry
Biochemistry
Arabinose isomerase
Fucokinase
Trisaccharides
Gene Deletion
Biotechnology
Subjects
Details
- ISSN :
- 10970290 and 00063592
- Volume :
- 116
- Database :
- OpenAIRE
- Journal :
- Biotechnology and Bioengineering
- Accession number :
- edsair.doi.dedup.....66df71e46bfa0baab1c9cf2144c864c5