Engineering a colanic acid biosynthesis pathway in E. coli for manufacturing 2’-fucosyllactose

Qiao, Chunming; Wu, Haiying; Ji, Mengyao; Xie, Yukang; Li, Sijie; Li, Yiqing; Shi, Jiping; Sun, Junsong

doi:10.1016/j.procbio.2020.04.017

Cited by 12 publications

(14 citation statements)

References 34 publications

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“…Eleven new α1,2-fucosyltransferase candidates were tested for in vivo synthesis of 2′-FL through the de novo GDP- l -fucose pathway, and only the enzyme from B. fragilis NCTC9343, WcfB, succeeded . In another study, WcfB was also determined to be effective, but less effective than FutC, for 2′-FL synthesis in engineered E. coli . In addition, Te2FT was also screened from four α1,2-fucosyltransferase candidates, which could efficiently produce 2′-FL in engineered E. coli …”

Section: α12-fucosyltransferasementioning

confidence: 99%

“…Several enzymes are involved in the colanic acid pathway from GDP- l -fucose, in which UDP-glucose:undecaprenyl-phosphate glucose-1-phosphate transferase (WcaJ, EC 2.7.8.31) is the enzyme responsible for the first step . Deletion of competitive pathway gene wcaJ was widely performed to prevent GDP- l -fucose loss and further improve 2′-FL production in engineered E. coli . ,,,− ,, In addition, Lee et al studied the effect of additional deletion of waaF encoding for ADP-heptose:LPS heptosyltransferase II, which might influence the GDP- l -fucose transport. Deletion of waaF was confirmed to favor colanic acid synthesis .…”

Section: ′-Fl Biosynthesis By Metabolically Engineered Strainsmentioning

confidence: 99%

“…Codon optimization, which is a regular and simple approach to improve the expression of heterologous proteins, has widely been used for α1,2-fucosyltransferase expression for 2′-FL synthesis in E. coli . ,, However, it only increases the transcriptional level but does not improve the solubility during the translation process. Promoter selection was tried to regulate the functional expression of α1,2-fucosyltransferase for optimization of 2′-FL synthesis . Decreased promoter strength might enhance the α1,2-fucosyltransferase solubility in E. coli , leading to improved 2′-FL synthesis …”

Section: ′-Fl Biosynthesis By Metabolically Engineered Strainsmentioning

confidence: 99%

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Strategies for Enhancing Microbial Production of 2′-Fucosyllactose, the Most Abundant Human Milk Oligosaccharide

Liu

Zhu

Wang³

et al. 2022

J. Agric. Food Chem.

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Human milk oligosaccharides (HMOs), a group of structurally diverse unconjugated glycans in breast milk, act as important prebiotics and have plenty of unique health effects for growing infants. 2′-Fucosyllactose (2′-FL) is the most abundant HMO, accounting for approximately 30%, among approximately 200 identified HMOs with different structures. 2′-FL can be enzymatically produced by α1,2-fucosyltransferase, using GDP-l-fucose as donor and lactose as acceptor. Metabolic engineering strategies have been widely used for enhancement of GDP-l-fucose supply and microbial production of 2′-FL with high productivity. GDP-l-fucose supply can be enhanced by two main pathways, including de novo and salvage pathways. 2′-FL-producing α1,2-fucosyltransferases have widely been identified from various microorganisms. Metabolic pathways for 2′-FL synthesis can be basically constructed by enhancing GDP-l-fucose supply and introducing α1,2-fucosyltransferase. Various strategies have been attempted to enhance 2′-FL production, such as acceptor enhancement, donor enhancement, and improvement of the functional expression of α1,2-fucosyltransferase. In this review, current progress in GDP-l-fucose synthesis and bacterial α1,2-fucosyltransferases is described in detail, various metabolic engineering strategies for enhancing 2′-FL production are comprehensively reviewed, and future research focuses in biotechnological production of 2′-FL are suggested.

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Section: α12-fucosyltransferasementioning

confidence: 99%

Section: ′-Fl Biosynthesis By Metabolically Engineered Strainsmentioning

confidence: 99%

Section: ′-Fl Biosynthesis By Metabolically Engineered Strainsmentioning

confidence: 99%

See 1 more Smart Citation

Strategies for Enhancing Microbial Production of 2′-Fucosyllactose, the Most Abundant Human Milk Oligosaccharide

Liu

Zhu

Wang³

et al. 2022

J. Agric. Food Chem.

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“…This approach increased the production to 47.0 g/L by eliminating arabinose isomerase (araA) and rhamnose isomerase (rhaA), and using fucose, lactose, and glycerol as substitutes. Recently, Chen et al 71 reported that colonic acid (CA) synthesis using E. coli S17−3 as host strain was triggered by low pH. This pathway is catalyzed in vivo by heterologous-1,2-fucosyltransferases to generate GDP-L-fucose, producing 0.62 g/L of 2′-FL.…”

Section: Hmos Production Using Microbial Systemsmentioning

confidence: 99%

Engineered Microbial Routes for Human Milk Oligosaccharides Synthesis

Mosleh

Abbaspourrad

2021

ACS Synth. Biol.

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Human milk oligosaccharides (HMOs) are one of the important ingredients in human milk, which have attracted great interest due to their beneficial effect on the health of newborns. The large-scale production of HMOs has been researched using engineered microbial routes due to the availability, safety, and low cost of host strains. In addition, the development of molecular biology technology and metabolic engineering has promoted the effectiveness of HMOs production. According to current reports, 2′-fucosyllactose (2′-FL), 3-fucosyllactose (3-FL), lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), 3′-sialyllactose (3′-SL), 6′-sialyllactose (6′-SL), and some fucosylated HMOs with complex structures have been produced via the engineered microbial route, with 2′-FL having been produced the most. However, due to the uncertainty of metabolic patterns, the selection of host strains has certain limitations. Aside from that, the expression of appropriate glycosyltransferase in microbes is key to the synthesis of different HMOs. Therefore, finding a safe and efficient glycosyltransferase has to be addressed when using engineered microbial pathways. In this review, the latest research on the production of HMOs using engineered microbial routes is reported. The selection of host strains and adapting different metabolic pathways helped researchers designing engineered microbial routes that are more conducive to HMOs production.

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“…Increasing the expression of guanosine inosine kinase (Gsk) is another approach to boosting GDP- l -fucose accumulation, as Gsk can optimize the supply of guanosine 5′-triphosphate (GTP), which is the rate-limiting substrate of Fkp . Along the salvage pathway, it has been well documented that reducing or blocking lactose and l -fucose metabolism in the E. coli chassis possessing α-1,2-fucosyltransferase and Fkp can markedly increase the 2′-FL yield. , After the α-1,2-fucosyltransferase gene derived from H. pylori was introduced to recombine the E. coli BL21star (DE3) strain assimilating lactose, the 2′-FL yield by the recombinant strain remained low. When the same gene was expressed in E. coli JM109 (DE3), the highest 2′-FL titer of 1.23 g/L was obtained with 14.5 g/L lactose .…”

Section: Microbial Production Of 2′-flmentioning

confidence: 99%

Biotechnological Production of 2′-Fucosyllactose: A Prevalent Fucosylated Human Milk Oligosaccharide

et al. 2021

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Human milk oligosaccharide (HMO) is a key component of human milk carbohydrates and is closely related to the nutrition and health benefits of breastfeeding in infants. 2′-Fucosyllactose (2′-FL) is the most abundant fucosylated HMO, which has remarkable value in nutrition and medicine, such as suppressing pathogen infection, regulating intestinal flora, and boosting immunity. However, 2′-FL production via the method of extraction or chemical synthesis cannot meet its large demand, and as a result, environmentally friendly and efficient biotechnological approaches, including in vitro enzymatic synthesis and microbial cell factory production, have been developed and applied to its commercialized production. This review introduces, summarizes, and discusses the recent advances in the biotechnological production of 2′-FL. Furthermore, future research directions for the biotechnological production of 2′-FL as well as the strategies to further improve its concentration are highlighted and discussed.

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Engineering a colanic acid biosynthesis pathway in E. coli for manufacturing 2’-fucosyllactose

Cited by 12 publications

References 34 publications

Strategies for Enhancing Microbial Production of 2′-Fucosyllactose, the Most Abundant Human Milk Oligosaccharide

Strategies for Enhancing Microbial Production of 2′-Fucosyllactose, the Most Abundant Human Milk Oligosaccharide

Engineered Microbial Routes for Human Milk Oligosaccharides Synthesis

Biotechnological Production of 2′-Fucosyllactose: A Prevalent Fucosylated Human Milk Oligosaccharide

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