Multienzymatic cascade synthesis of fucosyloligosaccharide via a two-step fermentation strategy in Escherichia coli

Qin, Hao; Li, Songtao; Zhang, Yufu; Wang, Jianwen; Li, Jixuan; Song, Shiyi; Lu, Fuping; Yu, Li

doi:10.1007/s10529-016-2151-y

Cited by 6 publications

(2 citation statements)

References 15 publications

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“…Enzymatic synthesis is more amenable to larger scale carbohydrate production, but also has its challenges. GTs have successfully been used in the synthesis of HMO structures in vitro (Chen et al, 2015; Yu et al, 2017a), but their application is hampered by the use of expensive nucleotide-activated sugars, and multi-enzyme substrate recycling systems are needed to prevent metabolites from inhibiting enzyme activity (Qin et al, 2016). Using bacterial cells as production factories however, major advancements in HMO production have been made and have resulted in FDA approval and commercialization of the major HMO 2′-fucosyllactose.…”

Section: Development Of Novel Ndcsmentioning

confidence: 99%

Shaping the Infant Microbiome With Non-digestible Carbohydrates

et al. 2019

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Natural polysaccharides with health benefits are characterized by a large structural diversity and differ in building blocks, linkages, and lengths. They contribute to human health by functioning as anti-adhesives preventing pathogen adhesion, stimulate immune maturation and gut barrier function, and serve as fermentable substrates for gut bacteria. Examples of such beneficial carbohydrates include the human milk oligosaccharides (HMOs). Also, specific non-digestible carbohydrates (NDCs), such as galacto-oligosaccharides (GOS) and fructo-oligosaccharides (FOS) are being produced with this purpose in mind, and are currently added to infant formula to stimulate the healthy development of the newborn. They mimic some functions of HMO, but not all. Therefore, many research efforts focus on identification and production of novel types of NDCs. In this review, we give an overview of the few NDCs currently available [GOS, FOS, polydextrose (PDX)], and outline the potential of alternative oligosaccharides, such as pectins, (arabino)xylo-oligosaccharides, and microbial exopolysaccharides (EPS). Moreover, state-of-the-art techniques to generate novel types of dietary glycans, including sialylated GOS (Sia-GOS) and galactosylated chitin, are presented as a way to obtain novel prebiotic NDCs that help shaping the infant microbiome.

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Section: Development Of Novel Ndcsmentioning

confidence: 99%

Shaping the Infant Microbiome With Non-digestible Carbohydrates

et al. 2019

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“…to generate GTP and NADPH (Lee et al 2011, 2012). However, there are drawbacks to microbial fermentation, such as bacterial growth status, complicated separations, and the presence of by-products (Qin et al 2016). As an alternative, enzymatic synthesis provides greater efficiency by specifying GDP- l -fucose enzymes that can be immobilized and reused, thus lowering production costs and limiting the amounts of by-products (Liu et al 2015).…”

Section: Introductionmentioning

confidence: 99%

Cell-free enzymatic synthesis of GDP-l-fucose from mannose

et al. 2019

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GDP- l -fucose, the key substrate for fucosyloligosaccharide biosynthesis, has been synthesized via a de novo pathway in bacteria. In the present study, genes for GDP- l -fucose biosynthesis were cloned into the expression vector pET-28a (+) to construct five E. coli strains, with recombinant enzymes being purified by using Ni–NTA chromatography. Following optimization of the 3-step reaction, Glk, ManB and ManC were added to the reaction mixture, after which Gmd and WcaG were added to overcome feedback inhibition from the end-product to produce GDP- l -fucose at 178.6 mg/l, with a yield of 14.1%. Our studies provide the basis for using cell-free enzyme production of GDP- l -fucose.

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