Biotechnological production of fucosylated human milk oligosaccharides: Prokaryotic fucosyltransferases and their use in biocatalytic cascades or whole cell conversion systems

Petschacher, Barbara; Nidetzky, Bernd

doi:10.1016/j.jbiotec.2016.03.052

Cited by 97 publications

(86 citation statements)

References 98 publications

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“…While metabolic engineering of Escherichia coli has recently been shown to allow direct microbial production of appreciable fucosyllactose (FL) yields [8][9][10], production of structurally more complex HMOs in engineered E. coli is still in its infancy [10,11]. As a simpler alternative, direct enzymatic regiospecific synthesis of fucosylated HMOs in vitro employing α-L-fucosidases to catalyze transfucosylation is an option [12][13][14][15].…”

Section: Accepted M M a N U mentioning

confidence: 99%

Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides

et al. 2018

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Human milk oligosaccharides (HMOs) constitute a unique family of bioactive lactose-based molecules present in human breast milk. HMOs are of major importance for infant health and development but also virtually absent from bovine milk used for infant formula. Among the HMOs, the fucosylated species are the most abundant. Transfucosylation catalysed by retaining α-l-fucosidases is a new route for manufacturing biomimetic HMOs. Seven α-l-fucosidases from glycosyl hydrolase family 29 were expressed, characterized in terms of substrate specificity and thermal stability, and shown to be able to catalyse transfucosylation. The α-l-1,3/4-fucosidase CpAfc2 from Clostridium perfringens efficiently catalysed the formation of the more complex human milk oligosaccharide structure lacto-N-fucopentaose II (LNFP II) using 3-fucosyllactose as fucosyl donor and lacto-N-tetraose as acceptor with a 39% yield. α-l-Fucosidases FgFCO1 from Fusarium graminearum and Mfuc5 from a soil metagenome were able to catalyse transfucosylation of lactose using citrus xyloglucan as fucosyl donor. FgFCO1 catalysed formation of 2'-fucosyllactose, whereas Mfuc5 catalysis mainly produced an unidentified, non-HMO fucosyllactose, reaching molar yields based on the donor substrate of 14% and 18%, respectively.

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Section: Accepted M M a N U mentioning

confidence: 99%

Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides

et al. 2018

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“…In biosynthesis of 2‐FL using engineered E. coli , lactose is fucosylated by exogenous fucosyltransferase using guanosine 5′‐diphosphate (GDP)‐ l ‐fucose as a fucose donor (Lee et al, ). GDP‐ l ‐fucose is synthesized in the cytoplasm through the de novo pathway or the salvage pathway (Petschacher & Nidetzky, ). The de novo pathway is catalyzed by GDP‐ d ‐mannose 4,6‐dehydratase (Gmd) and GDP‐ l ‐fucose synthase (WcaG) using GDP‐ d ‐mannose from central metabolism (Bulter & Elling, ).…”

mentioning

confidence: 99%

Enhanced production of 2’‐fucosyllactose from fucose by elimination of rhamnose isomerase and arabinose isomerase in engineered Escherichia coli

Jung

Chin

Lee

et al. 2019

Biotech & Bioengineering

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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.

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“…However, their application in preparative synthesis, which aims to produce the compound of interest on a large scale, is restricted due to the low availability of glycosyltransferases and their donor substrates. In addition, the latter are difficult to synthesize and are therefore costly .…”

Section: Employment Of Fucosidases In the Synthesis Of Fucosylated Olmentioning

confidence: 99%

“…This bacterium produces, in low quantities, GDP‐Fuc, a fucosyl‐nucleotide that is suitable for use as a donor substrate with fucosyltransferases. Some strategies attempt to overexpress, delete, or insert genes that encode key enzymes in the de novo and salvage pathways of synthesis of GDP‐Fuc to increase its concentration in the cell . In a similar manner, some efforts have been conducted to transform E. coli into a “cell factory” for fucooligosaccharides, being capable of synthesizing not only the donor substrate in reasonable amounts, but also the acceptor substrate and a suitable fucosyltransferase for the synthesis of a specific compound.…”

Section: Employment Of Fucosidases In the Synthesis Of Fucosylated Olmentioning

confidence: 99%

Employment of fucosidases for the synthesis of fucosylated oligosaccharides with biological potential

Guzmán-Rodríguez

Alatorre‐Santamaría

Gómez-Ruiz

et al. 2018

Biotech and App Biochem

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Fucosylated oligosaccharides play important physiological roles in humans, including in the immune response, transduction of signals, early embryogenesis and development, growth regulation, apoptosis, pathogen adhesion, and so on. Efforts have been made to synthesize fucosylated oligosaccharides, as it is difficult to purify them from their natural sources, such as human milk, epithelial tissue, blood, and so on. Within the strategies for its in vitro synthesis, it is remarkable the employment of fucosidases, enzymes that normally cleave the fucosyl residue from the non-reducing end of fucosylated compounds, as these enzymes are also capable of synthesizing them by means of a transfucosylation reaction. This review summarizes the progress in the use of fucosidases for the synthesis of compounds that have potential for industrial and commercial applications.

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Biotechnological production of fucosylated human milk oligosaccharides: Prokaryotic fucosyltransferases and their use in biocatalytic cascades or whole cell conversion systems

Cited by 97 publications

References 98 publications

Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides

Substrate specificity and transfucosylation activity of GH29 α-l-fucosidases for enzymatic production of human milk oligosaccharides

Enhanced production of 2’‐fucosyllactose from fucose by elimination of rhamnose isomerase and arabinose isomerase in engineered Escherichia coli

Employment of fucosidases for the synthesis of fucosylated oligosaccharides with biological potential

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