Synthesis of the milk oligosaccharide 2′-fucosyllactose using recombinant bacterial enzymes

Albermann, Christoph; Piepersberg, Wolfgang; Wehmeier, Udo F.

doi:10.1016/s0008-6215(01)00177-x

Cited by 86 publications

(73 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…[8] Thus, in this study the enzyme was generated in stand-alone form and exhibited a specific activity comparable to the reported protein fusion. [17] Although the enzyme has been demonstrated to be soluble, [13] the expression level and specific activity found in this study were much lower compared to the values published for the a1,3- [14] and a1,4-counterparts. [19] However, the H. pylori a1,2-FucT appeared to be a more effective fucosyltransferase, as displayed by the kinetic results (Table 2), when compared to the recently investigated E. coli a1,2-FucT (WbsJ).…”

Section: Discussioncontrasting

confidence: 74%

“…[17] In contrast, other studies have not been able to reproduce this effect. [8] Thus, in this study the enzyme was generated in stand-alone form and exhibited a specific activity comparable to the reported protein fusion.…”

Section: Discussionmentioning

confidence: 90%

“…[13] Slight variations of this bias could be observed for certain H. pylori source strains. [16] H. pylori a1,2-FucT has been employed for the synthesis of 2'-fucosyllactose [17] and H-type 2 [8] products in vitro and in vivo, respectively, without gaining further information about its selectivity and potency.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of Helicobacter pylori α1,2‐Fucosyltransferase for Enzymatic Synthesis of Tumor‐Associated Antigens

Stein

Lin

2008

Adv Synth Catal

View full text Add to dashboard Cite

The a1,2-fucosyltransferase (a1,2-FucT) from Helicobacter pylori catalyzes the fucosylation of acceptor oligosaccharides at the C2-OH of terminal Galb units. The enzyme from strain NCTC11639 was evaluated for its ability to synthesize cancer-associated antigens. The a1,2-FucT was determined to be active over a pH range between 4.0 and 8.0 with the optimum occurring at pH 5.0. Although a divalent metal ion cofactor was not required for catalysis, enhancement of the enzyme activity was detected upon supplement with Mn 2+ . Detailed substrate specificity analysis revealed that a1,2-FucT can catalyze the fucosylation of a wide variety of oligosaccharide substrates. The a1,2-FucT preferentially fucosylated type 1 structure (Galb1-3GlcNAc)-containing glycans over type 2 structure (Galb1-4GlcNAc)-containing glycans. The Lewis a trisaccharide [Galb1-3-

show abstract

Section: Discussioncontrasting

confidence: 74%

Section: Discussionmentioning

confidence: 90%

See 1 more Smart Citation

Characterization of Helicobacter pylori α1,2‐Fucosyltransferase for Enzymatic Synthesis of Tumor‐Associated Antigens

Stein

Lin

2008

Adv Synth Catal

View full text Add to dashboard Cite

show abstract

“…There are two types of enzymes that can carry out fucosylation: fucosyltransferases and fucosidases. Fucosyltransferases present high specificity toward the acceptor and do not hydrolyze the product (11,12). However, those enzymes are generally difficult to express and they require an expensive sugar nucleotide or a complex multienzymatic system for the regeneration (11,13).…”

mentioning

confidence: 99%

“…Fucosyltransferases present high specificity toward the acceptor and do not hydrolyze the product (11,12). However, those enzymes are generally difficult to express and they require an expensive sugar nucleotide or a complex multienzymatic system for the regeneration (11,13). In contrast, oligosaccharides have been largely produced using the transglycosylation activity of glycosidases and engineered glycosynthases (14,15).…”

mentioning

confidence: 99%

Synthesis of Fucosyl- N -Acetylglucosamine Disaccharides by Transfucosylation Using α-l-Fucosidases from Lactobacillus casei

Rodrı́guez-Dı́az

Carbajo²,

Pineda‐Lucena³

et al. 2013

Appl Environ Microbiol

View full text Add to dashboard Cite

bAlfB and AlfC ␣-L-fucosidases from Lactobacillus casei were used in transglycosylation reactions, and they showed high efficiency in synthesizing fucosyldisaccharides. AlfB and AlfC activities exclusively produced fucosyl-␣-1,3-N-acetylglucosamine and fucosyl-␣-1,6-N-acetylglucosamine, respectively. The reaction kinetics showed that AlfB can convert 23% p-nitrophenyl-␣-L-fucopyranoside into fucosyl-␣-1,3-N-acetylglucosamine and AlfC at up to 56% into fucosyl-␣-1,6-N-acetylglucosamine. Fucosyl-oligosaccharides (FUS) are coming to be of great interest due to their presence in human milk and their antiadhesive activity against pathogens. This last property is characterized by blocking the attachment of intestinal pathogens to host cells due to FUS structural similarity with cell-surface glycoconjugate receptors (1-4). In addition, genes encoding ␣-L-fucosidases (EC 3.2.1.51) have recently been found in genomes of bifidobacteria (5, 6) and lactobacilli (7), which raises issues regarding the role of these enzymes in the metabolism of fucose-containing structures and opens up the possibility of using FUS as novel prebiotics (8).To fully establish the biological function of FUS as antiadhesins and prebiotics, extensive studies are required, for which synthesis of sufficient amounts would be necessary. Chemical synthesis of FUS such as ABH and Lewis blood group antigens has long been performed, but it is a tedious and expensive process since it requires multiple protection and deprotection steps to achieve the desired selectivity (9, 10). The enzymatic synthesis of FUS is an alternative to chemical methods, and it offers the advantage of forming specific glycosidic linkages in the presence of other reactive functional groups. There are two types of enzymes that can carry out fucosylation: fucosyltransferases and fucosidases. Fucosyltransferases present high specificity toward the acceptor and do not hydrolyze the product (11, 12). However, those enzymes are generally difficult to express and they require an expensive sugar nucleotide or a complex multienzymatic system for the regeneration (11, 13). In contrast, oligosaccharides have been largely produced using the transglycosylation activity of glycosidases and engineered glycosynthases (14,15). Only a few works describe the synthesis of FUS by transglycosylation using ␣-L-fucosidases; however, they cover a wide range of sources such as mammalian tissues (16), fungi (17), and bacteria (18,19). The transglycosylation activity of ␣-L-fucosidases is generally moderate compared with the hydrolysis activity, although it is variable and depends on the origin of the enzymes (20).We have previously isolated three ␣-L-fucosidases (AlfA, AlfB, and AlfC) from Lactobacillus casei that cleaved ␣-linked L-fucose from natural FUS (7). AlfB and AlfC showed high specific activity on fucosyl-␣-1,3-N-acetylglucosamine (Fuc-␣-1,3-GlcNAc) and fucosyl-␣-1,6-N-acetylglucosamine (Fuc-␣-1,6-GlcNAc), respectively. These substrate specificities led us in the present work to explore the likely us...

show abstract

Food Oligosaccharides

Moreno¹,

Sanz²

2014

View full text Add to dashboard Cite

Synthesis of the milk oligosaccharide 2′-fucosyllactose using recombinant bacterial enzymes

Cited by 86 publications

References 24 publications

Characterization of Helicobacter pylori α1,2‐Fucosyltransferase for Enzymatic Synthesis of Tumor‐Associated Antigens

Characterization of Helicobacter pylori α1,2‐Fucosyltransferase for Enzymatic Synthesis of Tumor‐Associated Antigens

Synthesis of Fucosyl- N -Acetylglucosamine Disaccharides by Transfucosylation Using α-l-Fucosidases from Lactobacillus casei

Food Oligosaccharides

Contact Info

Product

Resources

About