Journal: Applied Microbiology and Biotechnology

Article Title: Synthesis of fucosyllactose using α-L-fucosidases GH29 from infant gut microbial metagenome

doi: 10.1007/s00253-024-13178-3

Figure Lengend Snippet: HPLC chromatograms (Jasco PU2080Plus coupled to a refractive index detector) of Fuc2358 ( a ) and Fuc5372 ( b ) transfucosylation reactions, 1, Tris-HCl; 2, F1 or F2 fraction containing fucosyllactose (FL) with the same retention time as the 2′-fucosyllactose (2′FL) standard; 3, lactose; 4, mixture of L-fucose and glycerol. HPAEC-PAD chromatograms (Dionex ICS3000 system) of the purified fractions F1 from Fuc2358 and F2 from Fuc5372 ( c ), and 2′FL, 6′fucosyllactose (6’FL), 2′,3-difucosyllactose (DFL), and 3-fucosyllactose (3FL) standards. The numbers indicate the retention times of the peaks in minutes

Article Snippet: The synthesized oligosaccharides contained in fractions F1 and F2 were confirmed by comparison of their retention times with those of standards 2′FL, 3FL, DFL, and 6′-fucosyllactose (6′FL) (Biosynth AG, Staad, Switzerland) by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) in a Dionex ICS3000 system using a CarboPacTMPA100 analytical column equipped with a CarboPacTMPA100 guard column (Dionex Corp., Sunnyvale, CA, USA).

Techniques: Refractive Index, Purification

Journal: Applied Microbiology and Biotechnology

Article Title: Synthesis of fucosyllactose using α-L-fucosidases GH29 from infant gut microbial metagenome

doi: 10.1007/s00253-024-13178-3

Figure Lengend Snippet: 1 H 1D NMR spectra of commercial 2′-fucosyllactose (2′FL) ( a ) and fractions F1 ( b ) and F2 ( c ) from Fuc2358 and Fuc5372, respectively. Spectra were acquired at 600 MHz. ns = 256. T = 25 °C in D 2 O. Signals corresponding to 2′FL compound are marked in blue. Common signals between F1 and F2 are marked in red and could be assigned to 3′-fucosyllactose (3′FL)

Article Snippet: The synthesized oligosaccharides contained in fractions F1 and F2 were confirmed by comparison of their retention times with those of standards 2′FL, 3FL, DFL, and 6′-fucosyllactose (6′FL) (Biosynth AG, Staad, Switzerland) by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) in a Dionex ICS3000 system using a CarboPacTMPA100 analytical column equipped with a CarboPacTMPA100 guard column (Dionex Corp., Sunnyvale, CA, USA).

Techniques:

Journal: Applied Microbiology and Biotechnology

Article Title: Synthesis of fucosyllactose using α-L-fucosidases GH29 from infant gut microbial metagenome

doi: 10.1007/s00253-024-13178-3

Figure Lengend Snippet: a LC-ESI-MS chromatograms of transfucosylation reactions with Fuc2358 and Fuc5372, and 2′FL standard. b Fuc2358 purified fractions f1 and f2. c Fuc2358 purified fraction f3. d Fuc5372 purified fractions f1 and f2. e Fuc5372 purified fraction f3. f MS 2 fragmentation patterns of m/z 487.0 Da of the 2′FL standard and f2 and f3 peaks of Fuc2358 and Fuc5372. The parent ion [M-H]− is indicated by a diamond. For each fraction, the untreated fraction (top), the fraction treated with NaBH 4 (middle), and the fraction incubated with the β-galactosidase from Aspergillus oryzae (bottom) are represented. Isomers present in the samples without treatment or not affected by the treatment are represented in black, reduced isomers are represented in green, and isomers hydrolyzed by the β-galactosidase are represented in red. The relative percentage of the fucosyllactose isomers produced is represented ( a ). All LC-ESI-MS chromatograms are combined extraction ion chromatograms (EIC) of deprotonated [M-H] − and formate adducts [M-H + FA] − . 2′FL, 2′-fucosyllactose; 3′FL, 3′-fucosyllactose; 1FL, 1-fucosyllactose; 2/6FL, 2/6-fucosyllactose

Article Snippet: The synthesized oligosaccharides contained in fractions F1 and F2 were confirmed by comparison of their retention times with those of standards 2′FL, 3FL, DFL, and 6′-fucosyllactose (6′FL) (Biosynth AG, Staad, Switzerland) by high-pH anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD) in a Dionex ICS3000 system using a CarboPacTMPA100 analytical column equipped with a CarboPacTMPA100 guard column (Dionex Corp., Sunnyvale, CA, USA).

Techniques: Purification, Incubation, Produced, Extraction