Effective Separation of Human Milk Glycosides using Carbon Dioxide Supercritical Fluid Chromatography

Liou, Shih-Wei; Fang, Jia-Lin; Lin, Hung-Wei; Tsai, Teng-Wei; Huang, Hsiao-Yun; Liang, Chin-Yu; Yang, Cheng-Ruel; Wei, Guor‐Tzo; Yu, Ching‐Ching

doi:10.1002/asia.202001404

Cited by 8 publications

(2 citation statements)

References 53 publications

(58 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Treatment of 4b with Bf13FT catalysis resulted in two monofucosylated isomers, LNFP III (59%) and LNnFP V (11%), and unreacted LNnT (29%), as displayed in Table (entry 2). The incubation of 4c with Bf13FT produced two monofucosylated isomers, LNFP III (63%) and LNnFP V (11%), and one difucosylated product, LNDFH III (12%), with 14% of LNnT unreacted (Table , entry 3), as determined by supercritical fluid chromatography . These data indicated that the inherent substrate promiscuity of Bf13FT, which led to the synthesis of several fucosylated isomers from LNnT, could be altered by switching the O -azidohexyl linker to the SF 17 tag.…”

Section: Resultsmentioning

confidence: 93%

“…The incubation of 4c with Bf13FT produced two monofucosylated isomers, LNFP III (63%) and LNnFP V (11%), and one difucosylated product, LNDFH III (12%), with 14% of LNnT unreacted (Table 1, entry 3), as determined by supercritical fluid chromatography. 73 These data indicated that the inherent substrate promiscuity of Bf13FT, which led to the synthesis of several fucosylated isomers from LNnT, could be altered by switching the Oazidohexyl linker to the SF 17 tag. This process increased its substrate specificity ( modification of the anomeric position with a short N-methyl oxyamine linker exhibited no effect on catalytic activity and selectivity enhancement.…”

Section: ■ Results and Discussionmentioning

confidence: 95%

See 1 more Smart Citation

Sulfo-Fluorous Tagging Strategy for Site-Selective Enzymatic Glycosylation of para-Human Milk Oligosaccharides

Huang

et al. 2021

ACS Catal.

Self Cite

View full text Add to dashboard Cite

In this study, we developed an efficient fluorous tagging strategy for site-selective enzymatic fucosylation/sialylation in which appending a readily removable auxiliary tag (SF 17 ) to the reducing end of an acceptor provided the regioselectivity control of a given wild-type fucosyltransferase/sialyltransferase. By combining this strategy with the sequential one-pot enzymatic system of HMO production, various fucosylated para-HMOs, such as LNFP I, LNFP II, LNFP III, LNFP V, LNDFH I, LNDFH II, TF-LNT, F-pLNH I, F-pLNH IV, DF-pLNH I isomer, DF-pLNH II, TF-pLNH I, TF-pLNH II, F-pLNnH, F-pLNnH I, DF-pLNnH, TF-pLNnH, and TetraF-pLNO, were synthesized. Furthermore, the SF 17 -tagged acceptors improved the reaction rates of the enzymatic glycan chain extension. Terminal α2,6-sialylation on SF 17 -tagged pLNnH was directly achieved using wild-type α2,6-sialyltransferase from Photobacterium damsela (Pd26ST). Moreover, enzyme kinetics revealed that the catalytic efficiency (k cat/K M) of α1,4-fucosylation of Bf13FT (α1,3/4-fucosyltransferase from Bacteroides fragilis) on SF 17 -tagged LNFP V acceptors could be enhanced by 55 times compared with tagging with an azidohexyl aglycone. We developed an efficient and reliable strategy for generating complex and diverse natural HMO libraries for functional gut microbiome studies. This SF 17 -assisted glycosylation strategy provides a practical approach to harness the extreme substrate flexibility offered by readily available bacterial glycosyltransferases, which enables the site-selective fucosylation/sialylation on para-HMOs.

show abstract

Section: Resultsmentioning

confidence: 93%

Section: ■ Results and Discussionmentioning

confidence: 95%

Sulfo-Fluorous Tagging Strategy for Site-Selective Enzymatic Glycosylation of para-Human Milk Oligosaccharides

Huang

et al. 2021

ACS Catal.

Self Cite

View full text Add to dashboard Cite

show abstract

Chemical Synthesis of Human Milk Oligosaccharides: para‐Lacto‐N‐hexaose and para‐Lacto‐N‐neohexaose

Singh,

Escopy,

Shadrick

et al. 2023

Chemistry A European J

View full text Add to dashboard Cite

Human milk oligosaccharides (HMO) have emerged as a very active area of research in glycoscience and nutrition. HMO are involved in the early development of infants and may help to prevent certain diseases. The development of chemical methods for obtaining individual HMO aids the global effort dedicated to understanding the roles of these biomolecules. Reported herein is the chemical synthesis of two common core hexasaccharides found in human milk, i. e. para‐lacto‐N‐hexaose (pLNH) and para‐lacto‐N‐neohexaose (pLNnH). After screening multiple leaving groups and temporary protecting group combinations, a 3+3 convergent coupling strategy was found to work best for obtaining these linear glycans.

show abstract

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

Harvey

2024

Mass Spectrometry Reviews

View full text Add to dashboard Cite

The use of matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry for the analysis of carbohydrates and glycoconjugates is a well‐established technique and this review is the 12th update of the original article published in 1999 and brings coverage of the literature to the end of 2022. As with previous review, this review also includes a few papers that describe methods appropriate to analysis by MALDI, such as sample preparation, even though the ionization method is not MALDI. The review follows the same format as previous reviews. It is divided into three sections: (1) general aspects such as theory of the MALDI process, matrices, derivatization, MALDI imaging, fragmentation, quantification and the use of computer software for structural identification. (2) Applications to various structural types such as oligo‐ and polysaccharides, glycoproteins, glycolipids, glycosides and biopharmaceuticals, and (3) other general areas such as medicine, industrial processes, natural products and glycan synthesis where MALDI is extensively used. Much of the material relating to applications is presented in tabular form. MALDI is still an ideal technique for carbohydrate analysis, particularly in its ability to produce single ions from each analyte and advancements in the technique and range of applications show little sign of diminishing.

show abstract

Effective Separation of Human Milk Glycosides using Carbon Dioxide Supercritical Fluid Chromatography

Cited by 8 publications

References 53 publications

Sulfo-Fluorous Tagging Strategy for Site-Selective Enzymatic Glycosylation of para-Human Milk Oligosaccharides

Sulfo-Fluorous Tagging Strategy for Site-Selective Enzymatic Glycosylation of para-Human Milk Oligosaccharides

Chemical Synthesis of Human Milk Oligosaccharides: para‐Lacto‐N‐hexaose and para‐Lacto‐N‐neohexaose

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update for 2021–2022

Contact Info

Product

Resources

About