PMID- 25000293
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20211021
IS  - 2160-4762 (Electronic)
IS  - 2160-4762 (Linking)
VI  - 4
DP  - 2012 Sep 1
TI  - One-pot multienzyme synthesis of Lewis x and sialyl Lewis x antigens.
PG  - 233-247
AB  - L-Fucose has been found abundantly in human milk oligosaccharides, bacterial 
      lipopolysaccharides, glycolipids, and many N- and O-linked glycans produced by 
      mammalian cells. Fucose-containing carbohydrates have important biological 
      functions. Alterations in the expression of fucosylated oligosaccharides have 
      been observed in several pathological processes such as cancer and 
      atherosclerosis. Chemical formation of fucosidic bonds is challenging due to its 
      acid lability. Enzymatic construction of fucosidic bonds by fucosyltransferases 
      is highly efficient and selective but requires the expensive sugar nucleotide 
      donor guanosine 5'- diphosphate-L-fucose (GDP-Fuc). Here, we describe a protocol 
      for applying a one-pot three-enzyme system in synthesizing structurally defined 
      fucose-containing oligosaccharides from free L-fucose. In this system, GDP-Fuc is 
      generated from L-fucose, adenosine 5'-triphosphate (ATP), and guanosine 
      5'-triphosphate (GTP) by a bifunctional L-fucokinase/GDP-fucose pyrophosphorylase 
      (FKP). An inorganic pyrophosphatase (PpA) is used to degrade the by-product 
      pyrophosphate (PPi) to drive the reaction towards the formation of GDP-Fuc. In 
      situ generated GDP-Fuc is then used by a suitable fucosyltransferase for the 
      formation of fucosides. The three-enzyme reactions are carried out in one pot 
      without the need for high cost sugar nucleotide or isolation of intermediates. 
      The time for the synthesis is 4-24 hours. Purification and characterization of 
      products can be completed in 2-3 days.
FAU - Yu, Hai
AU  - Yu H
AD  - Department of Chemistry, University of California-Davis, One Shields Avenue, 
      Davis, CA 95616 USA.
FAU - Lau, Kam
AU  - Lau K
AD  - Department of Chemistry, University of California-Davis, One Shields Avenue, 
      Davis, CA 95616 USA.
FAU - Li, Yanhong
AU  - Li Y
AD  - Department of Chemistry, University of California-Davis, One Shields Avenue, 
      Davis, CA 95616 USA.
FAU - Sugiarto, Go
AU  - Sugiarto G
AD  - Department of Chemistry, University of California-Davis, One Shields Avenue, 
      Davis, CA 95616 USA.
FAU - Chen, Xi
AU  - Chen X
AD  - Department of Chemistry, University of California-Davis, One Shields Avenue, 
      Davis, CA 95616 USA.
LA  - eng
GR  - R01 HD065122/HD/NICHD NIH HHS/United States
PT  - Journal Article
PL  - United States
TA  - Curr Protoc Chem Biol
JT  - Current protocols in chemical biology
JID - 101559235
PMC - PMC4076701
MID - NIHMS405289
OTO - NOTNLM
OT  - FKP
OT  - Lewis x
OT  - enzymatic synthesis
OT  - fucose
OT  - fucoside
OT  - fucosylation
OT  - fucosyltransferase
OT  - one-pot multienzyme
OT  - sialyl Lewis x
EDAT- 2012/09/01 00:00
MHDA- 2012/09/01 00:01
PMCR- 2014/07/01
CRDT- 2014/07/08 06:00
PHST- 2014/07/08 06:00 [entrez]
PHST- 2012/09/01 00:00 [pubmed]
PHST- 2012/09/01 00:01 [medline]
PHST- 2014/07/01 00:00 [pmc-release]
AID - 10.1002/9780470559277.ch110277 [doi]
PST - ppublish
SO  - Curr Protoc Chem Biol. 2012 Sep 1;4:233-247. doi: 10.1002/9780470559277.ch110277.