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Chemoenzymatic synthesis of sialyl Lewis x containing different sialic acid forms
Author(s) -
Huang Ronald Wei,
Lau Kam,
Sugiarto Go,
Thon Vireak,
Li Yanhong,
Ding Li
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.480.3
Subject(s) - tetrasaccharide , chemistry , sialyl lewis x , sialyltransferase , glycan , glycosyltransferase , fucosyltransferase , enzyme , stereochemistry , biochemistry , monosaccharide , sialic acid , lewis acids and bases , azide , glycobiology , glycosylation , selectin , catalysis , organic chemistry , polysaccharide , glycoprotein , adhesion
Found in many animals and microorganisms, sialyl Lewis X tetrasaccharide plays essential roles in inflammatory responses and cancer metastasis. In order to better understand the physiological and pathological importance of sialyl Lewis X, it is essential to obtain sialyl Lewis X with substantial purity. A general, convenient, and efficient enzymatic approach for producing sialyl Lewis x tetrasaccharide is attempted. Initially, a one‐pot three‐enzyme approach, using enzymes alpha‐D‐glucose‐1‐phosphate uridylyltransferase (GalU), UDP‐galactose‐4‐epimerase (GalE), and a beta‐1,4‐glycosyltransferase (LgtB), was employed to form disaccharide LacNAc‐beta‐propyl azide from the monosaccharide substrate GlcNAc‐beta‐propyl azide in the galactosylation reaction. Using a one‐pot two‐enzyme system containing Neisseria meningitidis CMP‐sialic acid synthetase (NmCSS) and Pasteurella multocida sialyltransferase (PmST1), we produced Sialyl‐alpha‐2,3‐LacNAc‐beta‐propyl azide. This sialylation reaction was useful in producing different sialic acid forms. In the final step, sialyl Lewis X was produced using a one‐pot two‐ enzyme fucosylation reaction utilizing alpha‐1,3‐fucosyltransferase (alpha1,3FucT) and L‐fucokinase pyrophosphorylase (FKP). The products will be used to generate glycan arrays for glycomics studies.