A practical approach to N-glycan production by hydrazinolysis using hydrazine monohydrate.
Hydrazinolysis is a versatile method to liberate N-linked glycans from glycoproteins. However, the method is usually performed with anhydrous hydrazine, a highly toxic and explosive chemical used in rocket fuel. Thus despite the need to produce functionally important glyco-materials, hydrazinolysis is limited to small scale (e.g., 0.2-1 mL) reactions. In the present study, we report an alternative procedure for hydrazinolysis using hydrazine monohydrate in place of anhydrous hydrazine. The developed procedure was applied to both purified glycoproteins (Taka-amylase and transferrin) and hen egg yolk protein fraction with comparable yields to the traditional method using anhydrous hydrazine. The sialyl linkage of alpha2-6disialobiantennary oligosaccharides proved to be fully stable. The developed procedure facilitated the large-scale preparation of N-linked glycans. The new method should make a substantial contribution to both small- and large-scale production of functional glycans, including therapeutically relevant human-type glycans.
Nakakita
Shin-ichi
S
Department of Functional Glycomics, Life Science Research Center, Kagawa University, 1750-1 Ikenobe, Kita-gun, Kagawa 761-0793, Japan.
Sumiyoshi
Wataru
W
Miyanishi
Nobumitsu
N
Hirabayashi
Jun
J
D000818:Animals; D001671:Biochemistry / Q000295:instrumentation / Q000379:methods*; D002240:Carbohydrate Sequence; D002645:Chickens; D004530:Egg Yolk / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry; D006026:Glycoside Hydrolases / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D014168:Transferrin / Q000737:chemistry; D000516:alpha-Amylases / Q000737:chemistry
D006023:Glycoproteins; D006834:Hydrazines; D009844:Oligosaccharides; D011134:Polysaccharides; D014168:Transferrin; C029424:hydrazine; D006026:Glycoside Hydrolases; D000516:alpha-Amylases