Separation of enantiomers on a chiral stationary phase based on ovoglycoprotein - VIII. Chiral recognition ability of partially and completely deglycosylated ovoglycoprotein

被引:7
作者
Haginaka, J
Matsunaga, H
Kakehi, K
机构
[1] Mukogawa Womens Univ, Fac Pharmaceut Sci, Nishinomiya, Hyogo 6638179, Japan
[2] Kinnki Univ, Fac Pharmaceut Sci, Higashiosaka, Osaka 5778502, Japan
来源
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 2000年 / 745卷 / 01期
关键词
enantiomer separation; chiral stationary phases; LC; ovoglycoprotein;
D O I
10.1016/S0378-4347(00)00070-0
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The influence of sugar moieties of ovoglycoprotein from chicken egg white (OGCHI) on chiral discrimination of various solutes has been investigated. Partially deglycosylated OGCHI (pd-OGCHI) and completely deglycosylated OGCHI (cd-OGCHI) were obtained by treatments of OGCHI with N-glycosidase, and a mixture of endoglycosidase and N-glycosidase, respectively. The average molecular masses of OGCHI, pd-OGCHI and cd-OGCHI were estimated to be about 30 000, 28 400 and 21 400, respectively, by matrix-assisted laser desorption time-of-flight mass spectrometry. The isoelectric points of OGCHI, pd-OGCHI and cd-OGCHI were in the ranges 4.37-4.51, 4.34-4.44 and 4.17-4.43, respectively, by isoelectric focusing. The OGCHI, pd-OGCHI and cd-OGCHI were bound to aminopropyl-silica gels activated with N,N'-disuccinimidylcarbonate to compare retentive and enantioselective properties of the three columns. It was found that pd-OGCHI showed excellent chiral recognition abilities comparable to OGCHI, and that the retentivity and enantioselectivity of basic solutes tested on the pd-OGCHI column were higher than those on the OGCHI column, while those of acidic solutes tested on the pd-OGCHI column were lower. Further, cd-OGCHI still showed chiral recognition abilities for various solutes tested. These results reveal that the chiral recognition site(s) for OGCHI exists on the protein domain of OGCHI. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:149 / 157
页数:9
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