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| Purification and Characterization of Lectin from Gill of Bighead Carp(Aristichthys nobilis) |
| PAN Sai-kun,TANG Jian,GU Xiao-hong,YANG Chao |
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Abstract A fucose-binding lectin from gill of bighead carp(Aristichthys nobilis)was purified and characterized. The purification procedure consisted of extracting soluble proteins in 25 mmol/L Tris-HCl buffer(pH8.5), separating on DEAE-Sepharose FF ion exchange columns followed by gel filtration chromatography on Sephacryl S-200 HR and Superdex 200 10/300 GL. The purified lectin designated GANL showed a single protein band with an apparent molecular mass of 37 kD when submitted to SDS-polyacrylamide gel electrophoresis under reducing conditions. The native molecular mass of GANL determined by gel filtration on a Superdex 200 column was 220 kD and its carbohydrate content was estimated to be 13.4%. Therefore, GANL is a homomultimeric glycoprotein. The purified lectin only agglutinated native erythrocytes from rabbit without the addition of Ca~(2+). Its activity was not inhibited by any of the mono, disaccharides and glycoprotein tested except for fucose. Amino acid composition analysis revealed that the lectin contained high contents of Asp, Glu, Leu, Val, and Lys.GANL was stability with broad pH(5～11)and the optimum pH was 8.0～9.0. The lectin is thermostable with a temperature optima of 50 ℃.
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Received: 25 December 2009
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