PURIFICATION OF HEMOCYANIN FROM MARINE GASTROPOD RAPANA THOMASIANA USING AMMONIUM SULFATE PRECIPITATION METHOD

被引：10
作者：
Idakieva, K. [1 ]
Chakarska, I. [1 ]
Ivanova, P. [2 ]
Tchorbanov, A. [3 ]
Dobrovolov, I. [2 ]
Doumanova, L. [3 ]
机构：
[1] Bulgarian Acad Sci, Inst Organ Chem, Sofia, Bulgaria
[2] Inst Fishing Resources, Varna, Bulgaria
[3] Bulgarian Acad Sci, Inst Microbiol, BU-1113 Sofia, Bulgaria
来源：
BIOTECHNOLOGY & BIOTECHNOLOGICAL EQUIPMENT | 2009年 / 23卷 / 03期
关键词：
Rapana thomasiana; gastropod; hemocyanin; ammonium sulfate; AMINO-ACID-SEQUENCE; FUNCTIONAL UNIT; 2; ISOFORMS; DISSOCIATION; RTH1;
D O I：
10.1080/13102818.2009.10817671
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
The aim of this study was to develop an efficient and simple method for isolation of preparative amounts of pure hemocyanin (Hc) from the hemolymph of the marine gastropod Rapana thomasiana. The methods that are usually used, as ultracentrifugation and column chromatography, are expensive and impractical for the large-scale production of Hc. For the ammonium sulfate precipitation method, the concentrated hemolymph was twice precipitated by 38% saturation with crystalline ammonium sulfate. Rapana thomasiana hemocyanin (RtH) was isolated with good yield and high purity, as assessed by gel chromatography, SDS-PAGE, transmission electron microscopy and absorption spectroscopy. This suggests that the ammonium sulfate precipitation is an efficient and useful purification method, suitable for a large-scale RtH preparation.
引用
收藏
页码：1364 / 1367
页数：4
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