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钝顶螺旋藻藻蓝蛋白的化学稳定性

Chemical stabilization of the phycocyanin from cyanobacterium Spirulina platensis.

作者信息

Sun Li, Wang Shumei, Qiao Zhiyong

机构信息

College of Chemical and Biological Sciences and Engineering, Yantai University, Shandong 264005, PR China.

出版信息

J Biotechnol. 2006 Feb 24;121(4):563-9. doi: 10.1016/j.jbiotec.2005.08.017. Epub 2005 Sep 26.

Abstract

Phycocyanin (PC) prepared from a cyanobacterium Spirulina platensis by the DEAE-DE52 cellulose column chromatography that was developed by gradient elution of 50-250 mM phosphate buffer (pH 7.0) was stabilized by its subunits cross-linked covalently with formaldehyde. The single blue band that the chemically stabilized PC showed in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) demonstrated that the stabilized PC still maintained its trimeric aggregate form even after its incubation at 60 degrees C for 3h and at 100 degrees C for 10 min in the denatured buffer containing 5% (w/v) SDS. Moreover, the stabilized PC exhibited similar spectroscopic properties of absorption and fluorescence to those of the native PC, and showed adequate energy coupling with R-phycoerythrin (R-PE) after it was conjugated with R-PE via glutaraldehyde.

摘要

通过用50-250 mM磷酸盐缓冲液(pH 7.0)梯度洗脱开发的DEAE-DE52纤维素柱色谱法从蓝藻钝顶螺旋藻制备的藻蓝蛋白(PC),通过其亚基与甲醛共价交联而稳定。化学稳定的PC在十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)中显示的单一蓝色条带表明,即使在含有5%(w/v)SDS的变性缓冲液中于60℃孵育3小时和在100℃孵育10分钟后,稳定的PC仍保持其三聚体聚集体形式。此外,稳定的PC表现出与天然PC相似的吸收和荧光光谱特性,并且在通过戊二醛与R-藻红蛋白(R-PE)缀合后,与R-PE表现出足够的能量耦合。

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