Sun Li, Wang Shumei, Qiao Zhiyong
College of Chemical and Biological Sciences and Engineering, Yantai University, Shandong 264005, PR China.
J Biotechnol. 2006 Feb 24;121(4):563-9. doi: 10.1016/j.jbiotec.2005.08.017. Epub 2005 Sep 26.
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.
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。