摘要
目的:从螺旋藻发酵物中纯化出螺旋藻激酶,并通过质谱分析其蛋白相关信息,为研究其结构和功能提供理论依据。方法:通过切向超滤法、Sephadex G-75凝胶色谱层析、超滤离心管离心法、SDS-PAGE电泳法从螺旋藻发酵物中纯化出螺旋藻激酶,通过基质辅助激光解析电离飞行时间质谱技术分析其蛋白相关信息。结果:获得纯化的单一螺旋藻激酶,其相对分子质量约为45 000,能直接溶解纤维蛋白。基质辅助激光解析电离飞行时间质谱分析表明螺旋藻激酶的氨基酸序列与极大节螺旋藻犬尿氨酸甲酰胺酶有较高相似度,蛋白覆盖率为35%。结论:从螺旋藻发酵物中成功纯化出螺旋藻激酶,并通过质谱获得其蛋白相关信息。
Objective： To purify spirulinakinase from crude fermentative Spirulina and analyze the relevant features of its protein using mass spectrometry for providing theoretical basis for the study of its structure and func- tion. Methods： The spirulinakinase was purified from crude fermentative Spirulina by the methods of cross flow ul- trafiltration, Sephadex G-75 gel filtration chromatography, Milipore uhrafiltration centrifugal tube centrifugation, and SDS-PAGE electrophoresis. The relevant features of spirulinakinase protein were analyzed by MALDI-TOF-TOF mass spectrometry. Results： The spirulinakinase with a molecular weight of 45 000 was purified, which could di- rectly dissolve fibrin. From the results of MALDI-TOF-TOF mass spectrometry, the highly similar sequence of ami- no acids between spirulinakinase and formamidase （Arthrospira maxima） was found, and the protein sequence cov- erage was 35%. Conclusion： Spirulinakinase has been purified successfully from the crude fermentative Spirulina, and the information of its protein has been acquired by mass spectrometric analysis.
出处
《中国新药杂志》
CAS
CSCD
北大核心
2014年第22期2687-2693,共7页
Chinese Journal of New Drugs
基金
广西壮族自治区自然科学基金(2013GXNSFAA019176)
关键词
螺旋藻激酶
分离纯化
质谱分析
纤溶活性
spirulina kinase
separation and purification
mass spectrometric analysis
fibrinolytic activity
