摘要
为了分离纯化亚心型扁藻(Platymonas subcordiformis)体内的可溶性氢酶,研究了在厌氧环境下藻细胞的破碎方法,以及各种破碎条件对氢酶活性的影响.实验结果表明,使用超声法在输出功率为400 W、超声为5 s、间歇为10 s、工作20次的条件下,破碎细胞后溶液中总蛋白含量最高,可达2.73 g/L.珠磨法和体积分数为0.2%的Triton X-100法破碎的鲜细胞的氢酶提取效率较高,分别为1.44 U/g和1.27 U/g,其中使用Triton X-100法破碎后的粗提液所含其他蛋白的种类较少.实验的最终目的是获得足够量的氢酶进行下一步纯化,因此Triton X-100法是最合适的方法.
To isolate and purify dissolvable hydrogenase in Platymonas subcordiffects cells, the scheme for alga cell fragmentation is designed in anaerobic environment, and the effects of experimental conditions on the hydrogenase activity are investigated. By means of ultrasonication, the concentration of total protein approaches to 2.73 g/L under the condition of 400 W output power, 5 s crushing time, 10 s interval for 20 times. The efficiency of hydrogenase isolation gets higher in case of glass beads or 0.2%（volume fraction） Triton X-100 to break up fresh cells to obtain 1.44 U/g and 1.27 U/g respectively, while the other proteins get less in crude extract by Triton X-100 method, which is suggested to gain enough purified hydrogenase.
出处
《西安交通大学学报》
EI
CAS
CSCD
北大核心
2008年第4期497-500,共4页
Journal of Xi'an Jiaotong University
基金
国家重点基础研究发展规划资助项目(2003CB214506)
国家高技术研究发展计划资助项目(2006AA05Z106)
关键词
亚心型扁藻
细胞破碎
总蛋白浓度
氢酶活性
Platyrnonas subcordiformis
cell fragmentation
total protein concentration
hydrogenase activity
