厌氧条件下扁藻破碎方法及其对氢酶活性的影响研究.pdf

厌氧条件下扁藻破碎方法及其对氢酶活性的影响 1，2 1 1 1 1 1∗ 彦飞 陈兆安 陆洪斌 虞星炬 金美芳 张卫 （1.中国科学院大连化学物理研究所海洋生物产品工程组，116023，大连 2.中国科学院研究生院，100039，北京） 【 】 摘要：亚心型扁藻（Platymonas subcordiformis ）是一种具有光解海水产氢功能的海洋绿藻 1 。为了分离纯化其体内的可溶性氢酶， 本文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 400W, 超声 5 秒，间歇 10 秒，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 2.73mg/ml 。珠磨法和0.2% （v/v ）Triton X-100 破碎细胞的氢酶提取效率较高，分别为 1.44U/g 鲜重细胞和 1.27U/g 鲜重细胞。本实验的最终目的是获得足够量的氢酶进行下一步 纯化，因此珠磨法是最合适的方法。 关键词：微藻制氢；亚心型扁藻；细胞破碎；总蛋白浓度；氢酶活性 中图分类号： 文献标识码：A 文章编号： Fragmentation of Platymonas subcordiformis under anaerobic environment and its effect on hydrogenase activity 1，2 1 1 1 1 1 Yan Fei ,Chen Zhao’an , Lu Hongbin ,Yu Xingju , Jin Meifang , Zhang Wei * (1 Marine Bioproducts Engineering Group, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023 ，China ；2 Graduate School of the Chinese Academy of Sciences, Beijing 100039，China) Abstract ：Platymonas subcordiformis is a kind of marine green algae that can evolve hydrogen by photolysis of seawater. For isolation and purification of dissolvable hydrogenase in alga cells, methods of alga cells fragmentation was studied under anaerobic environment. And the effects of experimental conditions on the hydrogenase activity were compared. The result showed that ultrasonication was a good method for cells fragmentation. The concentration of total protein could arrive 2.73mg/ml, under the condition that the output power, crushing time, interval and times of work respectively were 400W, 5s, 10s and 20.The efficiency of