动物组织血管紧张素转化酶的提取纯化及其工艺优化-生物化工专业论文.docxVIP

均表现出较高的活力，其中最适 pH 均为 8.3；而不同温度的条件对 ACE 活 性的影响更为突出，在 42℃时两种 ACE 活性均最强，高于或低于此温度 ACE 活性出现明显下降。通过 ACE 在与不同浓度下的 HHL 底物反应得出 的反应动力学曲线和 Lineweaver-Burk 方程计算得到：猪肺血管紧张素转化 酶 Km 为 1.521mmol·L-1，Vmax 为 17.301nmol·min-1；兔肺血管紧张素转化酶 Km 为 1.058mmol·L-1，Vmax 为 12.107nmol·min-1。 关键词：血管紧张素转化酶 猪肺 兔肺 纯化 性质 SEPARATION AND PURIFICATION OF ANGIOTENSIN CONVERTING ENZYME FROM PIG AND RABBIT LUNG ABSTRACT Angiotensin converting enzyme (ACE) is membrane anchored dipeptidyl carboxypeptidase that plays a very influential role in blood pressure regulation. For further in-depth study the structure and function, biochemical properties, and the binding process between ACE and its inhibitors, we must preparate a large amount of high activity enzyme. In this paper, ACE was separated and purificated from pig lung and rabbit lung. Then we measured the product activity in every step, compared to the primary method in the report before, and present a new optimized process.It made possibility that obtaining a large number of high recovery rate of ACE. The main results are as follows: The material was fresh pig lung from nearby market.After homogenarating with boric acid buffer, the homogenate was prepared in water bath for 1h at 37℃ containing trypsin then centrifugated. 1.6mol·L-1~2.6mol·L-1 ammonium  sulfate fractional precipitated the supernatant. Then the protein was dialyzed. ACE was subjected to sepharose DEAE-Sepharose FF column chromatography while optimizing the loading quantity of sample, the velocity of flow, the concentration and pH of eluent. Then we could get a large amount of high recovery rate of enzyme. The ACE was purified to 39.48-fold with specific activity 0.1303U·mg -1 and activity recovery 59.14%, and ACE molecular weight was 69kDa, which provided plentiful raw materials for the following purification and other work, and gave a good research foundation. Refering to the separation and purification of pig lung ACE, rabbit lung ACE was purificated in the same way. Rabbit