CAJ | 학술논문

血管紧张素转化酶(angiotensin converting enzyme, ACE)存在于生物体组织和血液中,并在血压调控方面发挥着重要作用,高效分离纯化ACE对研究其结构和功能具有重要意义。文中将猪肺匀浆液进行酶解处理,优化浆液比、盐析初始蛋白浓度等条件,并经盐析、透析和离子交换层析等步骤,得到高活性的ACE。当猪肺以1：3浆液比匀浆并经胰蛋白酶处理1 h后,其总酶活可增加27.06%；匀浆后初始蛋白浓度为3%时进行盐析,同时将废弃的一级盐析沉淀蛋白再回收处理,盐析过程粗酶纯化倍数为3.94倍,总酶活回收率可达73.73%,较常规处理提高了24.33%以上,经离子交换层析后,ACE酶比活为0.1303 U/mg,总酶活回收率为59.14%。 ACE酶学性质研究表明,ACE的最佳反应温度为42℃,最适pH为8.3。由酶促反应动力学方程计算得到的猪肺ACE的米氏常数Km为1.521 mmol/L,最大反应速率Vmax为17.301 nmol/min。新分离纯化工艺可使ACE收率大大提高。
혈관긴장소전화매(angiotensin converting enzyme, ACE)존재우생물체조직화혈액중,병재혈압조공방면발휘착중요작용,고효분리순화ACE대연구기결구화공능구유중요의의。문중장저폐균장액진행매해처리,우화장액비、염석초시단백농도등조건,병경염석、투석화리자교환층석등보취,득도고활성적ACE。당저폐이1：3장액비균장병경이단백매처리1 h후,기총매활가증가27.06%；균장후초시단백농도위3%시진행염석,동시장폐기적일급염석침정단백재회수처리,염석과정조매순화배수위3.94배,총매활회수솔가체73.73%,교상규처리제고료24.33%이상,경리자교환층석후,ACE매비활위0.1303 U/mg,총매활회수솔위59.14%。 ACE매학성질연구표명,ACE적최가반응온도위42℃,최괄pH위8.3。유매촉반응동역학방정계산득도적저폐ACE적미씨상수Km위1.521 mmol/L,최대반응속솔Vmax위17.301 nmol/min。신분리순화공예가사ACE수솔대대제고。
Angiotensin converting enzyme ( ACE ) is one kind of protease existed in the biological tissue and blood, which plays a very important role in blood pressure regulation. How to purify ACE efficiently is significant for the study of ACE structure and function. Fresh pig lung was homogenized and digested with the trypsin. The influence factors including concentration of homogenization and initial concentration of protein were optimized. High activity ACE was obtained after salting out, di-alysis and ion exchange chromatograph. The enzyme activity was increased by 27. 06% when the pig lung was homogenized at ratio of 1:3 and digested by the trypsin for 1 h. The homogenarate was cen-trifuged and the protein concentration of the supernatant was diluted to 3% before two steps salting-out, while the primary waste deposition was recycled. The ACE was purified to 3. 94 fold and activi-ty recovery 73. 73% which was improved more than 24. 33% in the comparision with traditional treatment. After ion exchange chromatography, the specific activity of ACE was 0. 130 3 U/mg and the activity recovery reached to 59. 14%. The anzymatic characterization of ACE was investigated. The results showed that the optimized catalytic temperature was 42 ℃ and pH of ACE was 8. 3 . The Michaelis constant and the maximum reaction rate of pig lung ACE were 1. 521 mmol/L, 17. 301 nmol/min, respectively. ACE activity recovery rate could be improved significantly by the new purification process.