In vitro study of alcohol dehydrogenase and acetaldehyde dehydrogenase encapsulated into human erythrocytes by an electroporation procedure
In vitro study of alcohol dehydrogenase and acetaldehyde dehydrogenase encapsulated into human erythrocytes by an electroporation procedure
复制标题
DOI：
10.1016/s0304-4165(98)00085-3
复制
发表时间：
1998-10-23
影响因子：
3
通讯作者：
Pinilla, M
中科院分区：
文献类型：
--
作者：
Lizano, C;Sanz, S;Pinilla, M
在本研究之前，确定了乙醇脱氢酶(ADH)和/或乙醛脱氢酶(ALDH)电穿孔/重封载人红细胞的最佳条件：300V，1ms脉冲时间，每15min 8次脉冲，37℃重封1h，获得了较高的包封率和载体细胞回收率。细胞体积增加，而血红蛋白含量减少；因此，观察到细胞血红蛋白浓度降低。观察到负荷红细胞的低渗阻力(贯穿渗透脆性曲线)和不变的氧运输能力(由氧平衡曲线给出)。以游离酶溶液为对照，研究了ADH-或ALDH-红细胞(RBC)在4℃和37℃下对时间的稳定性(长达168小时-7天)。两种酶都从载体红细胞释放到培养基中。在相似条件下对载体红细胞的稳定性进行了研究。观察到血液学参数无显著变化。然而，血红蛋白衍生物的形式表现出修饰，ADH-RBC和共包裹的ADH-和ALDH-RBC对乙醇的连续降解随时间的变化(长达70h)表明这些载体RBC作为乙醇完全代谢的试剂。上述性质为在体内使用ADH和ALDH作为载体系统提供了可能性。(C)1998 Elsevier Science B.V.保留所有权利。
The optimal conditions for electroporated/resealed loading of alcohol dehydrogenase (ADH) and/or acetaldehyde dehydrogenase (ALDH) into human erythrocytes were established prior to the study, with the following characteristics: 300 V, 1 ms pulse time, eight pulses every 15 min and 1 h resealing at 37 degrees C. High encapsulation yield and carrier cell recoveries were achieved. Cell volumes increase while hemoglobin contents decrease; in consequence a decrease in cell hemoglobin concentration was observed. A lower hypotonic resistance of loading erythrocytes (throughout osmotic fragility curves) and unaltered oxygen transport capability (as given by oxygen equilibrium curves) were observed. The stability against time (up to 168 h-7 days) of encapsulated individual enzymes, either ADH- or ALDH-red blood cells (RBCs), was studied at 4 degrees C and 37 degrees C, in comparison with that of free enzyme solutions. Both enzymes were released from carrier RBCs to the incubation medium. The stability of carrier RBCs was studied under similar conditions. Non-significant variations in hematological parameters were observed. However, the hemoglobin derivative forms showed modifications, The continuous degradation of ethanol by ADH-RBCs and coencapsulated ADH- and ALDH-RBCs, as a function of time (up to 70 h) suggests the use of these carrier RBCs as agents for complete metabolization of ethanol. The mentioned properties bare the possibility of using ADH and ALDH as carrier systems in in vivo situations. (C) 1998 Elsevier Science B.V. All rights reserved.