Abian Olga, Mateo César, Fernández-Lorente Gloria, Guisán José M, Fernández-Lafuente Roberto
Instituto de Catálisis y Petroleoquímica, CSIC, Universidad Autónoma de Madrid, Cantoblanco 28049 Madrid, Spain.
Biotechnol Prog. 2003 Nov-Dec;19(6):1639-42. doi: 10.1021/bp0340786.
The hydrolysis of penicillin G in the presence of an organic solvent, used with the purpose of extracting it from the culture medium, may greatly simplify the industrial preparation of 6-APA. However, under these conditions, PGA immobilized onto Eupergit displays very low stability (half-life of 5 h in butanone-saturated water) and a significant degree of inhibition by the organic solvent (30%). The negative effect of the organic solvent strongly depended on the type of solvent utilized: water saturated with butanone (around 28% v/v) had a much more pronounced negative effect than that of methylisobutyl ketone (MIBK) (solubility in water was only 2%). These problems were sorted out by using a new penicillin G acylase derivative designed to work in the presence of organic solvents (with each enzyme molecule surrounded by an hydrophilic artificial environment) and a suitable organic solvent (MIBK). Using such solvent, this derivative kept its activity unaltered for 1 week at 32 degrees C. Moreover, the enzyme activity was hardly inhibited by the presence of the organic solvent. In this way, the new enzyme derivative thus prepared enables simplification of the industrial hydrolysis of penicillin G.
为了从培养基中提取青霉素G而在有机溶剂存在的情况下进行青霉素G的水解，可极大地简化6-氨基青霉烷酸（6-APA）的工业制备。然而，在这些条件下，固定在Eupergit上的青霉素G酰化酶（PGA）稳定性非常低（在丁酮饱和水中半衰期为5小时），并且受到有机溶剂的显著抑制（30%）。有机溶剂的负面影响强烈取决于所使用的溶剂类型：丁酮饱和水（约28% v/v）的负面影响比甲基异丁基酮（MIBK）（在水中的溶解度仅为2%）更为明显。通过使用一种设计用于在有机溶剂存在下工作的新型青霉素G酰化酶衍生物（每个酶分子被亲水性人工环境包围）和合适的有机溶剂（MIBK），这些问题得到了解决。使用这种溶剂，该衍生物在32℃下1周内活性保持不变。此外，酶活性几乎不受有机溶剂存在的抑制。通过这种方式，由此制备的新型酶衍生物能够简化青霉素G的工业水解。
