Blumberg P M, Strominger J L
Proc Natl Acad Sci U S A. 1972 Dec;69(12):3751-5. doi: 10.1073/pnas.69.12.3751.
An affinity chromatography technique was developed to isolate the five penicillin-binding components present in Bacillus subtilis membranes. The proteins were solubilized by the detergent Nonidet P-40, bound covalently to penicillin-substituted Sepharose, and subsequently eluted from the matrix with neutral hydroxylamine, which cleaves the penicilloyl-enzyme bond. Penicillin binding-component V, the D-alanine carboxypeptidase, makes up 1% of the total membrane protein. A modification of the above procedure enabled this enzyme to be obtained from the membrane in pure form in a single step with 50% overall recovery of enzymatic activity.
开发了一种亲和色谱技术，用于分离枯草芽孢杆菌细胞膜中存在的五种青霉素结合成分。这些蛋白质用去污剂Nonidet P - 40溶解，与青霉素取代的琼脂糖共价结合，随后用中性羟胺从基质上洗脱，中性羟胺可裂解青霉素酰 - 酶键。青霉素结合成分V，即D - 丙氨酸羧肽酶，占总膜蛋白的1%。对上述方法的改进使得该酶能够在一步中以纯形式从细胞膜中获得，酶活性的总回收率为50%。
