Kabakov V E, Kliachko N L, Levashov A V
Biokhimiia. 1995 May;60(5):791-7.
Gel filtration under denaturing conditions was used to isolate the alpha- and beta-subunits of penicillin acylase (PA). Refolded subunits were obtained through removing urea by dialysis. Both renatured subunits were catalytically active during hydrolysis of phenylacetic acid p-nitroanilide; this activity decreased after addition of a serine-specific inhibitor--phenylmethanesulfonyl fluoride. The subunits were also active in reversed micelles of Aerosol OT (AOT) in octane, the optimum hydration degree being 11.9 and 17.5 for the light (alpha) and heavy (beta) subunits, respectively. The positions of the maxima were consistent with both theoretically calculated optimum hydration degrees and the earlier reported profile of enzymatic activity for native PA in reversed micelles.
在变性条件下进行凝胶过滤以分离青霉素酰化酶（PA）的α亚基和β亚基。通过透析去除尿素获得重折叠的亚基。两种复性的亚基在对硝基苯乙酸苯酯水解过程中均具有催化活性；加入丝氨酸特异性抑制剂苯甲基磺酰氟后，该活性降低。这些亚基在辛烷中的气溶胶OT（AOT）反胶束中也具有活性，轻（α）亚基和重（β）亚基的最佳水合度分别为11.9和17.5。最大值的位置与理论计算的最佳水合度以及先前报道的天然PA在反胶束中的酶活性曲线一致。
