Kabakov V E, Klyachko N L, Levashov A V
Department of Chemical Enzymology, Moscow State University, Russia.
Biochem Mol Biol Int. 1994 Aug;34(1):201-7.
Refolded alpha and beta subunits of penicillin acylase were produced by gel-filtration under denaturing conditions followed by removal of urea through dialysis. Preparations of both renatured subunits hidrolysed specific substrate--phenylacetic acid p-nitroanilide in buffer and in the system of Aerosol OT reversed micelles, the alpha subunit being most catalytically active at Wo = 11.9, while the beta subunit--at Wo = 17.5. There was a good correlation between the position of the found maxima, the theoretically calculated optimal hydration degrees as well as the earlier reported profile of enzymatic activity for native enzyme in reversed micelles.
