Bonded cells with penicillin acylase activity - prepd. without carrier or autolysis, useful in 6-amino:penicillanic acid prodn.

摘要

Bonded cells with penicillin acylase (I) activity are made by (a) crosslinking the cell contents of a (I)-producing microorganism with a dialdehyde (II); (b) making the crosslinked cells permeable by treating at high temp., and/or with organic solvent and/or with surfactant; (c) reacting the cells in aq. medium with a water-soluble cpd. having >=2 primary and/or sec. amino gps. and (II). Process further comprises (d) sepg. the bonded cells and improving their mechanical strength by partial dehydration at 0 to -30 degrees C with organic water-miscible solvent (opt. mixed with water); and (e) converting to enzymatically-active form by reconstituting with aq. media of pH 7-8, esp. a phosphate buffer. Pref. (II) is glutardialdehyde; the dehydrating solvents are pref. acetone or propanol and stage (c) is either at below mixt. freezing pt. or at 15-25 degrees C, opt. with stirring at 40-60 r.p.m. The bonded cells are used for continuous hydrolysis of natural penicillins to 6-APA which is used to make semi-synthetic penicillins. The 6-APA formed is free of potentially-allergenic protein impurities. The cells can be treated directly after sepn. from the culture liq. and there is no need for a carrier or autolysis. The cells have high specific activity (up to 90% that of native cells); good sedimentation properties and esp. good hydrodynamic properties.