Abstract
Mycelium from Aspergillus niger was made permeable to enzyme substrata and cofactors by treatment with the surfactant cetyltrimethylammonium acetate (CTAA). The permeabilization procedure developed permits rapid assaying of a wide range of enzymes and alleviates the necessity crude extracts for measurement of enzyme activities. Enzyme activities in the permeabilized cell assay system (in situ), as compared to that in cell-free extract (in vitro), were similar for most of the enzymes tested, indicating that detection of enzyme activity by either method was comparably efficient and that CTAA has no adverse effect on enzyme activity. An obvious advantage of the permeabilized cell system is a closer approximation of the enzyme environment to the conditions existing in vivo. Disruption of the cells by mechanically stirring yielded an easy method for the determination of the subcellular localisation of a variety of enzymes present in mycelium of A. niger.
Published Version
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