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A "soluble" form of chitin synthetase was separated from a membrane-rich fraction by exposure to the enzyme substrate (uridine diphosphate N-acetyl-D-glucosamine) and activator (N-acetyl-D-glucosamine). The solubilized enzyme catalyzed the synthesis of chitin microfibrils similar, if not identical, to those formed in vivo by the fungus. Cell wall microfibrils were thus abundantly formed in the absence of a living cell or its membranes.