Dialysed activated sludge was used as a substrate to test for enzymes which can hydrolyse activated sludge floc. Two hundred and fifty aerobic and anaerobic microorganisms were tested against activated sludge for the presence of hydrolytic enzymes. These included known genera and organisms obtained by various enrichment procedures. Anaerobic digester mixed liquor was contacted with activated sludge agar under anaerobic conditions. None of the microorganisms present in the digester liquor hydrolysed the floc. The following commercial enzymes were contacted with activated sludge singly, in combination, and sequentially under various physical and chemical conditions: protease, lipase, cellulose, pectinase, phospholipase C, trypsin, and glucuronidase. Although commercial enzymes and various microorganisms reacted with known substrate controls neither the enzymes nor microorganisms employed affected the activated sludge floc. Treatment of activated sludge floc with ethylenediaminetetraacetic acid resulted in gross deflocculation and release of humic substances. The floc particle, probably a combination of polymers bound in a complex manner, is resistant to enzymatic degradation. Several workers have reported on the enzymatic hydrolysis of polymers produced by axenic cultures isolated from activated sludge floc. It is unlikely that the results of their work can be extrapolated to the complex structure of activated sludge floc particles
