A single-step isolation of K99 pili from B-44 strain of Escherichia coli

Abstract

A single-step procedure has been developed to isolate pili from enterotoxigenic Escherichia coli strain B44 of calf origin. Pili, removed from the bacteria by heat shock treatment, were allowed to aggregate at 4°C for 16 h. The precipitated pili, isolated by centrifugation, had (a) typical pili morphology as shown by electron microscopy; (b) ability to bind pig brush border; (c) molecular weight greater than 6 million; and (d) predominance of hydrophobic amino acids. On sodium dodecyl sulfate gel electrophoresis, the subunit pilin migrated as a single polypeptide of molecular weight 17,000.