Proline-specific endopeptidase from Flavobacterium. Purification and properties.

Abstract

Proline-specific endopeptidase (EC 3.4.21.26) was purified 1,400-fold in an overall yield of 10% from cell-free extracts of Flavobacterium meningosepticum by CMcellulose and hydroxyapatite column chromatographies and gel filtration on Sephadex G-150.The purified enzyme was apparently homogeneous as judged by both standard disc gel and sodium dodecyl sulfate (SDS)-gel electrophoresis.The enzyme is an endopeptidase which catalyzes the hydrolysis of Pro-X peptide bonds (and, more slowly, Ala-X bonds), an activity similar to mammalian post-proline cleaving enzyme (Yoshimoto, T.,