Purification and characterization of Clostridium perfringens 120-kilodalton collagenase and nucleotide sequence of the corresponding gene.

摘要: Clostridium perfringens type C NCIB 10662 produced various gelatinolytic enzymes with molecular masses ranging from approximately 120 to 80 kDa. A 120-kDa enzyme was present in the largest quantity culture supernatant, and this purified homogeneity on basis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The identified as major collagenase organism, it cleaved typical substrates such azocoll, a synthetic substrate (4-phenylazobenzyloxy-carbonyl-Pro-Leu-Gly-Pro-D-Arg [Pz peptide]), I collagen fibril. In addition, gene (colA) encoding cloned Escherichia coli. Nested deletions were used define coding region colA, sequenced; nucleotide sequence, encodes protein 1,104 amino acids (M(r), 125,966). Furthermore, N-terminal acid sequence which found reading frame, mass mature calculated be 116,339 Da. Analysis primary structure product showed that stretch 86 containing putative signal sequence. Within PLGP, constituting Pz peptide. This may implicated self-processing collagenase. consensus zinc-binding (HEXXH) suggested for vertebrate Zn collagenases is bacterial Vibrio alginolyticus Achromobacter lyticus protease significant homology C. perfringens, suggesting these three are evolutionarily related. Images