Theywadgene fromBacillus subtilisencodes a double-zinc aminopeptidase

Authors:
  • LRLarisa Rabinovitch
    Larisa Rabinovitch
  • SSSmadar Shulami
    Smadar Shulami
  • VRVera Reiland
    Vera Reiland
  • GSGil Shoham
    Gil Shoham
  • YSYuval Shoham
    Yuval Shoham
Publisher website
Google Scholar
Abstract
The yet uncharacterized ywad gene from Bacillus subtilis has been cloned and overexpressed in Escherichia coli. The gene product (BSAP) was purified and shown to be an aminopeptidase. The activity of BSAP was optimal at pH 8.4, the enzyme was stable for 20 min at 80 degrees C and its activity was not affected by serine protease and aspartic protease inhibitors, but was completely diminished by the Zn-chelator 1,10-phenanthroline. ZnCl2 was able to restore activity, and the binding stoichiometry of zinc to apo-BSAP indicated two Zn ions per protein molecule. BSAP exhibited high preference toward p-nitroanilide derived Arg, Lys, and Leu synthetic substrates resulting in kcat/Km values of 1-5 x 10(1) s(-1) mM(-1).

This publication has 29 references indexed in Scilit: