Streptomyces griseus aminopeptidase is a calcium-activated zinc metalloprotein. Purification and properties of the enzyme

Identification of the catalytic residues in the double-zinc aminopeptidase from Streptomyces griseus.

The aminopeptidase from Streptomyces griseus (SGAP) has been cloned and expressed in Escherichia coli. By growing the cells in the presence of 1 M sorbitol at 18 degrees C, the protein was obtained in a soluble and active form. The amino acid sequence of the recombinant SGAP contained four amino acids differing from the previously published sequence. Re-sequencing of the native protein indicate...

Different phosphate binding modes of Streptomyces griseus aminopeptidase between crystal and solution states and the status of zinc-bound water.

Phosphate shows a non-competitive inhibition toward a Streptomyces aminopeptidase (sAP) between pH 5.85 (Ki = 0.48 mM) and 9.0 (110 mM), with a pKa of 7.1 likely due to ionization of H2PO4-. This non-competitive inhibition pattern indicates that phosphate binding to sAP in solution is different from that in the crystal structure, where phosphate is bound to the active site Zn(II) ions. Fluoride...

Isolation and characterization of a phage active against streptomyces griseus

The purification and properties of two staphylolytic enzymes from Streptomyces griseus.

1. Two staphylolytic enzymes have been purified from cultures of a soil isolate of Streptomyces griseus. 2. The purified enzymes were shown to be basic proteins of low molecular weight. Each enzyme released N-acetylmuramic acid reducing groups from the cell walls of Staphylococcus aureus. 3. The enzymes lysed whole staphylococci best at higher pH values and lower ionic strengths than when the s...

Purification and characterization of Streptomyces griseus catechol O-methyltransferase.

A soluble (100,000 x g supernatant) methyltransferase catalyzing the transfer of the methyl group of S-adenosyl-L-methionine to catechols was present in cell extracts of Streptomyces griseus. A simple, general, and rapid catechol-based assay method was devised for enzyme purification and characterization. The enzyme was purified 141-fold by precipitation with ammonium sulfate and successive chr...