Purification and some properties of protease I having transfer action from Streptomyces griseus var. alcalophilus.

Abstract

Streptomyces griseus var. alcalophilus was selected because it secreted a unique protease (protease I) that catalyzed the transfer reaction forming the hydroxamic acids of various amino acids. Protease I was purified to the electrophoretically homogeneous state and an activity of more than 125-fold that of the culture broth. The molecular weight of the enzymewas estimated to be 25,000 by gel filtration. The enzymewas most active in neutral pHfor the transfer reaction forming phenylalanine hydroxamic acid, although for the hydrolytic reaction with casein as substrate it was most active in alkaline pH. The enzyme was inhibited by diisopropylfluorophosphate.Protease I catalyzed the transfer reaction synthesizing the hydroxamic acids of hydrophobic, acidic, basic, and small aliphatic amino acids such as Phe, Tyr, Leu, Asp, Glu, Arg, Lys, Ala, and Gly. These results indicate that protease I has broad donor specificity. It is also considered that protease I is a unique enzymewith transfer activity, and distinct from the alkalophilic proteinase reported previously [Yamamoto et al., Agric. Biol. Chem., 38, 37 (1974)].Proteases such as trypsin, papain, and achymotrypsin catalyze peptide bond hydrolysis via an acyl-enzyme intermediate. Although the equilibrium of the reaction overwhelmingly favors hydrolysis under physiological conditions, the reaction can be reversed by choosing appropriate conditions so that peptide bonds can be synthesized with a protease as catalyst.

Protease production by Streptomyces thermovulgaris grown on rapemeal‐derived media

“…T h e mol. wts for streptomycete serine proteases range between 22 and 30 kDa (Renko et al 1981(Renko et al , 1989Muro et al 1991) similar to that of the smallest enzyme detected by gel filtration described here. Relatively few streptomycete metalloproteases have been characterized but an enzyme from S. clavulagerus has a mol.…”

Purification and partial characterization of an extracellular serine-proteinase of Streptomyces cyaneus isolated from Brazilian cerrado soil

“…This value was similar to that found for other streptomycetes serine‐proteinases (22 and 30 kDa) ( Renko et al . 1989 ; Muro et al . 1991 ; Yeoman & Edwards 1997).…”

Production and optimization of thermophilic alkaline protease in solid-state fermentation by Streptomyces sp. CN902

“…Currently, commercial proteases are mainly fungal [8] and eubacterial products. Streptomyces species producing protease include S. griseus, S. rimosus and S. thermovulgaris [10][11][12][13]. Protease production has been studied in submerged (SmF) and solid-state fermentation (SSF) [14,15].…”