PURIFICATION AND CHARACTERIZATION OF ALKALINE SERINE-PROTEASE FROM AN ALKALOPHILIC STREPTOMYCES SP

SAP, an extracellular alkaline serine protease produced by Streptomyces sp. YSA-130, was purified to homogeneity by CM-Sephadex column chromatography and crystallization. The enzyme was a monomeric protein with a molecular weight of 19,000 as estimated by SDS-PAGE and gel filtration. The amino acid composition and amino-terminal sequence of SAP were similar to those of other bacterial serine proteases, i.e., Streptomyces griseus proteases A and B, Lysobacter enzymogenes alpha-lytic protease and Nocardiopsis dassonvillei subsp. prasina OPC-210 alkaline serine protease NDP-1. The optimum temperature and pH for the enzyme activity were 60-degrees-C and 11.5. The enzyme was stable up 50-degrees-C, and between pHs 4 and 12. The activity was inhibited by Ag+, Hg2+, Co2+, Sodium dodecyl sulfate, N-bromosuccinimide, diisopropyl phosphorofluoridate (DFP), 2,3-butanedione, 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB), iodoacetate, N-ethylmaleimide (NEM), phenylmethanesulfonyl fluoride (PMSF), and phenylglyoxal.