Alkaline proteinase isolated from bacillus sp.

An alkaline proteinase K-16 isolated from Bacillus sp. KSM-K16 comprising the following enzymological characteristics:

• (1)-(2) Action/Substrate specificity
• Exhibits degradation activity on casein, hemoglobin, FCS albumin, keratin and elastin at pH 10.0;
• (3) Optimum pH range
• Optimum pH range of 11.0-12.3 when reacted at 40.degree. C. using casein as a substrate;
• (4) pH stability
• Stable in a pH range of 5.0-12.0 in the presence of Ca.sup.2+ when stored in buffer solutions at 25.degree. C. for 2 days;
• (5) Optimum temperature
• Optimum temperature 55.degree. C. when reacted at pH 10.0 using casein as a substrate;
• (6) Heat stability
• Maintains 90% or higher residual activity when treated at 50.degree. C. and at pH 9.5 for 10 minutes;
• (7) Molecular Weight
• 28. 000.+-.1,000 measured by SDS-polyacrylamido gel electrophoresis;
• (8) Isoelectric point 10.5 or higher;
• (9) Effects of metal ions
• Activity inhibited by Hg.sup.2+ and Cu.sup.2+, and heat stability promoted by Ca.sup.+2 ;
• (10) Effects of inhibitors
• Activity not inhibited by ethylenediaminetetraacetic acid, 4-chloromercuribenzoate and antipain, and inhibited by diisopropyl fluorophosphate, phenylmethanesulfonyl fluoride, and chymostatin;
• and
• (11) Effects of surface active agents
• Stable in the presence of sodium linear alkylbenzene sulfonate, sodium polyoxyethylene alkyl sulfate, sodium dodecyl sulfate, sodium .alpha.-olefin sulfonate, sodium alkyl sulfonate, and .alpha.-sulfo-fatty acid ester.