Abstract
A new anti-inflammatory proteinase designated as kinonase BI has been isolated from the mixture of kinonases produced by Streptomyces kinoluteus. Kinonase BI is purified by gradient column chromatography on DEAE-cellulose and gel filtration on a column of Sephadex G-75. Kinonase BI is considered to be a neutral proteinase of microbial origin. Kinonase BI is of acidic protein nature having pH optimum at 6.5 and optimum temperature at 55° against casein. Kinonase BI is rather stable in a solution of pH 5-9, but labile specially of acidic pH or by heating at 70° for 10 min. The proteolytic activity of kinonase BI is almost lost by addition of 10-3M of a heavy metal ion, Cu2+ or Hg2+, and ethylenediaminetetraacetic acid, but not by addition of ω-chloroacetophenone, p-chloromercuribenzoate diisopropylfluorophosphate and potato trypsin inhibitor at the same concentration. Kinonase BI hydrolyzes bradykinin to arginylprolylprolylglycine, phenyalanylserylproline and phenylalanylarginine. The anti-inflammatory activity on carrageenin-induced edema is observed in kinonase BI at the same level as in kinonase AI or AIII. Kinonase AII containing a trace of kinonase AI is recovered also from the mixture of kinonases. Kinonase AII is presumed to be a kind of leucine amino peptidases. Bradykinin is not hydrolyzed by kinonase AII and the anti-inflammatory activity is not observed in kinonase AII.
