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Seratia proteamaculans HY-3 isolated from thenamed arazyme, with an estimated molecular mass of51.5 kDa. The purified enzyme was characterized as havinghigh activities at wide pH and temperature ranges. We furthercharacterized biochemical features of the enzymatic reactionsunder various reaction conditions. The protease eficientlyhydrolyzed a broad range of protein substrates includingalbumin, keratin, and colagen. The dependence of enzymaticactivities on the presence of metal ions such as calcium andzinc indicated that the enzyme is a metaloprotease, togetherthe enzyme was not inhibited by aspartate, cysteine, or serineprotease inhibitors, but strongly inhibited by 1,10-phenanthrolineand EDTA. The araA gene encoding the exoprotease wasisolated as a 5.6 kb BamHI fragment after PCR amplificationusing degenerate primers and subsequent Southern hybridization.The nucleotide sequence revealed that the deduced aminoacid sequences shared extensive similarity with those of theserralysin family of metalloproteases from other enteric bacteria.A gene (inh) encoding a putative protease inhibitor was alsoaraA structural gene.

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