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Purification and characterization of a cold‐active protease from psychrotrophic Serratia marcescens AP3801
Author(s) -
Morita Yasutaka,
Kondoh Kenji,
Hasan Quamrul,
Sakaguchi Toshifumi,
Murakami Yuji,
Yokoyama Kenji,
Tamiya Eiichi
Publication year - 1997
Publication title -
journal of the american oil chemists' society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.512
H-Index - 117
eISSN - 1558-9331
pISSN - 0003-021X
DOI - 10.1007/s11746-997-0240-8
Subject(s) - protease , serratia marcescens , metalloproteinase , biochemistry , isoelectric point , proteases , molecular mass , enzyme , peptide sequence , isoelectric focusing , size exclusion chromatography , chemistry , biology , microbiology and biotechnology , escherichia coli , gene
Abstract Protease activity was detected in the culture medium of Serratia marcescens AP3801 grown at 10°C, which was isolated from soil collected from the top of a mountain. The enzyme, designated as CP‐58 protease, was purified to homogeneity from the culture broth by ion exchange and gel filtration chromatographies. The molecular mass of the protease was 58 kDa, and its isoelectric point was close to 6.0. Maximal activity toward azocasein was observed at 40°C and from pH 6.5 to 8.0. The activity was strongly inhibited by 1,10‐phenanthroline, suggesting that the enzyme is a metalloprotease. The N ‐terminal amino acid sequence was Ser‐Leu‐Asn‐Gly‐Lys‐Thr‐Asn‐Gly‐Trp‐Asp‐Ser‐Val‐Asn‐Asp‐Leu‐Leu‐Asn‐Tyr‐His‐Asn‐Arg‐Gly‐Asn (or Asp)‐Gly‐Thr‐Ile‐Asn‐Asn‐Lys‐Pro‐Ser‐Phe‐Asp‐Ile‐Ala. A search through databases for sequence homology aligned CP‐58 protease with metalloprotease. The result of the cleavage pattern of oxidized insulin B‐chain suggests that CP‐58 protease has a broader specificity than other proteases against the peptide substrate.