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Expression of human 5‐lipoxygenase cDNA in Escherichia coli
Author(s) -
Noguchi Masato,
Matsumoto Takashi,
Nakamura Motonao,
Noma Masana
Publication year - 1989
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(89)80638-6
Subject(s) - escherichia coli , complementary dna , lipoxygenase , chemistry , biochemistry , microbiology and biotechnology , biology , enzyme , gene
A cDNA for human 5‐lipoxygenase (5LO) was inserted into the vector pKC (constructed from pKK223‐3 by replacing its replication origin with that of pUC18) and expressed in Escherichia coli . The enzyme expressed was purified to homogeneity from the cellular soluble fraction. The purified enzyme showed both 5LO and leukotriene A 4 synthase activities, which were stimulated by Ca 2+ and ATP. Its molecular mass (78 kDa) and NH 2 ‐terminal sequence were identical with those of 5LO purified from human leukocytes. The availability of the expression system will facilitate further studies on its regulation and the reaction mechanism of the enzyme.