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Secretion of Mouse α‐Amylase from Kluyveromyces lactis
Author(s) -
TOKUNAGA MASAO,
ISHIBASHI MATSUJIRO,
TATSUDA DAISUKE,
TOKUNAGA HIROKO
Publication year - 1997
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(19970630)13:8<699::aid-yea124>3.0.co;2-n
Subject(s) - kluyveromyces lactis , biology , secretion , kluyveromyces , saccharomyces cerevisiae , shuttle vector , amylase , recombinant dna , plasmid , secretory protein , signal peptide , microbiology and biotechnology , biochemistry , yeast , gene , enzyme , vector (molecular biology)
We constructed two mouse α‐amylase secretion vectors for Kluyveromyces lactis using the well‐characterized signal sequence of the pGKL 128 kDa killer precursor protein. Both PHO5 and PGK expression cassettes from Saccharomyces cerevisiae directed the expression of mouse α‐amylase in YPD medium at a similar level of efficiency. K. lactis transformants secreted glycosylated and non‐glycosylated α‐amylase into the culture medium and both species were enzymatically active. The K. lactis/S. cerevisiae shuttle secretion vector pMI6 was constructed, and K. lactis MD2/1(pMI6) secreted about four‐fold more α‐amylase than S. cerevisiae YNN27 harboring the same plasmid, indicating that K. lactis is an efficient host cell for the secretion and production of recombinant proteins. © 1997 John Wiley & Sons, Ltd.