Secretion of mouse alpha-amylase from Kluyveromyces lactis.
We constructed two mouse alpha-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 alpha-amylase in YPD medium at a similar level of efficiency. K. lactis transformants secreted glycosylated and non-glycosylated alpha-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 alpha-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.
Tokunaga
M
M
Laboratory of Applied Microbiology, Faculty of Agriculture, Kagoshima University, Japan.
Ishibashi
M
M
Tatsuda
D
D
Tokunaga
H
H
D000818:Animals; D005822:Genetic Vectors; D006031:Glycosylation; D007716:Kluyveromyces / Q000235:genetics*; D051379:Mice; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013213:Starch / Q000378:metabolism; D014170:Transformation, Genetic; D000516:alpha-Amylases / Q000096:biosynthesis*
D011994:Recombinant Proteins; D013213:Starch; D000516:alpha-Amylases