Cloning of genomic DNA of Rhizopus niveus lipase and expression in the yeast Saccharomyces cerevisiae.
Genomic DNA encoding Lipase I was cloned from Rhizopus niveus strain IFO4759. For expression of this gene in S. cerevisiae, DBY746 was transformed with YEp352PLipS, which had the cloned lipase gene under the control of a PGK promoter. This strain secreted the lipase at a high level (350 U/ml). The strain ND-12B was produced by a mating of DBY746, harboring YEp352PLipS, and NA74-3A, and dissection of asci. This new strain secreted the lipase up to 530 U/ml. Moreover, the lipase was produced most effectively in a medium containing Bacto-yeast extract, soy-peptide, and sucrose.
Kohno
M
M
Central Research Institute, Tsukuba R&D Center, Fuji Oil Co., Ltd., Ibaraki, Japan. kouno@ji.tkc.fujioil.co.jp
Enatsu
M
M
Kugimiya
W
W
D015153:Blotting, Western; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D018274:Electroporation; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D008049:Lipase / Q000737:chemistry / Q000235:genetics*; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012233:Rhizopus / Q000201:enzymology* / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics