Expression of native and mutant extracellular lipases fromYarrowia lipolytica in Saccharomyces cerevisiae.
Saccharomyces cerevisiae cannot produce extracellular lipase and utilize low-cost lipid substrates. This study aimed to express extracellular lipase from Yarrowia lipolytica in S. cerevisiae, construct recombinant oily substrate consumer strains, and compare the roles of native and mutant Y. lipolytica extracellular lipases in S. cerevisiae. The LIP2 gene of Y. lipolytica DSM3286 and its mutant Y. lipolytica U6 were isolated and cloned by expression vector in S. cerevisiae. New recombinant S. cerevisiae strains FDS100 containing the native LIP2 gene, and FDS101 containing the mutant LIP2 gene were produced 10 and 15 U ml (-1)  extracellular lipase respectively, on a production medium containing olive oil. New recombinant S. cerevisiae strains produce acceptable amount of extracellular lipase in comparison with Y. lipolytica wild-type strains. These strains can utilize olive oil and lipids as low-cost substrates to produce bioethanol, single cell protein and other biotechnologically valuable products. The recombinant S. cerevisiae strain with mutant LIP2 produced lipase with 1.5-fold higher activity. The LIP2 gene of Y. lipolytica was expressed in S. cerevisiae as a heterologous protein without any modifications. Strong components of the Y. lipolytica expression/secretion system could be used for high-level production of recombinant proteins in S. cerevisiae.
Department of Microbiology, Faculty of Science, University of Maragheh, Maragheh, 55181-83111, Iran. f.darvishi@ymail.com
Farshad
F
Darvishi
D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D000431:Ethanol / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D008049:Lipase / Q000235:genetics / Q000378:metabolism*; D050356:Lipid Metabolism; D060847:Metabolic Engineering / Q000379:methods; D000069463:Olive Oil; D010938:Plant Oils / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D025062:Yarrowia / Q000201:enzymology* / Q000235:genetics
D003470:Culture Media; D000069463:Olive Oil; D010938:Plant Oils; D011994:Recombinant Proteins; D000431:Ethanol; D008049:Lipase