Genetic modification of the marine-derived yeast Yarrowia lipolytica with high-protein content using a GPI-anchor-fusion expression system.
Yarrowia lipolytica SWJ-1b isolated from the marine fish gut was found to contain 47.6 g of crude protein per 100 g of cell dry weight and had potential use as single cell protein. When the gene encoding enhanced green fluorescent protein (EGFP) was inserted into the surface display plasmid pINA1317-YlCWP110 and expressed in uracil mutant of Y. lipolytica SWJ-1b, the corresponding protein was successfully displayed on the cell surface, and 100% of the yeast cells exhibited the anchored target proteins. We found that yeast cells displaying EGFP were similar to those of Y. lipolytica SWJ-1b. Furthermore, C(18:1) and C(18:3) fatty acids biosynthesis in the marine yeast cells displaying the heterologous EGFP was weakly enhanced compared with that in its wild-type. The results suggest that the marine-derived Y. lipolytica SWJ-1b can be armed with the heterologous protein by the genetic modification and further used as single cell protein.
Wang
Fang
F
Ocean University of China, Qingdao, China.
Yue
Lixi
L
Wang
Lin
L
Madzak
Catherine
C
Li
Jing
J
Wang
Xianghong
X
Chi
Zhenming
Z
D000596:Amino Acids / Q000032:analysis; D003001:Cloning, Molecular / Q000379:methods*; D005227:Fatty Acids / Q000032:analysis; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D014498:Uracil / Q000378:metabolism; D025062:Yarrowia / Q000166:cytology / Q000235:genetics* / Q000378:metabolism
D000596:Amino Acids; D005227:Fatty Acids; D005656:Fungal Proteins; D017261:Glycosylphosphatidylinositols; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014498:Uracil