Enhanced iron uptake of Saccharomyces cerevisiae by heterologous expression of a tadpole ferritin gene.
We genetically engineered Saccharomyces cerevisiae to express ferritin, a ubiquitous iron storage protein, with the major heavy-chain subunit of tadpole ferritin. A 450-kDa ferritin complex can store up to 4,500 iron atoms in its central cavity. We cloned the tadpole ferritin heavy-chain gene (TFH) into the yeast shuttle vector YEp352 under the control of a hybrid alcohol dehydrogenase II and glyceraldehyde-3-phosphate dehydrogenase promoter. We confirmed transformation and expression by Northern blot analysis of the recombinant yeast, by Western blot analysis using an antibody against Escherichia coli-expressed TFH, and with Prussian blue staining that indicated that the yeast-expressed tadpole ferritin was assembled into a complex that could bind iron. The recombinant yeast was more iron tolerant in that 95% of transformed cells, but none of the recipient strain cells, could form colonies on plates containing 30 mM ferric citrate. The cell-associated concentration of iron was 500 microg per gram (dry cell weight) of the recombinant yeast but was 210 microg per gram (dry cell weight) in the wild type. These findings indicate that the iron-carrying capacity of yeast is improved by heterologous expression of tadpole ferritin and suggests that this approach may help relieve dietary iron deficiencies in domesticated animals by the use of the engineered yeast as a feed and food supplement.
Institute for Molecular Biology and Genetics, Chonbuk National University, Chonju, Chonbuk, Korea.
Y M
YM
Shin
T H
TH
Kwon
K S
KS
Kim
K S
KS
Chae
D H
DH
Kim
J H
JH
Kim
M S
MS
Yang
D000818:Animals; D015152:Blotting, Northern; D005293:Ferritins / Q000235:genetics* / Q000378:metabolism*; D007501:Iron / Q000378:metabolism*; D007814:Larva / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013054:Spectrophotometry, Atomic; D014170:Transformation, Genetic
D011994:Recombinant Proteins; D005293:Ferritins; D007501:Iron