Abstract
Although the fission yeast Schizosaccharomyces pombe has been used for high-level heterologous protein production, the productivity of secreted human serum transferrin (hTF) has been low, presumably, because the protein harbors twenty disulfide bonds and two Nglycosylation sites. In the present study, we found that overexpression of endogenous putative protein disulfide isomerase (PDI) improved productivity. Whole genome sequence analysis of S. pombe revealed five putative PDI genes and overexpression of two of them, SPAC17H9.14c and SPBC3D6.13c (SpPdi2p or SpPdi3p, respectively), significantly improved the productivity of secreted hTF. GFP-fused SpPdi2p and SpPdi3p were found to localize to the endoplasmic reticulum. Co-overexpression of SpPdi2p or SpPdi3p with hTF coupled with modifications to the growth medium reported in our previous study were able to increase the level of secreted hTF approximately 30-fold relative to conventional conditions.
| Original language | English |
|---|---|
| Pages (from-to) | 1135-1143 |
| Number of pages | 9 |
| Journal | Applied Microbiology and Biotechnology |
| Volume | 86 |
| Issue number | 4 |
| DOIs | |
| Publication status | Published - Apr 1 2010 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Applied Microbiology and Biotechnology
Cite this
Overexpression of protein disulfide isomerases enhances secretion of recombinant human transferrin in Schizosaccharomyces pombe. / Mukaiyama, Hiroyuki; Tohda, Hideki; Takegawa, Kaoru.
In: Applied Microbiology and Biotechnology, Vol. 86, No. 4, 01.04.2010, p. 1135-1143.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Overexpression of protein disulfide isomerases enhances secretion of recombinant human transferrin in Schizosaccharomyces pombe
AU - Mukaiyama, Hiroyuki
AU - Tohda, Hideki
AU - Takegawa, Kaoru
PY - 2010/4/1
Y1 - 2010/4/1
N2 - Although the fission yeast Schizosaccharomyces pombe has been used for high-level heterologous protein production, the productivity of secreted human serum transferrin (hTF) has been low, presumably, because the protein harbors twenty disulfide bonds and two Nglycosylation sites. In the present study, we found that overexpression of endogenous putative protein disulfide isomerase (PDI) improved productivity. Whole genome sequence analysis of S. pombe revealed five putative PDI genes and overexpression of two of them, SPAC17H9.14c and SPBC3D6.13c (SpPdi2p or SpPdi3p, respectively), significantly improved the productivity of secreted hTF. GFP-fused SpPdi2p and SpPdi3p were found to localize to the endoplasmic reticulum. Co-overexpression of SpPdi2p or SpPdi3p with hTF coupled with modifications to the growth medium reported in our previous study were able to increase the level of secreted hTF approximately 30-fold relative to conventional conditions.
AB - Although the fission yeast Schizosaccharomyces pombe has been used for high-level heterologous protein production, the productivity of secreted human serum transferrin (hTF) has been low, presumably, because the protein harbors twenty disulfide bonds and two Nglycosylation sites. In the present study, we found that overexpression of endogenous putative protein disulfide isomerase (PDI) improved productivity. Whole genome sequence analysis of S. pombe revealed five putative PDI genes and overexpression of two of them, SPAC17H9.14c and SPBC3D6.13c (SpPdi2p or SpPdi3p, respectively), significantly improved the productivity of secreted hTF. GFP-fused SpPdi2p and SpPdi3p were found to localize to the endoplasmic reticulum. Co-overexpression of SpPdi2p or SpPdi3p with hTF coupled with modifications to the growth medium reported in our previous study were able to increase the level of secreted hTF approximately 30-fold relative to conventional conditions.
UR - http://www.scopus.com/inward/record.url?scp=77249099943&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77249099943&partnerID=8YFLogxK
U2 - 10.1007/s00253-009-2393-x
DO - 10.1007/s00253-009-2393-x
M3 - Article
C2 - 20013338
AN - SCOPUS:77249099943
VL - 86
SP - 1135
EP - 1143
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 4
ER -