Coexpression of BiP increased antithrombotic hirudin production in recombinant Saccharomyces cerevisiae.
In order to increase a production level of antithrombotic hirudin, BiP was simultaneously expressed in recombinant Saccharomyces cerevisiae strains carrying ten and 15 copies of the hirudin expression cassette integrated in the chromosome. Coexpression of BiP greatly enhanced both cell growth and hirudin production in recombinant S. cerevisiae. Maximum hirudin concentration of 36 mg l(-1) was obtained from batch culture of the ten copy-number transformant concomitantly harboring an episomal copy of the BiP gene under the control of the GAL1 promoter, which is corresponding to a 2.5-fold increase compared with the control strain carrying the genomic BiP gene only. The mean size of the recombinant yeast cells expressing the BiP gene remained at a relatively constant level compared with the control strains of which size increased after the onset of hirudin expression by the GAL10 promoter.
Kim
Myoung-Dong
MD
Department of Food Science and Technology, Research Center for New Bio-Materials in Agriculture, Seoul National University, Suwon 441-744, South Korea.
Han
Ki-Cheol
KC
Kang
Hyun-Ah
HA
Rhee
Sang-Ki
SK
Seo
Jin-Ho
JH
D002455:Cell Division; D002478:Cells, Cultured; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005343:Fibrinolytic Agents; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D005690:Galactose / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D005947:Glucose / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006629:Hirudins / Q000235:genetics / Q000378:metabolism*; D016254:Mutagenesis, Insertional / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D013045:Species Specificity
D005343:Fibrinolytic Agents; D005656:Fungal Proteins; D018840:HSP70 Heat-Shock Proteins; D006629:Hirudins; C060153:KAR2 protein, yeast; D011994:Recombinant Proteins; D005947:Glucose; D005690:Galactose