Journal
JOURNAL OF BIOTECHNOLOGY
Volume 101, Issue 1, Pages 81-87Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/S0168-1656(02)00288-2
Keywords
hirudin; Saccharomyces cerevisiae; BiP; secretion; GAL10 promoter
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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 mg1(-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 GAL 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. (C) 2002 Elsevier Science B.V. All rights reserved.
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