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High cell-density expression system: A novel method for extracellular production of difficult-to-express proteins

Paper ID Volume ID Publish Year Pages File Format Full-Text
21369 43219 2012 6 PDF Available
Title
High cell-density expression system: A novel method for extracellular production of difficult-to-express proteins
Abstract

Yeast's extracellular expression provides a cost-efficient means of producing industrially useful recombinant proteins. However, depending on the protein to be expressed, the production results in a poor yield, which is occasionally accompanied with loss of the expression plasmid and hence hampered growth of the host in the inducing medium. Here we propose an alternative approach, high cell-density expression, to improve the yield of a certain range of so-called difficult-to-express proteins. In this expression system, recombinant yeast cells resting in stationary phase (OD660 = 3–4) are suspended in a small aliquot of inducing medium to form a high cell-density culture (e.g., OD660 = 15). When applied to the yeast strains harboring Lentinula edodes laccase (Lcc1 or Lcc4) expressing plasmids, the high cell-density system allowed the host cells to synthesize elevated amounts of the laccase which resulted in > 1000- to 6000-fold higher yield than those synthesized in a classical growth-associated manner. The resting cells required aerobic agitation for the maximum production. The production system also worked for other foreign enzymes but not for beta-galactosidase from Aspergillus oryzae or Escherichia coli, likely suggesting an involvement of chaperons that act on a certain range of secretory proteins.

Keywords
Lentinula edodes laccase; Difficult-to-express protein; Extracellular expression; Saccharomyces cerevisiae; High cell-density expression
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High cell-density expression system: A novel method for extracellular production of difficult-to-express proteins
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 113, Issue 2, February 2012, Pages 154–159
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us