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Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger β-galactosidase production

Paper ID Volume ID Publish Year Pages File Format Full-Text
22277 43265 2007 7 PDF Available
Title
Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger β-galactosidase production
Abstract

A flocculent Saccharomyces cerevisiae strain was engineered to stably secrete Aspergillus niger β-galactosidase in a continuous high-cell-density bioreactor. The δ-sequences from the yeast retrotransposon Ty1 were used as target sites for the integration of the β-galactosidase expression cassette. High-copy-number transformants were successfully obtained using the δ-integration system together with the dominant selection antibiotic, G418. The integration of multiple copies was confirmed by genomic Southern blot analysis. Integrants with the highest β-galactosidase levels (approximately eight gene copies) had similar β-galactosidase activities as a recombinant strain carrying the β-galactosidase expression cassette in a YEp-based vector. The β-galactosidase expression cassettes integrated into the yeast genome were stably maintained after eight sequential batch cultures in a nonselective medium. In continuous high-cell-density culture under the same operating conditions, the integrant strain was more stable than the plasmid-carrying strain. To our knowledge, this is the first study of multicopy δ-integrant stability in a continuous bioreactor operating at different dilution rates.

Keywords
genetic stability of delta-integrating systems; continuous high-cell-density culture; Aspergillus niger β-galactosidase production; recombinant; Saccharomyces cerevisiae; yeast flocculation
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Development of stable flocculent Saccharomyces cerevisiae strain for continuous Aspergillus niger β-galactosidase production
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 103, Issue 4, April 2007, Pages 318–324
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