Production of a thermostable alcohol dehydrogenase from Rhodococcus ruber in three different yeast species using the Xplor®2 transformation/expression platform.

Journal of Industrial Microbiology & Biotechnology
Martin GiersbergG Kunze

Abstract

The Xplor®2 transformation/expression platform was employed for comparative assessment of three different yeast species as hosts for synthesis of a thermostable nicotinamide adenine dinucleotide (NAD⁺)-dependent medium-chain alcohol dehydrogenase from Rhodococcus ruber strain 219. Using yeast ribosomal DNA (rDNA) integrative expression cassettes (YRCs) and yeast integrative expression cassettes (YICs) equipped with a selection-marker module and one, two or four expression modules for transformation of auxotrophic Arxula adeninivorans, Hansenula polymorpha, and Saccharomyces cerevisiae strains, quantitative comparison of the yield of recombinant alcohol dehydrogenase RR-ADH6Hp in all three species was carried out. In all cases, the RR-ADH6H gene was expressed under the control of the strong constitutive A. adeninivorans-derived TEF1 promoter, which functions in all yeast species analyzed. Recombinant RR-ADH6Hp accumulated intracellularly in all strains tested. The best yields of active enzyme were obtained from A. adeninivorans, with S. cerevisiae producing intermediate amounts. Although H. polymorpha was the least efficient producer overall, the product obtained was most similar to the enzyme synthesized by R. ruber 219 with re...Continue Reading

References

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Jan 1, 1994·Antonie van Leeuwenhoek·G Kunze, I Kunze
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Dec 20, 2002·Angewandte Chemie·Wolfgang StampferKurt Faber
Jan 31, 2003·Biotechnology and Bioengineering·Wolfgang StampferKurt Faber
Apr 19, 2003·FEMS Yeast Research·Thomas WartmannGotthard Kunze
May 7, 2003·Applied Microbiology and Biotechnology·T WartmannG Kunze
Mar 10, 2004·Biotechnology and Bioengineering·Birgit KosjekWolfgang Kroutil
Jun 4, 2004·Journal of Industrial Microbiology & Biotechnology·Yaroslav TerentievGotthard Kunze
Aug 29, 2006·Journal of Biotechnology·Gerhard SteinbornGotthard Kunze
Jul 28, 2007·FEMS Yeast Research·Gerhard SteinbornGotthard Kunze
May 12, 2011·Applied Microbiology and Biotechnology·Erik BöerGotthard Kunze

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Citations

Nov 7, 2012·Applied Microbiology and Biotechnology·Miguel Álvaro-BenitoGotthard Kunze
Dec 24, 2014·Applied Microbiology and Biotechnology·Marion RauterH- Matthias Vorbrodt
Sep 12, 2013·Journal of Molecular Microbiology and Biotechnology·Anke Trautwein-SchultGotthard Kunze
Jun 11, 2021·Applied Microbiology and Biotechnology·Daniel Ruben Akiola SanyaMahesh B Khot

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