1. Recombinant Expression of Alliin Lyase from Garlic (Allium sativum) in Bacteria and Yeasts
- Author
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Peter Raspor, Diethard Mattanovich, Andrej Francky, Gerald Striedner, Robert Weik, and Karl Bayer
- Subjects
Protein Folding ,Blotting, Western ,Pharmaceutical Science ,Saccharomyces cerevisiae ,Biology ,medicine.disease_cause ,Pichia ,Inclusion bodies ,Analytical Chemistry ,Microbiology ,Pichia pastoris ,Alliinase ,Drug Discovery ,Escherichia coli ,medicine ,Cloning, Molecular ,Garlic ,Pharmacology ,Plants, Medicinal ,Organic Chemistry ,food and beverages ,Allium sativum ,biology.organism_classification ,Recombinant Proteins ,Yeast ,Carbon-Sulfur Lyases ,Complementary and alternative medicine ,Biochemistry ,Alliin lyase activity ,biology.protein ,Molecular Medicine ,Electrophoresis, Polyacrylamide Gel - Abstract
Recombinant garlic alliin lyase was produced in Escherichia coli, Saccharomyces cerevisiae, and Pichia pastoris. A cDNA clone was obtained from garlic bulbs by PCR and introduced into suitable bacterial and yeast expression vectors. The recombinant alliin lyase forms inclusion bodies in all three host organisms, which are deposited in the cytoplasm. After cell lysis and harvesting by centrifugation, the inclusion bodies were solubilized in Zwittergent 3-14 solution and refolded by stepwise dilution. Specific alliin lyase activity could be recovered by this procedure.
- Published
- 1998
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