Recombinant expression of alliin lyase from garlic (Allium sativum) in bacteria and yeasts.
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.
Weik
R
R
Francky
A
A
Striedner
G
G
Raspor
P
P
Bayer
K
K
Mattanovich
D
D
D015153:Blotting, Western; D013437:Carbon-Sulfur Lyases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005737:Garlic / Q000235:genetics*; D010843:Pichia / Q000235:genetics; D010946:Plants, Medicinal; D017510:Protein Folding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics
D011994:Recombinant Proteins; D013437:Carbon-Sulfur Lyases; C019243:alliin lyase