Purification of native dehydrin from Glycine Max cv., Pisum sativum, and Rosmarinum officinalis by affinity chromatography.
An improved method for the purification of dehydrin from soy (glycine max) is described. Acidic extraction of soy whey was followed by a three step chromatographic process: capture on copper charged Chelating Sepharose Big Beads, intermediate hydrophobic interaction chromatography on Source 15 PHE, and a polishing step on blue Sepharose. The 32-kDa native soy dehydrin was purified to a purity of greater than 98.5% with an overall recovery of 63%. When compared to a previously published purification procedure, recovery, time requirements, and sample preparation steps were improved. The developed method is readily scaleable. Preliminary results show that the process can be used for dehydrins from rosemary (Rosmarinum officinalis) and pea (Pisum sativum).
Herzer
Sibylle
S
Amersham Biosciences, Process Development Department, 800 Centennial Ave., Piscataway, NJ 08855, USA. sibylle.herzer@am.amershambiosciences.com
Kinealy
Kyle
K
Asbury
Reid
R
Beckett
Phil
P
Eriksson
Kjell
K
Moore
Peter
P
D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods*; D002853:Chromatography, Liquid / Q000379:methods; D003300:Copper / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry / Q000379:methods; D018532:Pisum sativum / Q000737:chemistry; D010936:Plant Extracts / Q000737:chemistry; D010940:Plant Proteins / Q000302:isolation & purification*; D010944:Plants / Q000737:chemistry*; D027542:Rosmarinus / Q000737:chemistry; D013025:Glycine max / Q000737:chemistry; D014227:Triazines / Q000737:chemistry
D010936:Plant Extracts; D010940:Plant Proteins; D014227:Triazines; C070556:dehydrin proteins, plant; C012842:Cibacron Blue F 3GA; D003300:Copper