Immobilized metal affinity chromatography in open-loop simulated moving bed technology: purification of a heat stable histidine tagged beta-glucosidase.
Open-loop simulated moving bed (SMB) has been used for immobilized metal affinity chromatographic (IMAC) purification of his-tagged beta-glucosidase expressed in E. coli. A simplified approach based on an optimized single column protocol is used to design the open-loop SMB. A set of columns in the SMB represent one step in the chromatographic cycle i.e. there will be one set each of columns for load, wash, elution etc within the SMB. Only the wash and elution are operated with columns in sequence. The beta-glucosidase was purified to almost single band purity with a purification factor of 15 and a recovery of 91%. SMB-performance showed reduced buffer consumption, higher purification fold, a better yield and higher productivity.
Sahoo
Deepti
D
Department of Biotechnology, Lund University, P.O. Box 124, SE-22100 Lund, Sweden.
Andersson
Jonatan
J
Mattiasson
Bo
B
D002846:Chromatography, Affinity / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006639:Histidine / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006358:Hot Temperature; D008670:Metals / Q000737:chemistry*; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D055550:Protein Stability; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D046949:Thermotoga neapolitana / Q000201:enzymology; D001617:beta-Glucosidase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism
C471213:His-His-His-His-His-His; D008670:Metals; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D006639:Histidine; D001617:beta-Glucosidase