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.

Ämnesord

TEKNIK OCH TEKNOLOGIER  -- Industriell bioteknik (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Industrial Biotechnology (hsv//eng)

Nyckelord

beta-Glucosidase: chemistry

Thermotoga neapolitana: enzymology

Recombinant Fusion Proteins: metabolism

beta-Glucosidase: genetics

beta-Glucosidase: metabolism

Recombinant Fusion Proteins: isolation & purification

Recombinant Fusion Proteins: genetics

Recombinant Fusion Proteins: chemistry

Oligopeptides: metabolism

Oligopeptides: isolation & purification

Oligopeptides: genetics

Oligopeptides: chemistry

Metals: chemistry

Histidine: metabolism

Histidine: isolation & purification

Histidine: genetics

Histidine: chemistry

Escherichia coli: metabolism

Affinity: methods

Chromatography

Escherichia coli: genetics

beta-Glucosidase: isolation & purification

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