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Application of simulated moving bed chromatography for protein purification: case study with caffeine-N-demethylase B from pseudomonas putida CBB5

AuthorsMoreno-Cid, Juan A. ; Canales, Mario ; Fuente, José de la
Issue Date2013
PublisherStudium Press
CitationAdvances in Biotechnology: 1-13 (2013)
AbstractSimulated moving bed (SMB) is a well‐established technology based on simulating a countercurrent contact between the solid (stationary) phase and liquid phase. It is widely used in separation of sugars, racemic drugs and drug intermediates at commercial scale. SMB has been shown to be more beneficial in terms of productivity such as product concentration, as well reduced use of solvents/buffers, while retaining product purity. We have developed a four‐zone SMB method (Bind, Wash, Elute and Regenerate) using Nickel (HIS‐Select) affinity chromatography, to purify recombinant caffeine‐N‐demethylase B (NdmB). NdmB (Mol.wt: 35 kDa) from Pseudomonas putida CBB5 catalyzes the N‐3‐demethylation of caffeine. This enzyme has been cloned and expressed E. coli as his‐tag protein.
DescriptionChapter 1.-- et al.
Appears in Collections:(IREC) Libros y partes de libros
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