Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC

Cerdanyola del Vallès, Spain

Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC

Cerdanyola del Vallès, Spain
SEARCH FILTERS
Time filter
Source Type

Lopez C.,Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC | Rios S.D.,Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC | Lopez-Santin J.,Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC | Caminal G.,Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC | Alvaro G.,Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC
Biochemical Engineering Journal | Year: 2010

Immobilization of PLP-dependent enzymes requires specific studies due to their special cofactor-enzyme bond. These enzymes were immobilized using different methods, and the recombinant serine hydroxymethyltransferase (SHMT) was used as a case study. The immobilization of SHMT on glyoxal-agarose resulted in a high retention yield (70%); however, the reduction step caused an enzymatic activity loss of 80%. The immobilization on Eupergit® C was optimized by considering different ionic strengths, pH and temperatures. SHMT reached 53% retention on this support. Although the enzymatic activity of the derivative decreased by 36% during the treatment with methylamine and washing, it was totally recovered by incubation with the cofactor. SHMT immobilized on Eupergit® C gained thermal stability with respect to the soluble enzyme. Finally, 6-His-tagged SHMT was adsorbed very rapidly on IMAC supports and reached a 98% immobilization yield and enzymatic retention. The capacity of Eupergit® C beads to immobilize PLP-dependent enzymes was corroborated by the immobilization of alanine racemase and aspartate aminotransferase. The final immobilization yields were 85 and 74% respectively, and the derivatives were two- and nine-fold more stable than the soluble enzymes. © 2010 Elsevier B.V. All rights reserved.

Loading Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC collaborators
Loading Unitat de Biocatalisi Aplicada associada al IQAC UAB CSIC collaborators