Chapter 6-Measuring and reporting enzyme´s immobilization efficiency

PAULA NICOLÁS; CARLA JOSÉ; CARLOS LLERENA SUSTER; VERÓNICA LASSALLE; LAURA BRIAND; MARÍA LUJÁN FERREIRA

Biocatalyst Immobilization

Elsevier

Año: 2022; p. 115 - 147

This chapter presents , discuss and analyzed the approach of the Catalysis Group of PLAPIQUI and the collaborations with CINDECA and CIPROVE in the topic of protein quantification and measurements of immobilized protein in the field of biocatalyst immobilization. It includes information that is not available in articles published in journals and important information to avoid mistakes and erroneousprotein amount measurements.Measuring and reporting enzyme immobilization efficiencies are closely related to the protein quantification methods. Quantification methods detect the protein amount, and the assumption is that there is a correlation between the protein amount and enzyme amount. When using a commercial enzyme, many times isoenzymes are present, as well as other compounds that are not reported in the label. An adequate characterization of the solution or the solid is key, including methods such as SDS-Page, among others. The quantification of proteins should combine at least two methods such as a spectrophotometric method and S-ICP (related to the S content of the protein) or elemental analysis. The Bradford method, even if of widespread use (as well as all spectrophotometric methods using bovine serum albumin as a calibration protein), has several drawbacks in the field of enzymes that have to be considered to avoid mistakes. This chapter analyzes several important aspects related to the quantification of proteins in the enzyme field.Aclaración: Se incluye la proof del capítulo porque se facilitó por parte de Elsevier un link para bajar el libro completo (incluido en el apartado libros editados) pero no los capítulos individuales finales terminados.