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Quantification of immobilized Candida antarctica lipase B (CALB) using ICP-AES combined with Bradford method.

作者信息

Nicolás Paula, Lassalle Verónica L, Ferreira María L

机构信息

Planta Piloto de Ingeniería Química (PLAPIQUI-UNS-CONICET), Universidad Nacional del Sur, Camino La Carrindanga km 7, 8000, Bahía Blanca, Argentina.

Instituto de Química del Sur (INQUISUR-UNS-CONICET), Avda. Alem 1253, 8000, Bahía Blanca, Argentina.

出版信息

Enzyme Microb Technol. 2017 Feb;97:97-103. doi: 10.1016/j.enzmictec.2016.11.009. Epub 2016 Nov 25.

Abstract

The aim of this manuscript was to study the application of a new method of protein quantification in Candida antarctica lipase B commercial solutions. Error sources associated to the traditional Bradford technique were demonstrated. Eight biocatalysts based on C. antarctica lipase B (CALB) immobilized onto magnetite nanoparticles were used. Magnetite nanoparticles were coated with chitosan (CHIT) and modified with glutaraldehyde (GLUT) and aminopropyltriethoxysilane (APTS). Later, CALB was adsorbed on the modified support. The proposed novel protein quantification method included the determination of sulfur (from protein in CALB solution) by means of Atomic Emission by Inductive Coupling Plasma (AE-ICP). Four different protocols were applied combining AE-ICP and classical Bradford assays, besides Carbon, Hydrogen and Nitrogen (CHN) analysis. The calculated error in protein content using the "classic" Bradford method with bovine serum albumin as standard ranged from 400 to 1200% when protein in CALB solution was quantified. These errors were calculated considering as "true protein content values" the results of the amount of immobilized protein obtained with the improved method. The optimum quantification procedure involved the combination of Bradford method, ICP and CHN analysis.

摘要

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