| With the development of the life science, studies of proteins have become increasingly concerned by human, and the separation and purification of proteins has also become an important topic in the scientific community. Among mulriple biomass, waste beer's yeast was used as the research object. Because its surface has abundant functional groups such as hydroxyl, amino, ect, it is easy to cross-linked and modified. Additionally, the beer's yeast has wide sources, cheap and easy to get, which can cut the price of the absorbent. However, due to the poor mechanical strength, low absorption capacity, bad adsorption selectivity, difficult regeneration and other issues, the practical use was limited. Many literatures had reported the method of acidification, alkalization, crosslinking and other simple ways to improve the physical and chemical adsorption properties of beer's yeast, but the results were not satisfactory. According to these problems, we prepared three new adsorbents by cross-linking, and modification, characterized the performance and studied the adsorption behavior of lysozyme, bovine serum albumin, bovine hemoglobin. Specifically as follows:1. The waste beer's yeast was modified with a single triazine reactive dye Cibacron blue F-3GA, the best conditions of synthesis were tested by using orthogonal experiment. Raman spectroscopy, fluorescence electron microscope were used to characterize the modified waste beer's yeast. The effects of initial enzyme concentration, contact time, pH, ionic strength, and temperature on adsorption of lysozyme were studied. The results showed that when the pH was 7.0, the adsorption capacity was 229.1 mg ? g-1. Langmuir, Fruendlich adsorption model were used to study the adsorption dynamic behavior of lysozyme. The adsorption process fitted the Langmuir monolayer adsorption model. Desorption of the lysozyme was achieved using 1.0 mol ? L-1NaCNS, the elution rate was 89%. Modified biomass was used to extract lysozyme from egg white, purification fold was 26.7, and the average recovery of enzyme activity was 73.5%.2. According to the preparation method of the Cibacron blue F-3GA modified biomass, the triazine reactive dye Procion blue MX-R was modified to waste beer's yeast. Raman spectroscopy was used to characterize the modified biomass. Modified biomass was used for lysozyme and bovine serum protein adsorption, the adsorption optimum pH were 7.0 and 5.0, respectively, and the maxmum adsorption capacity were 121.6mg ? g-1 and 237.6 mg ? g-1, respectively. Langmuir, Fruendlich adsorption model were used to study the adsorption dynamic behavior of lysozyme and bovine serum albumin. The adsorption process fitted the Langmuir monolayer adsorption model. Modified biomass was used to extract lysozyme from egg white, purification fold was 22.5, and the average recovery of enzyme activity was 59.0%.3. The Cibacron blue F-3GA modified biomass and Procion blue MX-R modified biomass were used for the adsorption of bovine hemoglobin. The results showed that when the pH was 7.0, the maxmum adsorption capacity of Cibacron blue F-3GA modified biomass and Procion blue MX-R modified biomass for bovine hemoglobin were 329.6mg ? g -1 and 286.8 mg ? g-1, respectively. Infrared spectra, Langmuir, Fruendlich adsorption theory models were used to study the mechanism of bovine hemoglobin, the process did not fit the Langmuir adsorption model. The two kinds of modified biomass for the purification of bovine blood samples, enrichment fold were12.9 and 11.8, respectively.4. Thiourea was modified on the surface of the waste beer yeast, infrared spectroscopy and XPS spectroscopy were used to characterize the modified biomass. Modified biomass was used for the adsorption of bovine hemoglobin, the results showed that the adsorption optimum pH was 7.0, the maximum adsorption capacity was 258.7 mg ? g-1. Langmuir, Fruendlich adsorption model were used to discuss the adsorption behavior of bovine hemoglobin, the process did not meet the Langmuir adsorption model. FTIR spectra were used to discuss the adsorption mechanism, through compared the infrared spectra change of before and after adsorption of bovine hemoglobin. Modified biomass for the purification of bovine blood samples, enrichment fold were 10.0. |
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